Researcher Database

Hisataka Sabe
Faculty of Medicine Physiological Science Biochemistry
Professor

Researcher Profile and Settings

Affiliation

  • Faculty of Medicine Physiological Science Biochemistry

Job Title

  • Professor

Degree

  • (BLANK)
  • (BLANK)

URL

J-Global ID

Research Interests

  • パキシリン   インテグリン   細胞運動   細胞接着   浸潤   シグナル伝達   Arf6   カドヘリン   paxillin   endocytosis   乳癌   p130Cas   AMAP1   E-cadherin   接触阻止   癌細胞   PAG3   endosomal recycling   ARF   AMAP2   単球細胞   癌浸潤   GEP100   マクロファージ   転移   チロシンリン酸化   Git1   PAGs   ArfGAP   分子細胞生物学   Molecular Cell Biology   

Research Areas

  • Life sciences / Medical biochemistry

Academic & Professional Experience

  • 2010 - Today 北海道大学 医学(系)研究科(研究院) 教授
  • 2000 Osaka University Graduate School of Medicine
  • 2000 Visiting Professor, Osaka Uinversity
  • 1999 Kyoto University Graduate School of Biostudies
  • 1999 Visiting Professor, Kyoto University
  • 1998 大阪バイオサイエンス研究所分子生物学部門 研究部長
  • 1998 Head, Department of Molecular Biology, Osaka Bioscience Institute
  • 1995 新技術事業団 さきがけ21 研究員(兼任)
  • 1994 Associate Professor, Institute for Virus Research, Kyoto University
  • 1993 Kyoto University Institute for Virus Research
  • 1993 米国ロックフェラー大学分子腫瘍学研究室 助教授
  • 1993 The Rockefeller University Laboratory of
  • 1990 米国ロックフェラー大学分子腫瘍学研究室 博士研究員
  • 1990 The Rockefeller University Laboratory of
  • 1988 京都大学遺伝子実験施設 助手
  • 1986 京都大学医学部化学教室第一講座 助手
  • 1986 Instructor, Kyoto University School of Medicine
  • 1985 特別研究員(がん)
  • Molecular Oncology Assistant Professor
  • Molecular Oncology Postdoctoral Associate

Education

  •        - 1986  Kyoto University
  •        - 1986  Kyoto University  Graduate School, Division of Medicine
  •        - 1980  Kyoto University  Faculty of Science
  •        - 1980  Kyoto University  Faculty of Science

Association Memberships

  • JAPAN SOCIETY FOR CELL BIOLOGY   日本生化学会   日本癌学会   日本分子生物学会   

Research Activities

Published Papers

  • Otsuka Y, Oikawa T, Yoshino H, Hashimoto S, Handa H, Yamamoto H, Hashimoto A, Sabe H
    Cell communication and signaling : CCS 16 (1) 1  2018/01 [Refereed][Not invited]
  • Oikawa T, Otsuka Y, Onodera Y, Horikawa M, Handa H, Hashimoto S, Suzuki Y, Sabe H
    Scientific reports 8 (1) 1595  2018/01 [Refereed][Not invited]
  • Yuichi Mazaki, Yasuhito Onodera, Tsunehito Higashi, Takahiro Horinouchi, Tsukasa Oikawa, Hisataka Sabe
    CELL COMMUNICATION AND SIGNALING 15 (1) 36  1478-811X 2017/10 [Refereed][Not invited]
     
    Background: The small GTPase ARF1 mediates membrane trafficking mostly from the Golgi, and is essential for the G protein-coupled receptor (GPCR)-mediated chemotaxis of neutrophils. In this process, ARF1 is activated by the guanine nucleotide exchanger GBF1, and is inactivated by the GTPase-activating protein GIT2. Neutrophils generate the G beta gamma.-PAK1-aPIX-GIT2 linear complex during GPCR-induced chemotaxis, in which aPIX activates RAC1/CDC42, which then employs PAK1. However, it has remained unclear as to why GIT2 is included in this complex. Results: We investigated the association between ARF1 and RAC1/CDC42 during the fMLP-stimulated chemotaxis of HL60 cells. We found that the silencing of GBF1 significantly impaired the recruitment of RAC1 to the leading edges, but not PAK1, aPIX, RAC2, or CDC42. A significant population of RAC1 colocalized with ARF1 at the leading edges in stimulated cells, whereas fMLP activated both ARF1 and ARF5. Consistently, the silencing of ARF1, but not ARF5, impaired the recruitment of RAC1, whereas the silencing of RAC1 did not affect the recruitment of ARF1 to the leading edges. Conclusions: Our results indicated that the activation of ARF1 triggers the plasma membrane recruitment of RAC1 in GPCR-mediated chemotaxis, which is essential for cortical actin remodeling. Thus, membrane remodeling at the leading edges appears to precede actin remodeling in chemotaxis. Together with the fact that GIT2, which inactivates ARF1, is an integral component of the machinery activating RAC1, we proposed a model in which the ARF1-RAC1 linkage enables the regulation of ARF1 by repetitive on/off cycles during GPCR-mediated neutrophil chemotaxis.
  • Tatsuaki Daimon, Takeo Kosaka, Eiji Kikuchi, Shuji Mikami, Yasumasa Miyazaki, Ari Hashimoto, Shigeru Hashimoto, Ryuichi Mizuno, Akira Miyajima, Yasunori Okada, Hisataka Sabe, Mototsugu Oya
    UROLOGIC ONCOLOGY-SEMINARS AND ORIGINAL INVESTIGATIONS 35 (9) 543.e17 - 543.e24 1078-1439 2017/09 [Refereed][Not invited]
     
    Objectives: The erythrocyte protein band 4.1-like5 (EPB4.1L5) regulates E-cadherin in cancer invasion and metastasis inducing epithelial-to-mesenchymal transition. This study aimed to investigate the biological significance of EPB4.1L5 in upper urinary tract urothelial carcinoma (UTUC). Methods: Retrospective analysis of the clinical records of 165 patients with UTUC (Ta-4NOMO) subjected to radical nephroureterectomy and immunohistochemical examination of EPB4.1L5 expression in those tissues. Results: The median follow-up period was 62.2 months (interquartile range = 77.0). The score of EPB4.1L5 significantly correlated with tumor grade, pathological T stage, and lymphovascular invasion (all P < 0.001). The 5-year Kaplan-Meier recurrence-free survival and cancer-specific survival rates were 54.1% and 59.5% in patients with high EPB4.1L5 expression, compared with 81.6% and 87.2%, (all P < 0.001) in their counterparts. Multivariate analyses revealed that high expression of EPB4.1L5 was one of the independent prognostic factors for tumor recurrence (P = 0.022, HR = 2.40) and cancer-specific survival (P = 0.015, HR = 2.94). Conclusion: High EPB4.1L5 expression was related to worse clinical outcome in patients with UTUC. These results indicated that EPB4.1L5 could provide prognostic information in patients with UTUC regarding epithelial-to-mesenchymal transition. (C) 2017 Elsevier Inc. All rights reserved.
  • Nobuyuki Tanaka, Takeo Kosaka, Yasumasa Miyazaki, Shuji Mikami, Naoya Niwa, Yutaro Otsuka, Yoji Andrew Minamishima, Ryuichi Mizuno, Eiji Kikuchi, Akira Miyajima, Hisataka Sabe, Yasunori Okada, Per Uhlén, Makoto Suematsu, Mototsugu Oya
    JCI insight 1 (18) e83654  2016/11/03 [Refereed][Not invited]
     
    To identify the molecules involved in epithelial to mesenchymal transition (EMT) in urothelial carcinoma (UC) after acquisition of platinum resistance, here we examined the changes in global gene expression before and after platinum treatment. Four invasive UC cell lines, T24, 5637, and their corresponding sublines T24PR and 5637PR with acquired platinum resistance, were assessed by microarray, and the ubiquitin E3 ligase FBXO32 was newly identified as a negative regulator of EMT in UC tumors after acquisition of platinum resistance. In vitro and in vivo studies showed an intimate relationship between FBXO32 expression and EMT, demonstrating that FBXO32 dysregulation in T24PR cells results in elevated expression of the mesenchymal molecules SNAIL and vimentin and decreased expression of the epithelial molecule E-cadherin. The association between FBXO32 expression and EMT was further validated using clinical samples. Knockdown of MyoD expression, a specific target of FBXO32 polyubiquitination, revealed upregulation of E-cadherin expression and downregulation of SNAIL and vimentin expression in T24PR cells. Comparative genomic hybridization array analysis demonstrated loss of heterozygosity at 8q24.13 in T24PR cells, which harbors FBXO32. Our findings suggest the importance of the association between EMT and ubiquitin-proteasome regulation when tumors develop acquired platinum resistance.
  • Yutaro Otsuka, Hiroki Sato, Tsukasa Oikawa, Yasuhito Onodera, Jin-Min Nam, Ari Hashimoto, Kiyoshi Fukunaga, Kanako C. Hatanaka, Yutaka Hatanaka, Yoshihiro Matsuno, Satoshi Fukuda, Hisataka Sabe
    CELL COMMUNICATION AND SIGNALING 14 (1) 28  1478-811X 2016/11 [Refereed][Not invited]
     
    Background: Squamous cell carcinoma of the tongue (tongue SCC) is a major subtype of head and neck squamous cell carcinoma (HNSCC), which is an intractable cancer under current therapeutics. ARF6 and its effector AMAP1 are often overexpressed in different types of cancers, such as breast cancer and renal cancer, and in these cancers, AMAP1 binds to EPB41L5 to promote invasion, metastasis, and drug resistance. EPB41L5 is a mesenchymal-specific protein, normally induced during epithelial-mesenchymal transition (EMT) to promote focal adhesion dynamics. Similarly to breast cancer and renal cancer, the acquisition of mesenchymal phenotypes is the key process that drives the malignancy of HNSCC. We previously showed that the overexpression of AMAP1 in tongue SCC is statistically correlated with the poor outcome of patients. In this study, we examined whether tongue SCC also expresses EPB41L5 at high levels. Results: Immunohistochemical staining of clinical specimens of tongue SCC demonstrated that high expression levels of EPB41L5 statistically correlate with poor disease-free survival and poor overall survival rates of patients. The tongue SCC cell line SCC-9, which overexpress Arf6 and AMAP1, also expressed EPB41L5 at high levels to promote invasiveness, whereas the weakly invasive SCC-25 cells did not express EPB41L5 at notable levels. Among the different EMT-associated transcriptional factors, ZEB1 was previously found to be most crucial in inducing EPB41L5 in breast cancer and renal cancer. In contrast, expression levels of ZEB1 did not correlate with the expression levels of EPB41L5 in tongue SCC, whereas KLF8 and FOXO3 levels showed positive correlations with EPB41L5 levels. Moreover, silencing of EPB41L5 only marginally improved the drug resistance of SCC-9 cells, even when coupled with ionizing radiation. Conclusion: Our results indicate that activation of the cancer mesenchymal program in tongue SCC, which leads to EPB41L5 expression, closely correlates with the poor prognosis of patients. However, ZEB1 was not the major inducer of EPB41L5 in tongue SCC, unlike in breast cancer and renal cancer. Thus, processes that trigger the mesenchymal program of tongue SCC, which drives their malignancies, seem to be substantially different from those of other cancers.
  • Handa H, Hashimoto A, Hashimoto S, Sabe H
    Small GTPases 1 - 7 2154-1248 2016/10 [Refereed][Not invited]
  • Sabe H, Hashimoto A, Hashimoto S, Oikawa T
    Molecular & cellular oncology 3 (4) e1185564  2016/07 [Refereed][Not invited]
  • Ari Hashimoto, Tsukasa Oikawa, Shigeru Hashimoto, Hirokazu Sugino, Ayumu Yoshikawa, Yutaro Otsuka, Haruka Handa, Yasuhito Onodera, Jin-Min Nam, Chitose Oneyama, Masato Okada, Mitsunori Fukuda, Hisataka Sabe
    JOURNAL OF CELL BIOLOGY 213 (1) 81 - 95 0021-9525 2016/04 [Refereed][Not invited]
     
    Drug resistance, metastasis, and a mesenchymal transcriptional program are central features of aggressive breast tumors. The GTPase Arf6, often overexpressed in tumors, is critical to promote epithelial mesenchymal transition and invasiveness. The metabolic mevalonate pathway (MVP) is associated with tumor invasiveness and known to prenylate proteins, but which prenylated proteins are critical for MVP-driven cancers is unknown. We show here that MVP requires the Arf6-dependent mesenchymal program. The MVP enzyme geranylgeranyl transferase II (GGT-II) and its substrate Rab11b are critical for Arf6 trafficking to the plasma membrane, where it is activated by receptor tyrosine kinases. Consistently, mutant p53, which is known to support tumorigenesis via MVP, promotes Arf6 activation via GGT-II and Rab11b. Inhibition of MVP and GGT-II blocked invasion and metastasis and reduced cancer cell resistance against chemotherapy agents, but only in cells overexpressing Arf6 and components of the mesenchymal program. Overexpression of Arf6 and mesenchymal proteins as well as enhanced MVP activity correlated with poor patient survival. These results provide insights into the molecular basis of MVP-driven malignancy.
  • Shigeru Hashimoto, Shuji Mikami, Hirokazu Sugino, Ayumu Yoshikawa, Ari Hashimoto, Yasuhito Onodera, Shotaro Furukawa, Haruka Handa, Tsukasa Oikawa, Yasunori Okada, Mototsugu Oya, Hisataka Sabe
    NATURE COMMUNICATIONS 7 10656  2041-1723 2016/02 [Refereed][Not invited]
     
    Acquisition of mesenchymal properties by cancer cells is critical for their malignant behaviour, but regulators of the mesenchymal molecular machinery and how it is activated remain elusive. Here we show that clear cell renal cell carcinomas (ccRCCs) frequently utilize the Arf6-based mesenchymal pathway to promote invasion and metastasis, similar to breast cancers. In breast cancer cells, ligand-activated receptor tyrosine kinases employ GEP100 to activate Arf6, which then recruits AMAP1; and AMAP1 then binds to the mesenchymal-specific protein EPB41L5, which promotes epithelial-mesenchymal transition and focal adhesion dynamics. In renal cancer cells, lysophosphatidic acid (LPA) activates Arf6 via its G-protein-coupled receptors, in which GTP-G alpha 12 binds to EFA6. The Arf6-based pathway may also contribute to drug resistance. Our results identify a specific mesenchymal molecular machinery of primary ccRCCs, which is triggered by a product of autotaxin and it is associated with poor outcome of patients.
  • Dat Nguyen Tien, Masako Kishihata, Ayumu Yoshikawa, Ari Hashimoto, Hisataka Sabe, Eiichiro Nishi, Kaeko Kamei, Hidenori Arai, Toru Kita, Takeshi Kimura, Masayuki Yokode, Noboru Ashida
    SCIENTIFIC REPORTS 4 5094  2045-2322 2014/05 [Refereed][Not invited]
     
    NF-kappa B is a major transcriptional factor regulating many cellular functions including inflammation; therefore, its appropriate control is of high importance. The detailed mechanism of its activation has been well characterized, but that of negative regulation is poorly understood. In this study, we showed AMAP1, an Arf-GTPase activating protein, as a negative feedback regulator for NF-kappa B by binding with IKK beta, an essential kinase in NF-kappa B signaling. Proteomics analysis identified AMAP1 as a binding protein with IKK beta. Overexpression of AMAP1 suppressed NF-kappa B activity by interfering the binding of IKK beta and NEMO, and deletion of AMAP1 augmented NF-kappa B activity. The activation of NF-kappa B induced translocation of AMAP1 to cytoplasm from cell membrane and nucleus, which resulted in augmented interaction of AMAP1 and IKK beta. These results demonstrated a novel role of AMAP1 as a negative feedback regulator of NF-kappa B, and presented it as a possible target for anti-inflammatory treatments.
  • Erythrocyte protein bond 4.1-like5 expression in patients with upper urinary tract urothelial carcinoma correlates with tumor recurrence
    Tatsuaki Daimon, Takeo Kosaka, Shuji Mikami, Yasumasa Miyazaki, Ryuichi Mizuno, Eiji Kikuchi, Akira Miyajima, Ken Nakagawa, Hisataka Sabe, Mototsugu Oya
    JOURNAL OF CLINICAL ONCOLOGY 32 (15) 0732-183X 2014/05 [Refereed][Not invited]
  • Hiroki Sato, Kanako C. Hatanaka, Yutaka Hatanaka, Hiromitsu Hatakeyama, Ari Hashimoto, Yoshihiro Matsuno, Satoshi Fukuda, Hisataka Sabe
    CELL COMMUNICATION AND SIGNALING 12 17  1478-811X 2014/03 [Refereed][Not invited]
     
    Background: Despite recent advances in cancer therapeutics in general, the survival of patients with head and neck squamous cell carcinomas (HNSCCs) has not improved substantially over the past few decades. HNSCC cells often exhibit invasive and metastatic phenotypes, and expression of epidermal growth factor receptor (EGFR) and cortactin has been highly implicated in the development of malignancy in HNSCCs. We have shown previously that an Arf6 pathway, in which Arf6 is activated by GEP100 and employs AMAP1 (also called DDEF1 or ASAP1) as its downstream effector, is pivotal for the invasion and metastasis of different breast cancer cells. This pathway is activated by receptor tyrosine kinases, including EGFR; and moreover, AMAP1 physically associates with cortactin, in which inhibition of this binding effectively blocks invasion and metastasis. We here investigated whether the expression of Arf6 pathway components correlates with the poor prognosis of HNSCC patients. We have shown previously that AMAP1 protein levels are not correlated with its mRNA levels, and hence we here employed immunohistochemical staining of HNSCC clinical specimens to investigate AMAP1 protein levels. Results: We found that high levels of AMAP1 protein expression on its own, as well as its co-overexpression with EGFR statistically correlates with poor disease-free survival and poor overall survival, while high levels of cortactin expression or its co-expression with EGFR did not. Conclusion: Our identification of predictive biomarkers, together with our previous findings on the coherent signaling pathway that these biomarkers ultimately generate should be powerful information for the further development of HNSCC therapeutics.
  • Yasuhito Onodera, Jin-Min Nam, Hisataka Sabe
    PHARMACOLOGY & THERAPEUTICS 140 (1) 1 - 9 0163-7258 2013/10 [Refereed][Not invited]
     
    Integrins are heterodimeric cell surface receptors, which principally mediate the interaction between cells and their extracellular microenvironments. Because of their pivotal roles in cancer proliferation, survival, invasion and metastasis, integrins have been recognized as promising targets for cancer treatment. As is the case with other receptors, the localization of integrins on the cell surface has provided opportunities to block their functions by various inhibitory monoclonal antibodies. A number of small molecule agents blocking integrin-ligand binding have also been established, and some such agents are currently on the market or in clinical trials for some diseases including cancer. This review exclusively focuses on another strategy for cancer therapy, which comes from the obligate localization of integrins on the cell surface; targeting the intracellular trafficking of integrins. A number of studies have shown the essential roles of integrin trafficking in hallmarks of cancer, such as activation of oncogenic signaling pathways as well as acquisition of invasiveness. Recent findings have shown that increased integrin recycling activity is associated with some types of gain-of-function mutations of p53, a common feature of diverse types of cancers, which also indicates that targeting integrin recycling could be widely applicable and effective against many cancers. We also discuss possible therapeutic contexts where integrin trafficking can be effectively targeted, and what molecular interfaces may hopefully be druggable. (C) 2013 Elsevier Inc. All rights reserved.
  • Rumiko Kinoshita, Jin-Min Nam, Yoichi M. Ito, Kanako C. Hatanaka, Ari Hashimoto, Haruka Handa, Yutaro Otsuka, Shigeru Hashimoto, Yasuhito Onodera, Mitsuchika Hosoda, Shunsuke Onodera, Shinichi Shimizu, Shinya Tanaka, Hiroki Shirato, Mishie Tanino, Hisataka Sabe
    PLOS ONE 8 (10) e76791  1932-6203 2013/10 [Refereed][Not invited]
     
    A major problem of current cancer research and therapy is prediction of tumor recurrence after initial treatment, rather than the simple biological characterization of the malignancy and proliferative properties of tumors. Breast conservation therapy (BCT) is a well-approved, standard treatment for patients with early stages of breast cancer, which consists of lumpectomy and whole-breast irradiation. In spite of extensive studies, only 'age' and 'Ki-67 positivity' have been identified to be well correlated with local recurrence after BCT. An Arf6 pathway, activated by GEP100 under receptor tyrosine kinases (RTKs) and employs AMAP1 as its effector, is crucial for invasion and metastasis of some breast cancer cells. This pathway activates beta 1 integrins and perturbs E-cadherin-based adhesions, hence appears to be integral for epithelial-mesenchymal transdifferentiation (EMT). We here show that expression of the Arf6 pathway components statistically correlates with rapid local recurrence after BCT. We retrospectively analyzed four hundred seventy-nine patients who received BCT in Hokkaido University Hospital, and found 20 patients had local recurrence. We then analyzed pathological samples of patients who experienced local recurrence by use of Kaplan-Meier analysis, Stepwise regression analysis and the t-test, coupled with immunostaining, and found that co-overexpression of GEP100 and AMAP1 correlates with rapidity of the local recurrence. Their margin-status, node-positivity, and estrogen receptor (ER)-or progesterone receptor (PgR)positivity did not correlated with the rapidity. This study is the first to show that expression of a certain set of proteins correlates with the rapidity of local recurrence. Our results are useful not only for prediction, but highlight the possibility of developing novel strategies to block local recurrence. We also discuss why mRNAs encoding these proteins have not been identified to correlate with local recurrence by previous conventional gene expression profiling analyses.
  • Nam JM, Ahmed KM, Costes S, Zhang H, Onodera Y, Olshen AB, Hatanaka KC, Kinoshita R, Ishikawa M, Sabe H, Shirato H, Park CC
    Breast cancer research : BCR 4 15 (4) R60  1465-5411 2013 [Refereed][Not invited]
  • Yuichi Mazaki, Yasuharu Nishimura, Hisataka Sabe
    MOLECULAR BIOLOGY OF THE CELL 13 23 (13) 2457 - 2467 1059-1524 2012/07 [Refereed][Not invited]
     
    Most chemoattractants for neutrophils bind to the G alpha(i) family of heterotrimeric G protein-coupled receptors (GPCRs) and release G beta gamma subunits to activate chemotaxis and superoxide production. GIT2, a GTPase-activating protein for Arf1, forms a complex with G beta gamma and is integral for directional sensing and suppression of superoxide production. Here we show that GBF1, a guanine nucleotide exchanging factor for Arf-GTPases, is primarily responsible for Arf1 activation upon GPCR stimulation and is important for neutrophil chemotaxis and superoxide production. We find that GBF1 bears a novel module, namely binding to products of phosphatidyl inositol 3-kinase (PI3K), which binds to products of PI3K gamma. Through this binding, GBF1 is translocated from the Golgi to the leading edge upon GPCR stimulation to activate Arf1 and recruit p22phox and GIT2 to the leading edge. Moreover, GBF1-mediated Arf1 activation is necessary to unify cell polarity during chemotaxis. Our results identify a novel mechanism that links PI3K gamma activity with chemotaxis and superoxide production in GPCR signaling.
  • Jian Li, Andrew W. Malaby, Michael Famulok, Hisataka Sabe, David G. Lambright, Victor W. Hsu
    DEVELOPMENTAL CELL 22 (6) 1286 - 1298 1534-5807 2012/06 [Refereed][Not invited]
     
    The glucose transporter type 4 (glut4) is critical for metabolic homeostasis. Insulin regulates glut4 by modulating its expression on the cell surface. This regulation is mainly achieved by targeting the endocytic recycling of glut4. We identify general receptor for 3-phosphoinositides 1 (Grp1) as a guanine nucleotide exchange factor for ADP-ribosylation factor 6 (ARF6) that promotes glut4 vesicle formation. Grp1 also promotes the later steps of glut4 recycling through ARF6. Insulin signaling regulates Grp1 through phosphorylation by Akt. We also find that mutations that mimic constitutive phosphorylation of Grp1 can bypass upstream insulin signaling to induce glut4 recycling. Thus, we have uncovered a major mechanism by which insulin regulates glut4 recycling. Our findings also reveal the complexity by which a single small GTPase in vesicular transport can coordinate its multiple steps to accomplish a round of transport.
  • Yasuhito Onodera, Jin-Min Nam, Ari Hashimoto, Jim C. Norman, Hiroki Shirato, Shigeru Hashimoto, Hisataka Sabe
    JOURNAL OF CELL BIOLOGY 7 197 (7) 983 - 996 0021-9525 2012/06 [Refereed][Not invited]
     
    Epidermal growth factor receptor (EGFR) signaling is one of the crucial factors in breast cancer malignancy. Breast cancer cells often overexpress Arf6 and its effector, AMAP1/ASAP1/DDEF1; in these cells, EGFR signaling may activate the Arf6 pathway to induce invasion and metastasis. Active recycling of some integrins is crucial for invasion and metastasis. Here, we show that the Arf6-AMAP1 pathway links to the machinery that recycles beta 1 integrins, such as alpha 3 beta 1, to promote cell invasion upon EGFR stimulation. We found that AMAP1 had the ability to bind directly to PRKD2 and hence to make a complex with the cytoplasmic tail of the beta 1 subunit. Moreover, GTP-Rab5c also bound to AMAP1, and activation of Rab5c by EGFR signaling was necessary to promote the intracellular association of AMAP1 and PRKD2. Our results suggest a novel mechanism by which EGFR signaling promotes the invasiveness of some breast cancer cells via integrin recycling.
  • Yuki Wakayama, Koichi Miura, Hisataka Sabe, Naoki Mochizuki
    JOURNAL OF BIOLOGICAL CHEMISTRY 286 (51) 44243 - 44253 0021-9258 2011/12 [Refereed][Not invited]
     
    The epithelial cells exhibit either a columnar or a flat shape dependent on extracellular stimuli or the cell-cell adhesion. Membrane-anchored ephrinA stimulates EphA receptor tyrosine kinases as a ligand in a cell-cell contact-dependent manner. The mechanism through which ephrinA1/EphA2 signal regulates the cell morphology remains elusive. We demonstrate here that ephrinA1/EphA2 signal induces compaction and enhanced polarization (columnar change) of Madin-Darby canine kidney epithelial cells by regulating Ezrin, a linker that connects plasma membrane and actin cytoskeleton. Activation of EphA2 resulted in RhoA inactivation through p190RhoGAP-A and subsequent dephosphorylation of Ezrin on Thr-567 phosphorylated by Rho kinase. Consistently, the cells expressing an active mutant of Ezrin in which Thr-567 was replaced with Asp did not change their shape in response to ephrinA1. Furthermore, depletion of Ezrin led to compaction and enhanced polarization without ephrinA1 stimulation, suggesting the role for active Ezrin in keeping the flat cell shape. Ezrin localized to apical domain irrespective of ephrinA1 stimulation, whereas phosphorylated Ezrin on the apical domain was reduced by ephrinA1 stimulation. Collectively, ephrinA1/EphA2 signal negatively regulates Ezrin and promotes the alteration of cell shape, from flat to columnar shape.
  • Toshi Menju, Shigeru Hashimoto, Ari Hashimoto, Yutaro Otsuka, Haruka Handa, Eiji Ogawa, Yoshinobu Toda, Hiromi Wada, Hiroshi Date, Hisataka Sabe
    PLOS ONE 6 (9) e25301  1932-6203 2011/09 [Refereed][Not invited]
     
    Overexpression of Her2/ErbB2/Neu in cancer is often correlated with recurrent distant metastasis, although the mechanism still remains largely elusive. We have previously shown that EGFR, when tyrosine-phosphorylated, binds to GEP100/BRAG2 to activate Arf6, which induces cancer invasion and metastasis. We now show that overexpressed Her2 in lung adenocarcinoma cells also employs GEP100. Like EGFR-GEP100 binding, this association is primarily mediated by the pleckstrin homology (PH) domain of GEP100 and Tyr1139/Tyr1196 of Her2. Tyr1139/Tyr1196 are autonomously phosphorylated, when Her2 is overexpressed. Accordingly, invasive activities mediated by the Her2-GEP100 pathway are not dependent on external factors. Blocking Her2-GEP100 binding, as well as its signaling pathway all inhibit cancer invasive activities. Moreover, our clinical study indicates that co-overexpression of Her2 with GEP100 in primary lung adenocarcinomas of patients is correlated with the presence of their node-metastasis with a statistical significance. Since the GEP100 PH domain interacts with both Her2 and EGFR, targeting this domain may provide novel cancer therapeutics.
  • Ari Hashimoto, Shigeru Hashimoto, Ryo Ando, Kosuke Noda, Eiji Ogawa, Hirokazu Kotani, Mayumi Hirose, Toshi Menju, Masaki Morishige, Toshiaki Manabe, Yoshinobu Toda, Susumu Ishida, Hisataka Sabe
    PLOS ONE 6 (8) e23359  1932-6203 2011/08 [Refereed][Not invited]
     
    Angiogenesis and cancer invasiveness greatly contribute to cancer malignancy. Arf6 and its effector, AMAP1, are frequently overexpressed in breast cancer, and constitute a central pathway to induce the invasion and metastasis. In this pathway, Arf6 is activated by EGFR via GEP100. Arf6 is highly expressed also in human umbilical vein endothelial cells (HUVECs) and is implicated in angiogenesis. Here, we found that HUVECs also highly express AMAP1, and that vascular endothelial growth factor receptor-2 (VEGFR2) recruits GEP100 to activate Arf6. AMAP1 functions by binding to cortactin in cancer invasion and metastasis. We demonstrate that the same GEP100-Arf6-AMAP1-cortactin pathway is essential for angiogenesis activities, including cell migration and tubular formation, as well as for the enhancement of cell permeability and VE-cadherin endocytosis of VEGF-stimulated HUVECs. Components of this pathway are highly expressed in pathologic angiogenesis, and blocking of this pathway effectively inhibits VEGF-or tumor-induced angiogenesis and choroidal neovascularization. The GEP100-Arf6-AMAP1-cortactin pathway, activated by receptor tyrosine kinases, appears to be common in angiogenesis and cancer invasion and metastasis, and provides their new therapeutic targets.
  • Hisataka Sabe
    JOURNAL OF BIOCHEMISTRY 6 149 (6) 633 - 639 0021-924X 2011/06 [Refereed][Not invited]
     
    Contrary to the long believed hypothesis, it is now evident that breast cancer cells can disseminate from the early phases of the oncogenesis; and that such early disseminated cells sometimes survive at the sites of dissemination and may outgrow after a long latency of years and decades. For cancer cells to leave their origin, they must at least transiently loosen their adhesion with adjacent epithelial cells and stroma, and become motile while avoiding anoikis. Such processes resemble epithelial-mesenchymal transdifferentiation (EMT), which normally takes place in situations such as embryogenesis and wound healing. Interestingly, the occurrence of an EMT-like process in breast cancer cells has been implicated in the generation of cancer stem-like cells, in which TGF beta 1 signalling often plays core roles. Here, I discuss the current knowledge regarding cancerous EMT and its signalling pathways with the aim to consider the possible mechanisms of early dissemination, and also the generation of cancer stem-like cells in mammary tumour.
  • Hisataka Sabe, Shigeru Hashimoto, Masaki Morishige, Eiji Ogawa, Ari Hashimoto, Jin-Min Nam, Koichi Miura, Hajime Yano, Yasuhito Onodera
    TRAFFIC 10 (8) 982 - 993 1398-9219 2009/08 [Refereed][Not invited]
     
    Tumors are tissue-specific diseases, and their mechanisms of invasion and metastasis are highly diverse. In breast cancer, biomarkers that specifically correlate with the invasive phenotypes have not been clearly identified. A small GTPase Arf6 primarily regulates recycling of plasma membrane components. We have shown that Arf6 and its effector AMAP1 (DDEF1, DEF1, ASAP1 and centaurin beta 4) are abnormally overexpressed in some breast cancers and used for their invasion and metastasis. Overexpression of these proteins is independent of the transcriptional upregulation of their genes, and occurs only in highly malignant breast cancer cells. We recently identified GEP100 (BRAG2) to be responsible for the Arf6 activation to induce invasion and metastasis, by directly binding to ligand-activated epidermal growth factor receptor (EGFR). A series of our studies revealed that for activation of the invasion pathway of EGFR, it is prerequisite that Arf6 and AMAP1 both are highly overexpressed, and that EGFR is activated by ligands. Pathological analyses indicate that a significant large population of human ductal cancers may utilize the EGFR-GEP100-Arf6-AMAP1 pathway for their malignancy. Microenvironments have been highly implicated in the malignancy of mammary tumors. Our results reveal an aspect of the precise molecular mechanisms of some breast cancers, in which full invasiveness is not acquired just by intracellular alterations of cancer cells, but extracellular factors from microenvironments may also be necessary. Possible translation of our knowledge to cancer therapeutics will also be discussed.
  • Masaki Mori, Hironori Nakagami, Nobutaka Koibuchi, Koichi Miura, Yoichi Takami, Hiroshi Koriyama, Hiroki Hayashi, Hisataka Sabe, Naoki Mochizuki, Ryuichi Morishita, Yasufumi Kaneda
    MOLECULAR BIOLOGY OF THE CELL 20 (13) 3115 - 3124 1059-1524 2009/07 [Refereed][Not invited]
     
    Epithelial-mesenchymal transition (EMT) confers destabilization of cell-cell adhesion and cell motility required for morphogenesis or cancer metastasis. Here we report that zyxin, a focal adhesion-associated LIM protein, is essential for actin reorganization for cell migration in TGF-beta 1-induced EMT in normal murine mammary gland (NMuMG) cells. TGF-beta 1 induced the relocation of zyxin from focal adhesions to actin fibers. In addition, TGF-beta 1 up-regulated zyxin via a transcription factor, Twist1. Depletion of either zyxin or Twist1 abrogated the TGF-beta 1-dependent EMT, including enhanced cell motility and actin reorganization, indicating the TGF-beta 1-Twist1-zyxin signal for EMT. Both zyxin and Twist1 were predominantly expressed in the cardiac atrioventricular canal (AVC) that undergoes EMT during heart development. We further performed ex vivo AVC explant assay and revealed that zyxin was required for the reorganization of actin fibers and migration of the endocardial cells. Thus, zyxin reorganizes actin fibers and enhances cell motility in response to TGF-beta 1, thereby regulating EMT.
  • Zyxin Mediates Actin Fiber Reorganization in Epithelial-Mesenchymal Transition and Contributes to Endocardial Morphogenesis
    Masaki Mori, Hironori Nakagami, Nobutaka Koibuchi, Koichi Miura, Hisataka Sabe, Naoki Mochizuki, Yasufumi Kaneda
    MOLECULAR THERAPY 17 S353 - S353 1525-0016 2009/05 [Refereed][Not invited]
  • Koichi Miura, Jin-Min Nam, Chie Kojima, Naoki Mochizuki, Hisataka Sabe
    MOLECULAR BIOLOGY OF THE CELL 20 (7) 1949 - 1959 1059-1524 2009/04 [Refereed][Not invited]
     
    ADP-ribosylation factor (Arf) 6 activity is crucially involved in the regulation of E-cadherin-based cell-cell adhesions. Erythropoietin-producing hepatocellular carcinoma (Eph)-family receptors recognize ligands, namely, ephrins, anchored to the membrane of apposing cells, and they mediate cell-cell contact-dependent events. Here, we found that Arf6 activity is down-regulated in Madin-Darby canine kidney cells, which is dependent on cell density and calcium ion concentration, and we provide evidence of a novel signaling pathway by which ligand-activated EphA2 suppresses Arf6 activity. This EphA2-mediated suppression of Arf6 activity was linked to the induction of cell compaction and polarization, but it was independent of the down-regulation of extracellular signal-regulated kinase 1/2 kinase activity. We show that G protein-coupled receptor kinase-interacting protein (Git) 1 and noncatalytic region of tyrosine kinase (Nck) 1 are involved in this pathway, in which ligand-activated EphA2, via its phosphorylated Tyr594, binds to the Src homology 2 domain of Nck1, and then via its Src homology 3 domain binds to the synaptic localizing domain of Git1 to suppress Arf6 activity. We propose a positive feedback loop in which E-cadherin-based cell-cell contacts enhance EphA-ephrinA signaling, which in turn down-regulates Arf6 activity to enhance E-cadherin-based cell-cell contacts as well as the apical-basal polarization of epithelial cells.
  • Miyata M, Raven JF, Baltzis D, Koromilas AE, Sabe H
    Cell cycle (Georgetown, Tex.) 20 7 (20) 3273 - 3281 1538-4101 2008/10 [Refereed][Not invited]
  • Mariko Hirano, Shigeru Hashimoto, Shigenobu Yonemura, Hisataka Sabe, Shinichi Aizawa
    JOURNAL OF CELL BIOLOGY 182 (6) 1217 - 1230 0021-9525 2008/09 [Refereed][Not invited]
     
    EPB41L5 belongs to the band 4.1 superfamily. We investigate here the involvement of EPB41L5 in epithelial-mesenchymal transition (EMT) during mouse gastrulation. EPB41L5 expression is induced during TGF beta-stimulated EMT, whereas silencing of EPB41L5 by siRNA inhibits this transition. In EPB41L5 mutants, cell-cell adhesion is enhanced, and EMT is greatly impaired during gastrulation. Moreover, cell attachment, spreading, and mobility are greatly reduced by EPB41L5 deficiency. Gene transcription regulation during EMT occurs normally at the mRNA level; EPB41L5 siRNA does not affect either the decrease in E-cadherin or the increase in integrin expression. However, at the protein level, the decrease in E-cadherin and increase in integrin are inhibited in both EPB41L5 siRNA-treated NMuMG cells and mutant mesoderm. We find that EPB41L5 binds p120ctn through its N-terminal FERM domain, inhibiting p120ctn-E-cadherin binding. EPB41L5 overexpression causes E-cadherin relocalization into Rab5-positive vesicles in epithelial cells. At the same time, EPB41L5 binds to paxillin through its C terminus, enhancing integrin/paxillin association, thereby stimulating focal adhesion formation.
  • Shunsuke Kon, Kenji Tanabe, Toshio Watanabe, Hisataka Sabe, Masanobu Satake
    EXPERIMENTAL CELL RESEARCH 314 (7) 1415 - 1428 0014-4827 2008/04 [Refereed][Not invited]
     
    E-cadherin is a central component of the adherens junction in epithelial cells and continuously undergoes endocytosis via clathrin-coated vesicles and/or caveolae depending on the cell type. In this study, we examined the role of SMAP1, a clathrin-interacting GTPase-activating protein (GAP) for the ADP-ribosylation factor 6 (Arf6) GTPase, in E-cadherin endocytosis. Mardin-Darby canine kidney (MDCK) epithelial cells were used as a model, and SMAP1 localized in the cytoplasm and along the adherens junction where E-cadherin was present. Next, activity of SMAP1 was compared with that of other Arf6GAPs (and/or an effector of Arf6-GTP), namely GIT1 and AMAP2/DDEF2. Overexpression of SMAP1 but not GIT1 nor AMAP2/DDEF2 strongly inhibited basal, as well as phorbolester-induced, internalization of E-cadherin. Notably, AMAP2/DDEF2 rather enhanced the caveolae-mediated incorporation of a membrane protein other than E-cadherin. Thus, in MDCK cells, E-cadherin appeared to be endocytosed solely through SMAP1-regulated clathrin-coated vesicles. Furthermore, MDCK cells overexpressing SMAP1 showed a reduced degree of cell migration compared to untransfected cells, as assessed by wound healing and Transwell assays, and this reduction in migration appeared to be due to the accumulation of E-cadherin at the adherens junction in cells overexpressing SMAP1. Collectively, SMAP1 likely represents a key Arf6GAP in clathrin dependent endocytosis of E-cadherin in MDCK cells. This activity of SMAP1 in E-cadherin turnover may be involved in epithelial organization and/or epithelial-mesenchymal transition. (C) 2007 Elsevier Inc. All rights reserved.
  • Hisataka Sabe, Shigeru Hashimoto, Masaki Morishige, Ari Hashimoto, Eiji Ogawa
    CELL ADHESION & MIGRATION 2 (2) 71 - 73 1933-6918 2008/04 [Refereed][Not invited]
     
    Arf6 and its effector AMAP1 are overexpressed in malignant breast cancer cells, and are involved in their invasion and metastasis. We recently revealed that GEP100, a guanine nucleotide exchanging factor, is responsible for the activation of Arf6 which induces invasion and metastasis. GEP100 associated directly with ligand-activated epidermal growth factor receptor (EGFR) to be activated. Disruption of E-cadherin-mediated cell-cell adhesion is one of the major steps involved in acquisition of invasive phenotypes of most carcinomas. The EGFR-GEP100-Arf6 pathway not only activated matrix invasion activity but also perturbed E-cadherin function. GEP100 was found to be expressed in more than 80% of invasive ductal carcinomas. However, 60% of ductal carcinomas in situ were also positive for GEP100, in which GEP100 was preferentially coexpressed with EGFR in their malignant cases. Microenvionments have been highly implicated in the development of tumor malignancy. Our results reveal an aspect of the precise molecular mechanism of cancer invasion and metastasis, in which full invasiveness is not acquired just by alterations of cancer cells themselves, but their microenvironments may also play pivotal roles.
  • Hajime Yano, Itaru Kobayashi, Yasuhito Onodera, Frederic Luton, Michel Franco, Yuichi Mazaki, Shigeru Hashimoto, Kazuhiro Iwai, Ze'ev Ronai, Hisataka Sabe
    MOLECULAR BIOLOGY OF THE CELL 19 (3) 822 - 832 1059-1524 2008/03 [Refereed][Not invited]
     
    The small GTP-binding protein Arf6 regulates membrane remodeling at cell peripheries and plays crucial roles in higher orders of cellular functions including tumor invasion. Here we show that Fbx8, an F-box protein bearing the Sec7 domain, mediates ubiquitination of Arf6. This ubiquitination did not appear to be linked to immediate proteasomal degradation of Arf6, whereas Fbx8 knockdown caused hyperactivation of Arf6. Expression of Fbx8 protein was substantially lost in several breast tumor cell lines, in which Arf6 activity is pivotal for their invasion. Forced expression of Fbx8 in these cells suppressed their Arf6 activities and invasive activities, in which the F-box and Sec7 domains of Fbx8 are required. Together with the possible mechanism as to how Fbx8-mediated ubiquitination interferes with the functions of Arf6, we propose that Fbx8 provides a novel suppressive control of Arf6 activity through noncanonical ubiquitination. Our results indicate that dysfunction of Fbx8 expression may contribute to the invasiveness of some breast cancer cells.
  • Tobias Bauer, Nami Motosugi, Koichi Miura, Hisataka Sabe, Takashi Hiiragi
    GENESIS 46 (3) 152 - 162 1526-954X 2008/03 [Refereed][Not invited]
     
    Cytokinesis is a complex process that involves dynamic cortical rearrangement. Our recent time-lapse recordings of the mouse egg unexpectedly revealed a high motility of the second polar body (2pb). Experiments to address its underlying mechanism show that neither mechanical compression by the zona pellucida nor the connection via the mid-body is required for the 2pb movement. Time-lapse recordings establish that the 2pb moves together with the cell membrane. These recordings, in which cell surface proteins are labeled with fluorescent latex-microbeads or monovalent antibodies against whole mouse proteins, indicate that the majority of the surface proteins dynamically accumulate in the cleavage furrow at every cell division. Comparable dynamics of the cell surface proteins, and specifically of E-cadherin, are also observed in cultured epithelial cells. The surface protein dynamics are closely correlated with, and dependent on, those of the underlying cortical actin. The cortical actin network may form a scaffold for membrane proteins and thereby transfer them during contractile ring formation toward the cleavage furrow. Immobilization of surface proteins by tetravalent lectin-mediated crosslinking results in the failure of cleavage, demonstrating that the observed protein dynamics are essential for cytokinesis. We propose that dynamic rearrangement of the cell surface proteins is a common feature of cytokinesis, playing a key role in modifying the mechanical properties of the cell membrane during cortical ingression.
  • Masaki Morishige, Shigeru Hashimoto, Eiji Ogawa, Yoshinobu Toda, Hirokazu Kotani, Mayumi Hirose, Shumei Wei, Ari Hashimoto, Atsuko Yamada, Hajime Yano, Yuichi Mazaki, Hiroshi Kodama, Yoshinori Nio, Toshiaki Manabe, Hiromi Wada, Hidenori Kobayashi, Hisataka Sabe
    NATURE CELL BIOLOGY 10 (1) 85 - U70 1465-7392 2008/01 [Refereed][Not invited]
     
    Epidermal growth factor (EGF) receptor ( EGFR) signalling is implicated in tumour invasion and metastasis(1,2). However, whether there are EGFR signalling pathways specifically used for tumour invasion still remains elusive. Overexpression of Arf6 and its effector, AMAP1, correlates with and is crucial for the invasive phenotypes of different breast cancer cells(3-6). Here we identify the mechanism by which Arf6 is activated to induce tumour invasion. We found that GEP100/BRAG2, a guanine nucleotide exchanging factor (GEF) for Arf6, is responsible for the invasive activity of MDA-MB-231 breast cancer cells, whereas the other ArfGEFs are not. GEP100, through its pleckstrin homology domain, bound directly to Tyr1068/1086-phosphorylated EGFR to activate Arf6. Overexpression of GEP100, together with Arf6, caused non-invasive MCF7 cells(7) to become invasive, which was dependent on EGF stimulation. Moreover, GEP100 knockdown blocked tumour metastasis. GEP100 was expressed in 70% of primary breast ductal carcinomas, and was preferentially co-expressed with EGFR in the malignant cases. Our results indicate that GEP100 links EGFR signalling to Arf6 activation to induce invasive activities of some breast cancer cells, and hence may contribute to their metastasis and malignancy.
  • Jin-Min Nam, Yasuhito Onodera, Yuichi Mazaki, Hiroyuki Miyoshi, Shigeru Hashimoto, Hisataka Sabe
    EMBO JOURNAL 26 (3) 647 - 656 0261-4189 2007/02 [Refereed][Not invited]
     
    Expression of AMAP1 correlates well with the invasive phenotypes and malignancy of human primary breast carcinomas. AMAP1 recruits its binding proteins, such as cortactin and paxillin, to sites of Arf6 activation to form invadopodia. A mouse ortholog of AMAP1, ASAP1, is known to bind to CIN85, a binding partner of an E3 ligase, Cbl. Here, we found that CIN85 colocalizes with AMAP1 at invadopodia, and binding of AMAP1 with CIN85 is important for the invasive activities of breast cancer cells, including MDA-MB-231. siRNA-mediated silencing of CIN85, as well as Cbl, also inhibited the invasion. We moreover found that AMAP1 is monoubiquitinated, rather than polyubiquitinated, by virtue of Cbl and provide evidence that the ability of AMAP1 to be monoubiquitinated is important for its involvement in invasion. Our results indicate that CIN85, as well as Cbl, which is a well-known suppressor of growth factor receptor signaling, can be positively involved in tumor invasion, and suggest that a complex epigenetic process is involved in AMAP1 function in breast cancer cell invasion.
  • Hisataka Sabe, Yasuhito Onodera, Yuichi Mazaki, Shigeru Hashimoto
    CURRENT OPINION IN CELL BIOLOGY 18 (5) 558 - 564 0955-0674 2006/10 [Refereed][Not invited]
     
    The identification of several ArfGAP proteins as binding partners of paxillin, an integrin signaling and scaffolding protein, has suggested the existence of molecular links between integrin functions and intracellular traffic, as proposed by MS Bretscher long ago. Among the paxillin-binding ArfGAPs, AMAP1 has recently been strongly implicated in tumor invasion as well as malignancy, owing to its highly augmented expression in tumors and its direct involvement in invasive activities. Another ArfGAP, Git2, was found to be a component of the G beta gamma-mediated directional sensing machinery, while simultaneously playing an essential role in the suppressive control of superoxide production, which is mediated by vesicle transport in GPCR-stimulated neutrophils. These emerging molecular mechanisms may further delineate key processes regulating intracellular traffic as principal controls of cell motility and invasive activities.
  • Expression of AMAP1 provides novel targets to inhibit the invasion of human glioma cells
    Masaki Morishige, Shigeru Hashimoto, Tatsuya Abe, Hidenori Kobayashi, Hisataka Sabe
    NEURO-ONCOLOGY 8 (4) 435 - 435 1522-8517 2006/10 [Refereed][Not invited]
  • Y Mazaki, S Hashimoto, T Tsujimura, M Morishige, A Hashimoto, K Aritake, A Yamada, JM Nam, H Kiyonari, K Nakao, H Sabe
    NATURE IMMUNOLOGY 7 (7) 724 - 731 1529-2908 2006/07 [Refereed][Not invited]
     
    In neutrophils, superoxide anion production generally accompanies chemotaxis and functions in killing invading pathogens. The GIT2 GTPase-activating protein binds to the guanine nucleotide-exchange factor alpha PIX. Here we show that GIT2 was necessary for directional chemotaxis and for the suppression of superoxide production in G protein-coupled receptor-stimulated neutrophils. GIT2 was also necessary for the orientation of superoxide production toward chemoattractant sources. GIT2 suppressed the activity of ADP ribosylation factor 1 and was a component of the G beta gamma subunit-mediated direction-sensing machinery 'downstream' of G protein-coupled receptor signaling. This study establishes a function for GIT2 in linking chemotaxis and superoxide production in neutrophils and shows that loss of GIT2 in vivo leads to an immunodeficient state.
  • S Hashimoto, M Hiroso, A Hashimoto, M Morishige, A Yamada, H Hosaka, KI Akagi, E Ogawa, C Oneyama, T Agatsuma, M Okada, H Kobayashi, H Wada, H Nakano, T Ikegami, A Nakagawa, H Sabe
    PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA 103 (18) 7036 - 7041 0027-8424 2006/05 [Refereed][Not invited]
     
    Invasive potentials of carcinomas greatly contribute to their metastasis, which is a major threat in most cancers. We have recently shown that Arf6 plays a pivotal role in breast cancer invasive activities and identified AMAP1 as an effector of GTP-Arf6 in invasion. Expression of AMAP1 correlates well with invasive phenotypes of primary tumors of the human breast. We also have shown that AMAP1 functions by forming a trimeric protein complex with cortactin and paxillin. In this complex, AMAP1 binds to the src homology 3 (SH3) domain of cortactin via its proline-rich peptide, SKKRPPPPPPGHKRT. SH3 domains are known to bind generally to the proline-rich ligands with a one-to-one stoichiometry. We found that AMAP1/cortactin binding is very atypical in its stoichiometry and interface structure, in which one AMAP1 proline-rich peptide binds to two cortactin SH3 domains simultaneously. We made a cell-permeable peptide derived from the AMAP1 peptide, and we show that this peptide specifically blocks AMAP1/cortactin binding, but not other canonical SH3/proline bindings, and effectively inhibits breast cancer invasion and metastasis. Moreover, this peptide was found to block invasion of other types of cancers, such as glioblastomas and lung carcinomas. We also found that a small-molecule compound, UCS15A, which was previously judged as a weak inhibitor against canonical SH3/proline bindings, effectively inhibits AMAP1/cortactin binding and breast cancer invasion and metastasis. Together with fine structural analysis, we propose that the AMAP1/cortactin complex, which is not detected in normal mammary epithelial cells, is an excellent drug target for cancer therapeutics.
  • Novel regulation of Arf6 activity
    Hajime Yano, Itaru Kobyashi, Shigeru Hashimoto, Hisataka Sabe
    CELL STRUCTURE AND FUNCTION 30 49 - 49 0386-7196 2005/06 [Refereed][Not invited]
  • Regulation of epithelial cell-cell interaction by Arf6 signaling
    Koichi Miura, Hajime Yano, Yasuhito Onodera, Chie Kojima, Shigeru Hashimoto, Hisataka Sabe
    CELL STRUCTURE AND FUNCTION 30 5 - 5 0386-7196 2005/06 [Refereed][Not invited]
  • AJ Woods, T Kantidakis, H Sabe, DR Critchley, JC Norman
    MOLECULAR AND CELLULAR BIOLOGY 25 (9) 3763 - 3773 0270-7306 2005/05 [Refereed][Not invited]
     
    We have previously identified poly(A)-binding protein 1 (PABP1) as a ligand for paxillin and shown that the paxillin-PABP1 complex undergoes nucleocytoplasmic shuttling. By targeting the paxillin-binding subdomain sequences in PABP1, we have generated mutants of PABP1 that do not bind to cellular paxillin. Here we report that paxillin association is necessary for efficient nuclear export of PABP1 and that RNA interference of paxillin drives the nuclear accumulation of PABP1. Furthermore, ablation of paxillin-PABP1 association impeded a number of indices of cell motility including spreading on fibronectin, cell migration on two-dimensional matrices, and transmigration in Boyden chambers. These data indicate that PABP1 must associate with paxillin in order to be efficiently transported from the nucleus to the cytoplasm and that this event is necessary for cells to remodel their focal adhesions during cell migration.
  • Y Onodera, S Hashimoto, A Hashimoto, M Morishige, Y Mazaki, A Yamada, E Ogawa, M Adachi, T Sakurai, T Manabe, H Wada, N Matsuura, H Sabe
    EMBO JOURNAL 24 (5) 963 - 973 0261-4189 2005/03 [Refereed][Not invited]
     
    Identification of the molecular machinery employed in cancer invasion, but not in normal adult cells, will greatly contribute to cancer therapeutics. Here we found that an ArfGAP, AMAP1/PAG2, is expressed at high levels in highly invasive breast cancer cells, but at very low levels in noninvasive breast cancer cells and normal mammary epithelial cells. siRNA-mediated silencing of AMAP1 effectively blocked the invasive activities. AMAP1 expression in human breast primary tumors also indicated its potential correlation with malignancy. Paxillin and cortactin have been shown to colocalize at invadopodia and play a pivotal role in breast cancer invasion. We found that AMAP1 is also localized at invadopodia, and acts to bridge paxillin and cortactin. This AMAP1-mediated trimeric protein complex was detected only in invasive cancer cells, and blocking this complex formation effectively inhibited their invasive activities in vitro and metastasis in mice. Our results indicate that AMAP1 is a component involved in invasive activities of different breast cancers, and provide new information regarding the possible therapeutic targets for prevention of breast cancer invasion and metastasis.
  • S Hashimoto, A Hashimoto, A Yamada, Y Onodera, H Sabe
    GTPASES REGULATING MEMBRANE DYNAMICS 404 216 - 231 0076-6879 2005 [Refereed][Not invited]
     
    The GTPase-activating protein (GAP) domain for Arfs primarily consists of a zinc-finger structure, which is not present in known GAPS for the other Ras-superfamily GTPases. More than 20 genes have been found to encode proteins bearing the ArfGAP domain in the human genome: a number that is much larger than that of the Arf isoforms. Several Arf isoforms, such as Arf1 and Arf6, indeed have been shown to each employ multiple different ArfGAPs for their regulation and function. We have found that two ArfGAPs, namely AMAP1 and AMAP2, exhibit a novel biochemical property of directly and selectively binding to GTP-Arf6 without immediate GAPing activity, while they were previously shown to exhibit efficient catalytic GAPing activities to Arf isoforms except Arf6 in vitro. Such property of AMAPs appears to be important for AMAPs-mediated recruitment of auxiliary molecules, including paxillin, cortactin, amphiphysin, and intersectin, to sites of Arf6 activation. AMAPs thus appear to act as "effectors" rather than simple GAPS in some aspects of Arf6 function. This article presents methods and protocols developed for the functional characterization of AMAPs in Arf6 function. These methods may be applied to other types of ArfGAPs to further clarify the cellular functions of ArfGAPs as well as Arfs.
  • C Kojima, A Hashimoto, Yabuta, I, M Hirose, S Hashimoto, Y Kanaho, H Sumimoto, T Ikegami, H Sabe
    EMBO JOURNAL 23 (22) 4413 - 4422 0261-4189 2004/11 [Refereed][Not invited]
     
    Bin1/M-amphiphysin-II is an amphiphysin-II isoform highly expressed in transverse tubules of adult striated muscle and is implicated in their biogenesis. Bin1 contains a basic unique amino-acid sequence, Exon10, which interacts with certain phosphoinositides such as phosphatidylinositol-4,5- bisphosphate (PI(4,5) P-2), to localize to membranes. Here we found that Exon10 also binds to the src homology 3 (SH3) domain of Bin1 itself, and hence blocks the binding of the SH3 domain to its canonical PxxP ligands, including dynamin. This blockage was released by addition of PI(4,5) P-2 in vitro or in cells overexpressing phosphatidylinositol 4-phosphate 5-kinase. The Exon10-binding interface of the Bin1 SH3 domain largely overlapped with its PxxP-binding interface. We also show that the PLCdelta pleckstrin homology domain, another PI(4,5) P-2-binding module, cannot substitute for Exon10 in Bin1 function in transverse tubule formation, and suggest the importance of the dual biochemical properties of Exon10 in myogenesis. Our results exemplify a novel mechanism of SH3 domain regulation, and suggest that the SH3-mediated protein - protein interactions of Bin1 are regulated by Exon10 so that it may only occur when Bin1 localizes to certain submembrane areas.
  • S Hashimoto, A Hashimoto, A Yamada, C Kojima, H Yamamoto, T Tsutsumi, M Higashi, A Mizoguchi, R Yagi, H Sabe
    JOURNAL OF BIOLOGICAL CHEMISTRY 279 (36) 37677 - 37684 0021-9258 2004/09 [Refereed][Not invited]
     
    Previously we reported that AMAP2/PAG3/Papalpha/ KIAA0400, a GTPase-activating protein ( GAP), acts to antagonize Arf6 function when overexpressed, whereas it was shown to exhibit efficient GAP activities for other Arf isoforms in vitro. Here, we found that AMAP2, through its ArfGAP domain, binds to GTP-Arf6 but not to GDP-Arf6 or other Arfs irrespective of nucleotide status. The majority of AMAP2 was localized to intracellular tubulovesicular structures and redistributed to Arf6-enriched membrane areas upon Arf6 activation. In HeLa cells, Arf6 has been shown to be involved in the clathrin-independent endocytosis of Tac, but not the clathrin-dependent endocytosis of transferrin. We found that Arf6 silencing inhibited the internalization of Tac, but not transferrin, in HeLa cells. Internalization of Tac, but not transferrin, was also significantly inhibited by AMAP2 silencing and overexpression. AMAP2 was moreover found to bind to amphiphysin IIm, a component of the endocytic machinery, via its proline-rich domain. We propose that AMAP2 has dual mechanisms for its function; it exhibits efficient catalytic GAP activity for the class I and II Arfs and yet is involved in the cellular function of the class III Arf without immediate GAP activity. These dual mechanisms of AMAP2 may be important for the cellular function of GTP-Arf6.
  • LY Romanova, S Hashimoto, KO Chay, MV Blagosklonny, H Sabe, JF Mushinski
    JOURNAL OF CELL SCIENCE 117 (17) 3759 - 3768 0021-9533 2004/08 [Refereed][Not invited]
     
    Tyrosine phosphorylation of paxillin regulates actin cytoskeleton-dependent changes in cell morphology and motility in adherent cells. In this report we investigated the involvement of paxillin tyrosine phosphorylation in the regulation of actin cytoskeleton-dependent polarization and motility of a non-adherent IL-3-dependent murine pre-B lymphocytic cell line Baf3. We also assessed the effect of phorbol myristate acetate (PMA), a phorbol ester analogous to those currently in clinical trials for the treatment of leukemia, on paxillin phosphorylation. Using tyrosine-to-phenylalanine phosphorylation mutants of paxillin and phosphospecific antibody we demonstrated that IL-3 stimulated phosphorylation of paxillin tyrosine residues 31 and 118, whereas the tyrosines 40 and 181 were constitutively phosphorylated. Phosphorylation of paxillin residues 31 and 118 was required for cell polarization and motility. In the presence of IL-3, PMA dramatically reduced the phosphorylation of residues 31 and 118, which was accompanied by inhibition of cell polarization and motility. This PMA effect was partially recapitulated by expression of exogenous tyrosine 31 and 118 mutants of paxillin. We also demonstrated that PMA inhibited the IL-3-induced and activation-dependent tyrosine phosphorylation of focal adhesion kinase. Thus, our results indicate that phosphorylation of paxillin tyrosine residues 31 and 118 regulates actin-dependent polarization and motility of pre-B Baf3 cells, both of which could be inhibited by PMA. They also suggest that inhibition of upstream signaling by PMA contributes to the decrease of paxillin phosphorylation and subsequent changes in cell morphology.
  • H Yano, Y Mazaki, K Kurokawa, SK Hanks, M Matsuda, H Sabe
    JOURNAL OF CELL BIOLOGY 166 (2) 283 - 295 0021-9525 2004/07 [Refereed][Not invited]
     
    Integrins can intercommunicate with cadherins. Here, we examined their possible relationship by use of small interfering RNA-mediated protein knockdown in HeLa cells. We found that a subset of integrin signaling molecules, namely Fak and paxillin, but not p130 Crk-associated substrate or proline-rich tyrosine kinase 2, participate in processes regulating N-cadherin-based cell-cell adhesion. Paxillin was found to be required primarily for the recruitment of Fak to robust focal adhesions. Our results suggest that at least some signals involving Fak are linked to a mechanism down-regulating Rac1 activity at the cell periphery, which appears to be important for the formation of N-cadherin-based adhesions in motile cells. Our analyses simultaneously exemplified the essential role of Fak in the maintenance of cell-cell adhesions in collective cell migration, a type of migration occurring in embryonic development and carcinoma invasion.
  • S Hashimoto, Y Onodera, A Hashimoto, M Tanaka, M Hamaguchi, A Yamada, H Sabe
    PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA 101 (17) 6647 - 6652 0027-8424 2004/04 [Refereed][Not invited]
     
    in most human breast cancer cell lines, there is a direct correlation between their in vivo invasive phenotypes and in vitro invasion activities. Here, we found that ADP-ribosylation factor 6 (Arf6) is localized at the invadopodia of the cultured breast cancer cells MDA-MB-231, and its suppression by a small-interfering RNA duplex effectively blocks the invasive activities of the cells, such as invadopodia formation, localized matrix degradation and Matrigel transmigration but not the cell-adhesion activity. We also found that the GTP hydrolysis-defective mutant Arf6(Q67L) and the GTP-binding defective mutant Arf6(T27N) both blocked these invasive activities but not cell adhesion, suggesting the necessity of continued activation and cycling of the Arf6 GTPase cycle in invasion. Among the different human breast cancer cell lines that we examined, cell lines with high invasive activities expressed higher amounts of Arf6 protein than those in weakly invasive and noninvasive cell lines, although no notable correlation was found between Arf6 mRNA expression levels and invasive activities. Moreover, Matrigel-transmigration activity of all of these invasive cells was blocked effectively by an Arf6 small-interfering RNA duplex. Hence, Arf6 appears to be an integral component of breast cancer invasive activities, and we propose that Arf6 and the intracellular machinery regulating Arf6 during invasion should be considered as therapeutic targets for the prevention of breast cancer invasion.
  • H Watanabe, T Shimizu, J Nishihira, R Abe, T Nakayama, M Taniguchi, H Sabe, T Ishibashi, H Shimizu
    JOURNAL OF BIOLOGICAL CHEMISTRY 279 (3) 1676 - 1683 0021-9258 2004/01 [Refereed][Not invited]
     
    Matrix metalloproteinases ( MMPs) are thought to be responsible for dermal photoaging in human skin. In the present study, we evaluated the involvement of macrophage migration inhibitory factor (MIF) in MMP-1 expression under ultraviolet A (UVA) irradiation in cultured human dermal fibroblasts. UVA ( 20 J/cm(2)) up-regulates MIF production, and UVA-induced MMP-1 mRNA production is inhibited by an anti-MIF antibody. MIF ( 100 ng/ml) was shown to induce MMP-1 in cultured human dermal fibroblasts. We found that MIF ( 100 ng/ml) enhanced MMP-1 activity in cultured fibroblasts assessed by zymography. Moreover, we observed that fibroblasts obtained from MIF-deficient mice were much less sensitive to UVA regarding MMP-13 expression than those from wild-type BALB/c mice. Furthermore, after UVA irradiation ( 10 J/cm(2)), dermal fibroblasts of MIF-deficient mice produced significantly decreased levels of MMP-13 compared with fibroblasts of wild-type mice. Next we investigated the signal transduction pathway of MIF. The up-regulation of MMP-1 mRNA by MIF stimulation was found to be inhibited by a PKC inhibitor (GF109203X), a Src-family tyrosine kinase inhibitor ( herbimycin A), a tyrosine kinase inhibitor ( genistein), a PKA inhibitor (H89), a MEK inhibitor (PD98089), and a JNK inhibitor (SP600125). In contrast, the p38 inhibitor (SB203580) was found to have little effect on expression of MMP-1 mRNA. We found that PKC-pan, PKCalpha/betaII, PKCdelta (Thr505), PKCdelta (Ser(643)), Raf, and MAPK were phosphorylated by MIF. Moreover, we demonstrated that phosphorylation of PKCalpha/betaII and MAPK in response to MIF was suppressed by genistein, and herbimycin A as well as by transfection of the plasmid of C-terminal Src kinase. The DNA binding activity of AP-1 was significantly up-regulated 2 h after MIF stimulation. Taken together, these results suggest that MIF is involved in the up-regulation of UVA-induced MMP-1 in dermal fibroblasts through PKC-, PKA-, Src family tyrosine kinase, MAPK-,c-Jun-, and AP-1-dependent pathways.
  • H Sabe
    JOURNAL OF BIOCHEMISTRY 134 (4) 485 - 489 0021-924X 2003/10 [Refereed][Not invited]
     
    Cell movements are essential to life, in a variety of aspects including development, repair and defence processes. Cell migration is a multifactorial process in which a number of distinct events occur simultaneously. Besides its strong appeal towards the basic sciences, the molecular mechanisms of cell migration have long been major targets of oncology, including clinical studies aiming for cancer therapy and prevention. For the further advancement of these studies, as well as for the benefit of its clinical applications, it is important to understand the fundamental machinery and mechanisms regulating cell adhesion and motility. Here the possible roles of a small GTP-binding protein, Arf6, in epithelial cell adhesion and migration, and also in cancer cell invasion, are discussed.
  • C Didier, L Broday, A Bhoumik, S Israeli, S Takahashi, K Nakayama, SM Thomas, CE Turner, S Henderson, H Sabe, Z Ronai
    MOLECULAR AND CELLULAR BIOLOGY 23 (15) 5331 - 5345 0270-7306 2003/08 [Refereed][Not invited]
     
    RNF5 is a RING finger protein found to be important in the growth and development of Caenorhabditis elegans. The search for RNF5-associated proteins via a yeast two-hybrid screen identified a LIM-containing protein in C elegans which shows homology with human paxillin. Here we demonstrate that the human homologue of RNF5 associates with the amino-terminal domain of paxillin, resulting in its ubiquitination. RNF5 requires intact RING and C-terminal domains to mediate paxillin ubiquitination. Whereas RNF5 mediates efficient ubiquitination of paxillin in vivo, protein extracts were required for in vitro ubiquitination, suggesting that additional modifications and/or an associated E3 ligase assist RNF5 targeting of paxillin ubiquitination. Mutant Ubc13 efficiently inhibits RNF5 ubiquitination, suggesting that RNF5 generates polychain ubiquitin of the K63 topology. Expression of RNF5 increases the cytoplasmic distribution of paxillin while decreasing its localization within focal adhesions, where it is primarily seen under normal growth. Concomitantly, RNF5 expression results in inhibition of cell motility. Via targeting of paxillin ubiquitination, which alters its localization, RNF5 emerges as a novel regulator of cell motility.
  • Y Yamamoto, T Maruyama, N Sakai, R Sakurai, A Shimizu, T Hamatani, H Masuda, H Uchida, H Sabe, Y Yoshimura
    MOLECULAR HUMAN REPRODUCTION 8 (12) 1117 - 1124 1360-9947 2002/12 [Refereed][Not invited]
     
    Decidual growth factors and locally produced cytokines are thought to activate specific phosphorylation signalling pathway(s), thereby eliciting a variety of decidual functions. We have previously reported the activation of c-Src tyrosine kinase during ovarian steroid-induced decidualization of cultured human endometrial stromal cells. As chicken c-Src is known to be activated upon dephosphorylation of tyrosine 527 (Y527, corresponding to Y530 in human), we here employed a monoclonal antibody, clone 28, directed against the active form of human c-Src whose Y530 is dephosphorylated, and investigated whether c-Src became dephosphorylated at Y530 and thereby activated during decidualization. We found that the active form of c-Src was up-regulated and demonstrated increased kinase activity during in-vitro decidualization. Immunohistochemistry revealed that decidual cells in early pregnancy decidua were intensely stained with clone 28 when compared with the stromal cells in the non-pregnant endometrium. Moreover, the active form of c-Src translocated from a perinuclear region to the cytoplasm upon decidualization. Thus, the Y530 dephosphorylation, kinase activation, and subcellular translocation of c-Src may be intracellular signalling events associated with decidualization in vivo as well as in vitro.
  • [Ensemble of membrane and cytoskeleton remodelings in cell migration].
    Sabe H
    Seikagaku. The Journal of Japanese Biochemical Society 12 74 1429 - 1440 0037-1017 2002/12 [Refereed][Not invited]
  • A Tsubouchi, J Sakakura, R Yagi, Y Mazaki, E Schaefer, H Yano, H Sabe
    JOURNAL OF CELL BIOLOGY 159 (4) 673 - 683 0021-9525 2002/11 [Refereed][Not invited]
     
    RhoA activity is transiently inhibited at the initial phase of integrin engagement, when Cdc42- and/or Rac1-mediated membrane spreading and ruffling predominantly occur. Paxillin, an integrin-assembly protein, has four major tyrosine phosphorylation sites, and the phosphorylation of Tyr31 and Tyrl 18 correlates with cell adhesion and migration. We found that mutation of Tyr31/118 caused enhanced activation of RhoA and premature formation of stress fibers with substantial loss of efficient membrane spreading and ruffling in adhesion and migration of NMuMG cells. These phenotypes were similar to those induced by RhoA(G14V) in parental cells, and could be abolished by expression of RhoA(T19N), Rac1(G12V), or p190RhoGAP in the mutant-expressing cells. Phosphorylated Tyr31/118 was found to bind to two src homology (SH)2 domains of p120RasGAP, with coprecipitation of endogenous paxillin with p120RasGAP. p190RhoGAP is known to be a major intracellular binding partner for the p120RasGAP SH2 domains. We found that Tyr31/118-phosphorylated paxillin competes with p190RhoGAP for binding to p120RasGAP, and provides evidence that p190RhoGAP freed from p120RasGAP efficiently suppresses RhoA activity during cell adhesion. We conclude that Tyr31/118-phosphorylated paxillin serves as a template for the localized suppression of RhoA activity and is necessary for efficient membrane spreading and ruffling in adhesion and migration of NMuMG cells.
  • T Oshiro, S Koyama, S Sugiyama, A Kondo, Y Onodera, T Asahara, H Sabe, A Kikuchi
    JOURNAL OF BIOLOGICAL CHEMISTRY 277 (41) 38618 - 38626 0021-9258 2002/10 [Refereed][Not invited]
     
    POB1 was previously identified as a RalBP1-binding protein. POB1 and RalBP1 function downstream of small G protein Ral and regulate receptor-mediated endocytosis. To look for additional functions of POB1, we screened for POB1-binding proteins using a yeast two-hybrid method and found that POB1 interacts with mouse ASAP1, which is a human PAG2 homolog. PAG2 is a paxillin-associated protein with ADP-ribosylation factor GTPase-activating protein activity. POB1 formed a complex with PAG2 in intact cells. The carboxyl-terminal region containing the proline-rich motifs of POB1 directly bound to the carboxyl-terminal region including the SH3 domain of PAG2. Substitutions of Pro(423) and Pro(426) with Ala (POB1(PA)) impaired the binding of POB1 to PAG2. Expression of PAG2 inhibited fibronectin-dependent migration and paxillin recruitment to focal contacts of CHO-IR cells. Co-expression with POB1 but not with POB1(PA) suppressed the inhibitory action of PAG2 on cell migration and paxillin localization. These results suggest that POB1 interacts with PAG2 through its proline-rich motif, thereby regulating cell migration.
  • AJ Woods, MS Roberts, J Choudhary, ST Barry, Y Mazaki, H Sabe, SJ Morley, DR Critchley, JC Norman
    JOURNAL OF BIOLOGICAL CHEMISTRY 277 (8) 6428 - 6437 0021-9258 2002/02 [Refereed][Not invited]
     
    Using mass spectrometry we have identified proteins which co-immunoprecipitate with paxillin, an adaptor protein implicated in the integrin-mediated signaling pathways of cell motility. A major component of paxillin immunoprecipitates was poly(A)-binding protein 1, a 70-kDa mRNA-binding protein. Poly(A)-binding protein 1 associated with both the alpha and beta isoforms of paxillin, and this was unaffected by RNase treatment consistent with a protein-protein interaction. The NH2-terminal region of paxillin (residues 54-313) associated directly with poly(A)-binding protein 1 in cell lysates, and with His-poly(A)-binding protein 1 immobilized in microtiter wells. Binding was specific, saturable and of high affinity (K-d of approximate to10 nm). Cell fractionation studies showed that at steady state, the bulk of paxillin and poly(A)-binding protein 1 was present in the "dense" polyribosome-associated endoplasmic reticulum. However, inhibition of nuclear export with leptomycin B caused paxillin and poly(A)-binding protein 1 to accumulate in the nucleus, indicating that they shuttle between the nuclear and cytoplasmic compartments. When cells migrate, poly(A)binding protein 1 colocalized with paxillin-beta at the tips of lamellipodia. Our results suggest a new mechanism whereby a paxillin-poly(A) -binding protein I complex facilitates transport of mRNA from the nucleus to sites of protein synthesis at the endoplasmic reticulum and the leading lamella during cell migration.
  • T Iwasaki, A Nakata, M Mukai, K Shinkai, H Yano, H Sabe, E Schaefer, M Tatsuta, T Tsujimura, N Terada, E Kakishita, H Akedo
    INTERNATIONAL JOURNAL OF CANCER 97 (3) 330 - 335 0020-7136 2002/01 [Refereed][Not invited]
     
    We demonstrated previously that rat ascites hepatoma MMI cells require both lysophosphatidic acid (LPA) and fibronectin (FN) for phagokinetic motility and transcellular migration and that these events are regulated through the RhoA-ROCK pathway. It remains to be elucidated, however, how the signals from both LPA and FN are integrated into cell migration. To examine this, total cellular lysates after stimulation with LPA or FN were subjected to time-course immunoblot analysis with anti-phosphotyrosine antibodies (Abs). Consequently, tyrosine-phosphorylation of paxillin was obviously persistent after stimulation with FN + LPA as compared to after stimulation with either alone. Tyrosine-phosphorylated paxillin comprised 2 components; slowly and fast migrating ones. Immunoblotting of anti-paxillin immunoprecipitates with phosphorylation site-specific Abs revealed the following: tyrosine-phosphorylation was enhanced preferentially on a slowly migrating component after stimulation with FN + LPA; this component contained phosphorylation at both tyrosine residue (Y) 31 and Y118; and phosphorylation of paxillin at Y181 was constitutive and not augmented by stimulation with either FN or LPA. Amiloride, an inhibitor of the Na+/H+ antiporter downstream of ROCK, suppressed cell motility and correspondingly paxillin tyrosine-phosphorylation at both Y31 and Y118. Paxillin phosphorylation weakly induced by FN alone, insufficient for cell migration, was not inhibited by amiloride. These results demonstrate that LPA collaborates with FN for persistent tyrosine phosphorylation of paxillin at both Y31 and Y118, regulated by the Na+/H+ antiporter downstream of ROCK and that this phosphorylated paxillin is essential for MMI cancer cell migration. (C) 2002 Wiley-Liss, Inc.

Books etc

  • 貝淵 弘三, 稲垣 昌樹, 佐邉 壽孝, 松崎 文雄, 貝淵 弘三, 佐邊 壽孝, 稲垣 昌樹, 松崎 文雄 (Joint editor)
    共立出版 2006/11 (ISBN: 4320056426) 812
  • 貝淵 弘三, 稲垣 昌樹, 佐邉 壽孝, 松崎 文雄 
    共立出版 2006
  • Paxillin-associated ArfGAPs: their isoform specificities and roles in coordination.
    'Protein and Cell Regulation' vol.1. 'ARF family GTPases(ed. R. Kahn; Series editors: A. Ridney, J. Frampton; Kluwer Academic Publishers). 2003
  • 細胞-細胞,細胞-基質の相互作用
    岩波講座,現代医学の基礎2 "分子・細胞の生物学(]G0002[)"岩波書店 2000
  • 細胞接着を制御するシグナル
    医学&サイエンスミリーズ 羊土社 1999
  • 細胞接着形成と細胞内小胞輸送活性
    細胞工学 1999
  • TLCによる蛋白質の二次元マッピング
    タンパク質実験法(羊土社) 1996
  • 佐邉 壽孝, 月田 承一郎 
    羊土社 1996 (ISBN: 4897060524)
  • 注目されるシグナル伝達分子:バキシリン(共著)
    実験医学(羊土社) 1995
  • Crk, paxillin
    "用語でまなぶ最新バイオサイエンスライブラリ-"(山本雅編、羊土社) 1995
  • 遺伝子導入法 -リンパ球細胞を中心にして- "DEAE-dextran法"
    免疫学実験操作法(南江堂) 1995
  • Analysis of Csk SH2 binding to tyrosine phosphorylated proteins in c-Src suppression and mitotic activation.(共著)
    Proc.Natl.Acad.Sci.USA 1994
  • シグナルとストレスの起源(共著)
    レドックス制御 最新医学(最新医学社) 1994
  • Analysis of Csk SH2 binding to tyrosine phosphorylated proteins in c-Src suppression and mitotic activation.(共著)
    Proc.Natl.Acad.Sci.USA 1994
  • Activation of c-Src in cells bearing v-Crk and its suppression by Csk.(共著)
    Mol.Cell.Biol. 1992
  • v-Crk発現細胞におけるc-Srcの活性化とCSKによるその抑制
    癌抑制遺伝子(野田 亮編、羊土社)実験医学増刊号 1992
  • Activation of c-Src in cells bearing v-Crk and its suppression by Csk.(共著)
    Mol.Cell.Biol. 1992
  • 受容体の性質と測定 -結合測定法-
    新生化学実験講座(日本生化学会編)(東京化学同人)(共著) 1991
  • インタ-ロイキン受容体遺伝子の単離法(共著)
    新生化学実験講座(日本生化学会編)(東京化学同人)(共著) 1989
  • インタ-ロイキン2とそのレセプタ-
    シリ-ズ分子生物学の進歩(日本分子生物学会編、丸善) 1989
  • Interleukin 2 receptor.(共著)
    The molecular Biology of Receptor (eds.A.D.Strosberg, Ellis Horwood Ltd., Chichester, England) 1988
  • CTLL-2 バイオテクノロジ-素材としての培養細胞-遺伝移入と発現-(瀬野旱二編)
    蛋白質・核酸・酵素(共立出版) 1988
  • Interleukin 2 receptor.(共著)
    The molecular Biology of Receptor (eds.A.D.Strosberg, Ellis Horwood Ltd., Chichester, England) 1988
  • Growth factors and receptors of lymphocytes.(共著)
    Acta Neurochiurgica (eds.K.Sano and S.Ishii, Springer-Verlag, NY, USA) 1987
  • Growth factors and receptors of lymphocytes.(共著)
    Acta Neurochiurgica (eds.K.Sano and S.Ishii, Springer-Verlag, NY, USA) 1987
  • Interleukin 2 receptor ; structure, function, and expression.(共著)
    Cold Spring Harbor Symposia on Quantitative Biology 1986
  • Interleukin 2 receptor ; structure, function, and expression.(共著)
    Cold Spring Harbor Symposia on Quantitative Biology 1986
  • Properties of human IL2 receptor expressed on non-lymphoid cells by cDNA transfection.(共著)
    Mol.Biol.Med. 1984
  • Properties of human IL2 receptor expressed on non-lymphoid cells by cDNA transfection.(共著)
    Mol.Biol.Med. 1984

Conference Activities & Talks

  • SABE Hisataka
    第76回日本癌学会学術総会  2017/09  パシフィコ横浜
  • The Arf6 pathway: a central pathway driving mesenchymal malignancies and drug-resistance of refractory cancers  [Invited]
    SABE Hisataka
    愛知県がんセンター研究所 特別招聘セミナー  2017/07  愛知県がんセンター研究所
  • Hisataka Sabe, Shigeru Hashimoto, Ari Hashimoto, Yutaro Otsuka, Haruka Handa, Tsukasa Oikawa, Yasuhito Onodera
    第68回日本細胞生物学会大会  2016/06  京都テルサ
  • Hisataka Sabe, Ari Hashimoto, Shigeru Hashimoto, Hirokazu Sugino, Ayumu Yoshikawa, Haruka Handa, Yutaro Otsuka, Tsukasa Oikawa, Shuji Mikami, Mototsugu Oya
    The 4th Global Cancer Genomics Consortium Symposium  2014/11  芝蘭会館
  • 佐邊 壽孝
    社団法人がんと炎症・代謝研究会理事会  2014/05
  • SABE Hisataka
    第72回日本癌学会学術総会  2013/10
  • 癌の再発予測と再発部治療  [Invited]
    佐邊 壽孝
    第8回神戸がん研究会  2013/07
  • RTKs-GEP100-Arf6-AMAP1 pathway mediates cancerous EMT in response to mutant p53 and TGFβ1 signaling  [Invited]
    佐邊 壽孝
    東北大学グローバルCOEシンポジウム  2013/02
  • P53変異におるリガンド依存性癌浸潤転移シグナル経路の創出  [Invited]
    佐邊 壽孝
    第4回kansai Cardiovascular & Metabolic Frontier  2012/10
  • Mutant-p53 generates an Arf6 pathway to be activated by TGFb1 and RTKs to promote invasion  [Invited]
    佐邊 壽孝
    第22回北九州がんセミナー  2012/10
  • RTKs-GEP100-Arf6-AMAP1 pathway mediates cancerous EMT in response to mutant p53 and TGFβ1 signaling  [Invited]
    佐邊 壽孝
    第71回日本癌学会学術総会  2012/09
  • 乳癌における発癌初期段階からの前身播種の分子機構とその予測バイオマーカー  [Invited]
    佐邊 壽孝
    平成24年度JMTO臨床試験ワークショップ  2012/06
  • RTKs-GEP100-Arf6-AMAP1 pathway mediates cancerous EMT in response to mutant p53 and TGFβ1 signaling  [Invited]
    佐邊 壽孝
    第8回OOTR年次学会  2012/04
  • p53 mutation and TGFβ signaling culminate in cancer invasiveness via GEP100-Arf6-AMAP1 pathway  [Invited]
    佐邊 壽孝
    平成23年度ライフサイエンスイノベージョン推進機構セミナー 第371回 学内セミナー  2012/02
  • p53 mutation and TGFβ signaling culminate in cancer invasiveness via GEP100-Arf6-AMAP1 pathway.  [Not invited]
    佐邊 壽孝
    Cell Migration in Biology and Medicine  2012/01
  • p53 mutation and TGFβ signaling culminate in cancer invasiveness via GEP100-Arf6-AMAP1 pathway.  [Not invited]
    佐邊 壽孝
    第34回日本分子生物学会年会  2011/12
  • 癌の初期播種と転移部における癌の休眠と再発:癌的EMTから考える  [Not invited]
    佐邊 壽孝
    JSTさきがけ研究同窓会発表会  2010/10
  • The Arf6-AMAP1 Pathway Contributes to Cancerous EMT  [Not invited]
    佐邊 壽孝
    第69回日本癌学会学術総会  2010/09
  • The Arf6-AMAP1 Pathway Contributes to Cancerous EMT.  [Not invited]
    佐邊 壽孝
    第30回札幌国際がんシンポジウム  2010/06
  • 転移部における癌の休眠と再発:癌的EMTから考える  [Not invited]
    佐邊 壽孝
    第62回日本細胞生物学会大会  2010/05
  • Integrin activation and E-cadherin inactivation by the Arf6 pathway specific to breast cancer invasion and metastasis.  [Not invited]
    佐邊 壽孝
    第6回OOTR年次学会  2010/02
  • メンブレントラフィックと癌浸潤形質獲得機序  [Invited]
    佐邊 壽孝
    第82回日本生化学会  2009/10
  • Integrin activation and E-cadherin inactivation by the Arf6 pathway specific to breast cancer invasion and metastasis.  [Invited]
    SABE Hisataka
    2009 FEBS workshop  2009/09
  • 癌の浸潤形質獲得機序:その組織特異性についての考察  [Invited]
    佐邊 壽孝
    生化学会支部例会  2009/07

MISC

  • 真崎雄一, 小野寺康仁, 東恒仁, 堀之内孝広, 及川司, 佐邊壽孝  日本細胞生物学会大会(Web)  69th-  ROMBUNNO.T8‐11(P1‐077) (WEB ONLY)  2017  [Not refereed][Not invited]
  • p53はEZH2と機能的に競合することで上皮性維持に寄与する
    及川 司, 大塚 勇太郎, 小野寺 康仁, 半田 悠, 橋本 あり, 橋本 茂, 鈴木 穣, 佐邊 壽孝  日本癌学会総会記事  75回-  J  -3030  2016/10  [Not refereed][Not invited]
  • Y. Komiya, Y. Onodera, M. Kuroiwa, S. Nomimura, Y. Kubo, J-M Nam, K. Kajiwara, S. Nada, C. Oneyama, H. Sabe, M. Okada  ONCOGENESIS  5-  2016/09  [Not refereed][Not invited]
     
    Epithelial tumor cells often acquire malignant properties, such as invasion/metastasis and uncontrolled cell growth, by undergoing epithelial-mesenchymal transition (EMT). However, the mechanisms by which EMT contributes to malignant progression remain elusive. Here we show that the Rho guanine nucleotide exchange factor (GEF) ARHGEF5 promotes tumor malignancy in a manner dependent on EMT status. We previously identified ARHGEF5, a member of the Dbl family of GEFs, as a multifunctional mediator of Src-induced cell invasion and tumor growth. In the present study, ARHGEF5 was upregulated during tumor growth factor-beta-induced EMT in human epithelial MCF10A cells, and promoted cell migration by activating the Rho-ROCK pathway. ARHGEF5 was necessary for the invasive and in vivo metastatic activity of human colorectal cancer HCT116 cells. These findings underscore the crucial role of ARHGEF5 in cell migration and invasion/metastasis. An in vivo tumorigenesis assay revealed that ARHGEF5 had the potential to promote tumor growth via the phosphatidylinositol 3-kinase (PI3K) pathway. However, ARHGEF5 was not required for tumor growth in epithelial-like human colorectal cancer HCT116 and HT29 cells, whereas the growth of mesenchymal-like SW480 and SW620 cells depended on ARHGEF5. Induction of EMT by tumor necrosis factor-a or Slug in HCT116 cells resulted in the dependence of tumor growth on ARHGEF5. In these mesenchymal-like cells, Akt was activated via ARHGEF5 and its activity was required for tumor growth. Analysis of a transcriptome data set revealed that the combination of ARHGEF5 upregulation and E-cadherin downregulation or Snail upregulation was significantly correlated with poor prognosis in patients with colorectal cancers. Taken together, our findings suggest that EMT-induced ARHGEF5 activation contributes to the progression of tumor malignancy. ARHGEF5 may serve as a potential therapeutic target in a subset of malignant tumors that have undergone EMT.
  • メバロン酸経路阻害剤スタチンはArf6経路を高発現する癌に有効である
    橋本 あり, 橋本 茂, 及川 司, 大塚 勇太郎, 半田 悠, 小野寺 康仁, 佐邊 壽孝  日本生化学会大会プログラム・講演要旨集  89回-  [1P  -268]  2016/09  [Not refereed][Not invited]
  • 小野寺康仁, 堀川芽衣, 佐邊壽孝  がんと代謝研究会プログラム&抄録集  4th-  44  2016/07  [Not refereed][Not invited]
  • 代謝と細胞動態 細胞動態・運命決定を司る内なるチカラ 代謝と輸送および区画化を介したがん形質の誘導
    小野寺 康仁, Bissell Mina, 佐邊 壽孝  日本細胞生物学会大会講演要旨集  68回-  21  -21  2016/05  [Not refereed][Not invited]
  • 古川聖太郎, 橋本あり, 橋本茂, 小野寺康仁, 及川司, 大塚勇太郎, 佐邊壽孝, 平野聡  日本外科学会定期学術集会(Web)  116th-  PS-002-2 (WEB ONLY)  -002  2016/04  [Not refereed][Not invited]
  • 小野寺康仁, BISSELL Mina, 佐邊壽孝  日本細胞生物学会大会(Web)  68th-  ROMBUNNO.S11‐2 (WEB ONLY)  2016  [Not refereed][Not invited]
  • 橋本あり, 橋本茂, 及川司, 大塚勇太郎, 半田悠, 小野寺康仁, 佐邊壽孝  日本生化学会大会(Web)  89th-  ROMBUNNO.1P‐268 (WEB ONLY)  2016  [Not refereed][Not invited]
  • 小野寺康仁, 南ジンミン, 白土博樹, 佐邊壽孝  日本放射線影響学会大会抄録(Web)  59th-  ROMBUNNO.W12‐1 (WEB ONLY)  2016  [Not refereed][Not invited]
  • p53はエピジェネティック制御を介して上皮性を維持する
    及川 司, 小野寺 康仁, 大塚 勇太郎, 半田 悠, 橋本 あり, 橋本 茂, 鈴木 穣, 佐邊 壽孝  日本生化学会大会・日本分子生物学会年会合同大会講演要旨集  88回・38回-  [2P1108]  -[2P1108]  2015/12  [Not refereed][Not invited]
  • Arf6-AMAP1経路によるROS制御は乳癌の放射線抵抗性に寄与する
    小野寺 康仁, 南 ジンミン, 及川 司, 白土 博樹, 佐邊 壽孝  日本癌学会総会記事  74回-  E  -1049  2015/10  [Not refereed][Not invited]
  • 放射線照射後の乳腺上皮細胞の3次元構造維持に関わる分子機序の解析
    南 ジンミン, 小野寺 康仁, 佐邊 壽孝, 白土 博樹  日本癌学会総会記事  74回-  P  -2343  2015/10  [Not refereed][Not invited]
  • Grb2はEGF刺激によるGEP100-Arf6経路活性化を介した腫瘍浸潤転移能を促進する
    毛受 暁史, 今村 直人, 祢里 真也, 長 博之, 中西 崇雄, 志熊 啓, 曽和 晃正, 園部 誠, 佐邊 壽孝, 伊達 洋至  日本癌学会総会記事  74回-  P  -2121  2015/10  [Not refereed][Not invited]
  • 腎盂尿管癌におけるEPB4.1L5発現の意義
    大門 達明, 小坂 威雄, 菊地 栄次, 三上 修治, 宮崎 保匡, 橋本 あり, 橋本 茂, 水野 隆一, 宮嶋 哲, 岡田 保典, 佐邊 壽孝, 大家 基嗣  日本癌学会総会記事  74回-  P  -3251  2015/10  [Not refereed][Not invited]
  • Arf6-AMAP1経路によるROS制御は乳癌の放射線抵抗性に寄与する
    小野寺 康仁, 南 ジンミン, 及川 司, 白土 博樹, 佐邊 壽孝  日本癌学会総会記事  74回-  E  -1049  2015/10  [Not refereed][Not invited]
  • 放射線照射後の乳腺上皮細胞の3次元構造維持に関わる分子機序の解析
    南 ジンミン, 小野寺 康仁, 佐邊 壽孝, 白土 博樹  日本癌学会総会記事  74回-  P  -2343  2015/10  [Not refereed][Not invited]
  • Grb2はEGF刺激によるGEP100-Arf6経路活性化を介した腫瘍浸潤転移能を促進する
    毛受 暁史, 今村 直人, 祢里 真也, 長 博之, 中西 崇雄, 志熊 啓, 曽和 晃正, 園部 誠, 佐邊 壽孝, 伊達 洋至  日本癌学会総会記事  74回-  P  -2121  2015/10  [Not refereed][Not invited]
  • 腎盂尿管癌におけるEPB4.1L5発現の意義
    大門 達明, 小坂 威雄, 菊地 栄次, 三上 修治, 宮崎 保匡, 橋本 あり, 橋本 茂, 水野 隆一, 宮嶋 哲, 岡田 保典, 佐邊 壽孝, 大家 基嗣  日本癌学会総会記事  74回-  P  -3251  2015/10  [Not refereed][Not invited]
  • 小野寺康仁, 南ジンミン, 白土博樹, BISSELL Mina, 佐邊壽孝  がんと代謝研究会プログラム&抄録集  3rd-  44  2015  [Not refereed][Not invited]
  • Arf6-AMAP1経路による酸化還元状態の恒常性維持は乳癌の放射線抵抗性に寄与する(Robust redox homeostasis mediated by Arf6-AMAP1 pathway confers resistance to ionizing radiation in breast cancer)
    小野寺 康仁, 南 ジンミン, 及川 司, 白土 博樹, 佐邊 壽孝  日本癌学会総会記事  73回-  E  -3010  2014/09  [Not refereed][Not invited]
  • EZH2発現亢進により創出されるArf6を中心とした間葉浸潤に特化した分子装置は腎癌の予後不良に関与する(EZH2 generates Arf6-based mesenchymal invasion machinery that is central to poor prognosis of renal cancer)
    橋本 茂, 杉野 弘和, 橋本 あり, 吉河 歩, 及川 司, 半田 悠, 大家 基嗣, 三上 修治, 佐邊 壽孝  日本癌学会総会記事  73回-  J  -2075  2014/09  [Not refereed][Not invited]
  • 乳癌において変異p53がリガンド反応性の間葉型浸潤分子装置を創出する機序(TP53 alterations generate Arf6-based mesenchymal invasion pathway that is activated by RTKs and TGFβ1 in breast cancer)
    橋本 あり, 橋本 茂, 杉野 弘和, 吉河 歩, 及川 司, 小野寺 康仁, 半田 悠, 大塚 勇太郎, 岩見 昴亮, 小根山 千歳, 岡田 雅人, 福田 光則, 佐邊 壽孝  日本癌学会総会記事  73回-  J  -2077  2014/09  [Not refereed][Not invited]
  • p53は間葉系形質を持つ乳がん細胞に上皮系形質を再獲得させる(p53 recalls epithelial memory in mammary cancer cells with mesenchymal phenotypes)
    及川 司, 小野寺 康仁, 橋本 あり, 橋本 茂, 佐邊 壽孝  日本癌学会総会記事  73回-  P  -1159  2014/09  [Not refereed][Not invited]
  • 乳癌における放射線照射後の浸潤能獲得過程に関わる分子機序の解析(Analysis of molecular mechanism involved in invasiveness of radiation treated breast cancer cells)
    南 ジンミン, 小野寺 康仁, 佐邊 壽孝, 白土 博樹  日本癌学会総会記事  73回-  P  -1450  2014/09  [Not refereed][Not invited]
  • Shigeru Hashimoto, Ari Hashimoto, Hirokazu Sugino, Ayumu Yoshikawa, Haruka Handa, Masanao Yoshino, Yutaro Otsuka, Hisataka Sabe  Ras Superfamily Small G Proteins: Biology and Mechanisms 2: Transport  253  -274  2014/05/01  [Not refereed][Not invited]
     
    © 2014 Springer International Publishing Switzerland. All rights reserved.While Arf-family small GTPases (Arf-GTPases) consist of 5 members in humans, 31 human genes have been identified that encode proteins bearing the GTPase-activating protein (GAP) domain for Arf-GTPases. Interestingly, Arf1, the first identified Arf, was shown to substantially lack intrinsic GTPase activity, which other Ras-superfamily members of small GTPases generally bear. Likewise, ArfGAP domains primarily consist of zinc-finger structures, and do not resemble GAP domains for other small GTPases. Arfs primarily function in intracellular vesicle/membrane trafficking. A general model shows that Arfs play roles in membrane budding, in which GTP-Arfs recruit coatomer proteins to generate and maintain membrane curvature to initiate the budding. Coatomers are thought to be separated from Arf-mediated vesicles before they reach the target membrane, while this separation may or may not be coupled with the GTP hydrolysis activity. We have shown that several ArfGAPs, such as AMAP1 and AMAP2, have the ability to bind stably to GTP-Arf6, without immediate GTP hydrolysis. They each contain a BAR domain and hence may act as coatomers for Arf-mediated vesicles. These ArfGAPs moreover act to recruit their binding proteins to sites of Arf6 activation, which are not coatomer components. These findings have amended the classical, general model of the functions of ArfGAPs, as well as Arf-GTPases. In this review, we will describe the recent information revealed about ArfGAPs, with the aim to decipher and discuss their fundamental roles.
  • 新規NFκB抑制メカニズムとしてのAMAP1
    芦田 昇, Nguyen Tien Dat, 岸畑 雅子, 吉川 歩, 佐邊 壽孝, 亀井 加恵子, 荒井 秀典, 北 徹, 横出 正之, 木村 剛  日本臨床分子医学会学術総会プログラム・抄録集  51回-  100  -100  2014/04  [Not refereed][Not invited]
  • K. Miura, Y. Wakayama, M. Tanino, Y. Orba, H. Sawa, M. Hatakeyama, S. Tanaka, H. Sabe, N. Mochizuki  Oncogene  32-  5292  -5301  2013/11/07  [Not refereed][Not invited]
     
    Shp2 is a positive regulator for Erk activation downstream of receptor tyrosine kinases for growth factors. It has been controversial how Shp2 induces Erk activation. We here demonstrate that EphA2 is responsible for Shp2-mediated Erk activation by phosphorylating Tyr542 and Tyr580 of Shp2 in the cells stimulated with growth factors. In NMuMG mammary epithelial cells stimulated with hepatocyte growth factor (HGF), HGF-dependent Erk phosphorylation was prolonged only in the presence of EphA2. This Erk activation paralleled the phosphorylation of Tyr542/580 of Shp2 and the association of Grb2 with Shp2, suggesting the positive signal involving Grb2 signal to activate Ras-Erk pathway. Immunohistochemical studies of mammary cancer specimens revealed that the cancer progression was associated with both Tyr580 phosphorylation of Shp2 and increased expression of EphA2, which were also correlated with increased Erk phosphorylation. Overexpression of either Shp2Thr468Met (a phosphatase- defective mutant found in Lentigines, Electrocardiographic abnormalities, Ocular hypertelorism, Pulmonary stenosis, Abnormal genitalia, Retardation of growth and sensorineural Deafness (LEOPARD) syndrome) or Shp2Asn308Asp (a phosphatase-active mutant found in Noonan syndrome) with EphA2 exhibited comparable activation of Erk and stronger activation than wild-type Shp2, suggesting the phosphatase-independent Erk activation. Expression of Shp2Thr468Met with Tyr542/580Phe mutations resulted in the suppression of Erk activation. Phosphatase-active and -inactive, and wild-type Shp2s bound equally to Grb2, suggesting that phosphorylation of Tyr542/580 of Shp2 was essential but not sufficient for Shp2-mediated Erk activation. We found that Gab1 (Grb2-associated binder 1) was involved in the mutant Shp2-mediated Erk activation. Zebrafish injected with Shp2Thr468Met mRNA showed cardiac edema, whereas those depleted of EphA2b showed less phenotype, suggesting that EphA2 might partly account for the phenotype of LEOPARD syndrome. Collectively, tyrosine phosphorylation of Shp2 by EphA2 contributes to the phosphatase-independent Shp2-mediated activation of Erk and might be involved in Shp2-associated diseases. © 2013 Macmillan Publishers Limited.
  • 癌放射線治療への分子生物学的アプローチ 変異p53が放射線抵抗性に根幹的な間葉型浸潤経路を創出する機構(Toward the improvement of radiotherapy: Approaches from the molecular biological point of view Mechanisms by which oncogenic mutant-p53 generates mesenchymal invasive pathway pivotal to a radiation resis
    佐邊 壽孝, 橋本 あり, 橋本 茂, 小野寺 康仁, 及川 司, Nam Jin-Min, 小根山 千歳, 杉野 弘和, 吉河 歩, 大塚 勇太郎, 半田 悠, 芳野 正修, 岡田 雅人  日本癌学会総会記事  72回-  64  -64  2013/10  [Not refereed][Not invited]
  • 放射線照射後の乳癌再発に関わるシグナルの解析(Possible mechanisms of non-invasive to invasive phenotypic conversion of breast cancer cells upon radiation)
    南 ジンミン, 小野寺 康仁, 石川 正純, 佐邊 壽孝, 白土 博樹  日本癌学会総会記事  72回-  219  -219  2013/10  [Not refereed][Not invited]
  • 細胞が持つリサイクルシステム研究の新展開 p53変異によるGEP100-Arf6-AMAP1経路の活性化と乳癌の浸潤形質獲得
    橋本 茂, 橋本 あり, 小根山 千歳, 吉河 歩, 杉野 弘和, 半田 悠, 芳野 正修, 大塚 勇太郎, 小野寺 康仁, 岡田 雅人, 佐邊 壽孝  日本生化学会大会プログラム・講演要旨集  86回-  2S04a  -3  2013/09  [Not refereed][Not invited]
  • 橋本茂, 橋本あり, 小根山千歳, 吉河歩, 杉野弘和, 半田悠, 芳野正修, 大塚勇太郎, 小野寺康仁, 岡田雅人, 佐邊壽孝  日本生化学会大会(Web)  86th-  2S04A-3 (WEB ONLY)  2013  [Not refereed][Not invited]
  • EZH2によるmiR-203発現抑制が乳癌浸潤に中枢的であるArf6-AMAP1経路創出に関わる(EZH2-mediated downregulation of miR-203 generates the Arf6-AMAP1 pathway pivotal for breast cancer invasiveness)
    杉野 弘和, 橋本 茂, 橋本 あり, 吉河 歩, 半田 悠, 佐邊 壽孝  日本癌学会総会記事  71回-  87  -87  2012/08  [Not refereed][Not invited]
  • がんの浸潤・転移に関与するGEP100-Arf6-AMAP1経路とc-Metシグナルとの相互作用(GAB1 links c-Met signaling with GEP100-Arf6-AMAP1 pathway to promote breast cancer invasiveness)
    吉河 歩, 橋本 茂, 橋本 あり, 杉野 弘和, 大塚 勇太郎, 味藤 静, 半田 悠, 佐邊 壽孝  日本癌学会総会記事  71回-  87  -87  2012/08  [Not refereed][Not invited]
  • 乳がん Arf6経路が変異p53とTGFbeta1シグナルに応答して癌的EMTを引き起こす(Topics of breast cancer RTKs-GEP100-Arf6-AMAP1 pathway mediates cancerous EMT in response to mutant p53 and TGFbeta1 signaling)
    佐邊 壽孝  日本癌学会総会記事  71回-  267  -267  2012/08  [Not refereed][Not invited]
  • 非浸潤性乳管癌の3次元培養細胞モデルにおける放射線の影響と再発に関わる分子機序の解析(Targeting integrin suppresses invasive recurrence in a 3D model of radiation treated ductal carcinoma in situ)
    南 ジンミン, Kazi Ahmed, Sylvain Costes, Hui Zhang, 佐邊 壽孝, 白土 博樹, Catherine Park  日本癌学会総会記事  71回-  375  -375  2012/08  [Not refereed][Not invited]
  • 乳癌浸潤に中枢的なArf6経路は変異p53により創出される(Mutant-p53 generates GEP100-Arf6-AMAP1 pathway to promote breast cancer cell invasiveness in response to TGFbeta1)
    橋本 あり, 橋本 茂, 吉河 歩, 杉野 弘和, 半田 悠, 味藤 静, 佐藤 宏紀, 大塚 勇太郎, 芳野 日南子, 南 ジンミン, 小野寺 康仁, 佐邊 壽孝  日本癌学会総会記事  71回-  399  -399  2012/08  [Not refereed][Not invited]
  • 癌浸潤におけるAMAP1-PRKD2複合体によるインテグリンリサイクリングとその制御機構(beta1 integrin recycling via AMAP1-PRKD2 complex regulated by small GTPases in cancer invasion)
    小野寺 康仁, 南 ジンミン, 橋本 あり, 白土 博樹, 橋本 茂, 佐邊 壽孝  日本癌学会総会記事  71回-  421  -421  2012/08  [Not refereed][Not invited]
  • 橋本あり, 橋本茂, 吉河歩, 杉野弘和, 半田悠, 木下留美子, 畑中佳奈子, 三上修治, 谷野美智枝, 味藤静, 佐藤宏紀, 大塚勇太郎, 芳野日南子, 加戸由加里, NAM Jin‐Min, 小野寺康仁, 田中伸哉, 白土博樹, 佐邊壽孝  日本分子生物学会年会プログラム・要旨集(Web)  35th-  2W10II-1 (WEB ONLY)  2012  [Not refereed][Not invited]
  • 橋本茂, 杉野弘和, 橋本あり, 吉河歩, 大塚勇太郎, 芳野正修, 半田悠, 佐邊壽孝  日本分子生物学会年会プログラム・要旨集(Web)  35th-  2P-0166 (WEB ONLY)  2012  [Not refereed][Not invited]
  • EGF刺激による乳癌細胞浸潤におけるAMAP1の詳細な作用機構(AMAP1 promotes β1 integrin recycling via PRKD2 and Rab5c in EGF-induced invasion of breast cancer cells)
    小野寺 康仁, 南 ジンミン, 橋本 茂, 橋本 あり, 白土 博樹, 佐邊 壽孝  日本癌学会総会記事  70回-  37  -38  2011/09  [Not refereed][Not invited]
  • 変異p53はArf6活性化経路を介した浸潤獲得形質に必須である(Mutant p53 is essential for TGFβ1-induced breast cancer cell invasiveness via activation of GEP100-Arf6-AMAP1 pathway)
    橋本 あり, 橋本 茂, 大塚 勇太郎, 吉河 歩, 杉野 弘和, 半田 悠, 南 ジンミン, 佐藤 宏紀, 福田 諭, 小野寺 康仁, 佐邊 壽孝  日本癌学会総会記事  70回-  38  -38  2011/09  [Not refereed][Not invited]
  • TGFβ及び低酸素によるArf6活性化を介した癌浸潤形質獲得におけるエピジェネティック因子の関与(EZH2 is essential to Arf6 activation necessary for TGFβ1- and hypoxia-induced invasiveness of breast cancer cells)
    橋本 茂, 橋本 あり, 小野寺 康仁, 大塚 勇太郎, 吉河 歩, 杉野 弘和, 半田 悠, 佐藤 宏紀, 福田 諭, 毛受 暁史, 佐邊 壽孝  日本癌学会総会記事  70回-  77  -77  2011/09  [Not refereed][Not invited]
  • 細胞骨格・細胞運動・細胞移動 GBF1のSec7ドメインのHomology Downstreamは、化学走化性とスーパーオキシド産生に重要な役割を果たすArf活性を持つGPCRシグナル伝達とリンクするホスファチジルイノシトールリン酸と結合する(Cytoskeleton/Cell motility/Cell migration The Homology Downstream of Sec7 domain of GBF1 binds to phosphatidyl inositol phosphates to
    真崎 雄一, 佐邊 壽孝  日本細胞生物学会大会講演要旨集  63回-  116  -116  2011/05  [Not refereed][Not invited]
  • 癌の悪性化における糖代謝と小胞輸送の役割(Glucose metabolism and intracellular trafficking in tumor malignancy)
    小野寺 康仁, 南 ジンミン, 橋本 茂, 橋本 あり, 佐邊 壽孝, Bissell Mina  日本細胞生物学会大会講演要旨集  63回-  119  -119  2011/05  [Not refereed][Not invited]
  • EphA2によるShp2のチロシンリン酸化はRAS/MAPK経路の持続的活性化に寄与する(Tyrosine phosphorylation of Shp2 by EphA2 contribute to sustained activation of Ras/MAPK pathway)
    三浦 浩一, 若山 勇紀, 佐邊 壽孝, 望月 直樹  日本細胞生物学会大会講演要旨集  63回-  120  -120  2011/05  [Not refereed][Not invited]
  • TGFβ1による癌的EMTにおけるGEP100-Arf6-AMAP1シグナルの機能解析(HGFR/c-Met-mediated activation of GEP100-Arf6-AMAP1 pathway is an integral part for TGFβ-induced cancerous EMT and invasiveness)
    橋本 あり, 橋本 茂, 大塚 勇太郎, 佐藤 宏紀, 杉野 弘和, 吉河 歩, 梅本 勉, 小野寺 康仁, 福田 諭, 佐邊 壽孝  日本細胞生物学会大会講演要旨集  63回-  151  -151  2011/05  [Not refereed][Not invited]
  • ephrinA1-EphA2シグナルによるezrinの不活性化とMDCK細胞の形態制御(Inactivation of Ezrin by ephrinA1-EphA2 signaling via RhoA contributes to compaction and polarization of MDCK cells)
    若山 勇紀, 三浦 浩一, 佐邊 壽孝, 望月 直樹  日本細胞生物学会大会講演要旨集  63回-  155  -155  2011/05  [Not refereed][Not invited]
  • TGFβ1はGEP100-Arf6-AMAP1経路の活性化によりEMTを誘導し、この活性化は癌幹細胞性と関連する(TGFβ1 activates GEP100-Arf6-AMAP1 pathway to induce EMT, and possible relationship of this activation to cancer stemness)
    橋本 あり, 平野 真理子, 谷野 美智枝, 梅本 勉, 小野寺 康仁, 佐藤 宏紀, 木下 留美子, 南 ジンミン, 大塚 勇太郎, 福田 諭, 白土 博樹, 相沢 慎一, 橋本 茂, 田中 伸哉, 佐邊 壽孝  日本生化学会大会・日本分子生物学会年会合同大会講演要旨集  83回・33回-  2P  -0237  2010/12  [Not refereed][Not invited]
  • 過剰発現したHer2/Neu/ErbB2とGEP100/BRAG2の連係は肺腺癌の自律的な浸潤活性を誘導し、転移を予測するバイオマーカーを提供する(Engagement of GEP100/BRAG2 with overexpressed Her2/Neu/ErbB2 induces autonomous invasive activities and provides a biomarker predictive for metastases of lung adenocarcinoma)
    毛受 暁史, 橋本 茂, 橋本 あり, 伊達 洋至, 佐邊 壽孝  日本生化学会大会・日本分子生物学会年会合同大会講演要旨集  83回・33回-  2P  -0238  2010/12  [Not refereed][Not invited]
  • 癌の悪性化における小胞輸送と糖代謝の役割(Intracellular trafficking and glucose metabolism in tumor malignancy)
    小野寺 康仁, 佐邊 壽孝  日本生化学会大会・日本分子生物学会年会合同大会講演要旨集  83回・33回-  3P  -1019  2010/12  [Not refereed][Not invited]
  • 橋本茂, 橋本あり, 小野寺康仁, 梅本勉, 佐藤宏紀, 毛受暁史, 伊達洋至, 福田諭, 佐邊壽孝  生化学  83回・33回-  ROMBUNNO.3T4-10  -10  2010/12  [Not refereed][Not invited]
  • 癌の悪性化における小胞輸送と糖代謝の役割(Roles of intracellular trafficking and glucose metabolism in tumor malignancy)
    小野寺 康仁, 佐邊 壽孝  日本癌学会総会記事  69回-  83  -83  2010/08  [Not refereed][Not invited]
  • 上皮間葉転換 Arf6-AMAP1経路は癌的EMTに寄与する(EMT (Epithelial Mesenchymal Transition) The Arf6-AMAP1 pathway contributes to cancerous EMT)
    佐邊 壽孝, 小野寺 康仁, 橋本 あり, 橋本 茂  日本癌学会総会記事  69回-  240  -240  2010/08  [Not refereed][Not invited]
  • HER2はGEP100を介して肺癌の浸潤転移を促進する(ErbB2/Her2/Neu employs GEP100 to promote lung cancer invasion and metastasis)
    毛受 暁史, 橋本 茂, 橋本 あり, 伊達 洋至, 佐邊 壽孝  日本癌学会総会記事  69回-  266  -266  2010/08  [Not refereed][Not invited]
  • 低酸素下の癌細胞の浸潤形質獲得とArf6活性化との関連(Hypoxia-induced invasive activity of breast cancer cells involves Arf6 activation)
    橋本 茂, 橋本 あり, 小野寺 康仁, 梅本 勉, 佐藤 宏紀, 毛受 暁史, 佐邊 壽孝  日本癌学会総会記事  69回-  418  -419  2010/08  [Not refereed][Not invited]
  • TGFβによって誘導される癌的EMTにおけるGEP100-Arf6-AMAP1シグナルとHGFRとの相互作用(HGFR-mediated GEP100-Arf6-AMAP1 pathway is an integral part for TGFβ-induced cancerous EMT and invasiveness)
    橋本 あり, 平野 真理子, 梅本 勉, 小野寺 康仁, 佐藤 宏紀, 大塚 勇太郎, 橋本 茂, 佐邊 壽孝  日本癌学会総会記事  69回-  419  -419  2010/08  [Not refereed][Not invited]
  • がん増殖を解くシグナル伝達研究の新展開 癌の悪性化における小胞輸送と糖代謝の役割(New frontiers of signal transduction toward understanding cancer growth Intracellular traffic and glucose metabolism in tumor malignancy)
    小野寺 康仁, 佐邊 壽孝  日本細胞生物学会大会講演要旨集  62回-  105  -105  2010/05  [Not refereed][Not invited]
  • シグナル伝達 EphA2によるリガンド非依存的なRas/MAPK経路の活性化機構(Signal transduction Ligand-independent role of EphA2 in activation of the Ras/MAPK pathway)
    三浦 浩一, 若山 勇紀, 佐邊 壽孝, 望月 直樹  日本細胞生物学会大会講演要旨集  62回-  113  -113  2010/05  [Not refereed][Not invited]
  • 細胞運動・ECM・細胞接着 Zyxinは上皮間葉転換においてアクチン線維再編成に関与し心内膜床の形態形成に寄与する(Cell motility, ECM and cell adhesion Zyxin Mediates Actin Fiber Reorganization in Epithelial-Mesenchymal Transition and Contributes to Endocardial Morphogenesis)
    森 雅樹, 中神 啓徳, 鯉渕 信孝, 三浦 浩一, 佐邊 壽孝, 望月 直樹, 金田 安史  日本細胞生物学会大会講演要旨集  62回-  120  -120  2010/05  [Not refereed][Not invited]
  • 好中球においてゴルジ体に局在するArfGEFのPI3Kγによる局在変化と活性化は、GPCR刺激と細胞運動を結びつけている(Translocation and activation of a Golgi-localizing ArfGEF via PI3Kγ links GPCR stimulation with directional migration in neutrophils)
    真崎 雄一, 佐邊 壽孝  日本細胞生物学会大会講演要旨集  62回-  138  -138  2010/05  [Not refereed][Not invited]
  • Arf6を介したephrinA1-EphA2シグナルによるEzrinの不活性化はMDCK細胞の形態変化に寄与する(Inactivation of Ezrin by ephrinA1-EphA2 signaling via Arf6 contributes to compaction of MDCK cells)
    若山 勇紀, 三浦 浩一, 佐邊 壽孝, 望月 直樹  日本細胞生物学会大会講演要旨集  62回-  141  -141  2010/05  [Not refereed][Not invited]
  • GEP100-Arf6-AMAP1経路は癌浸潤と血管新生に対する共通の分子標的である(GEP100-Arf6-AMAP1 pathway is activated by VEGFR2 and promotes vascular remodeling and VE-cadherin endocytosis in endothelial cells)
    橋本 あり, 橋本 茂, 小川 栄治, 毛受 暁史, 森重 真毅, 佐邊 壽孝  日本細胞生物学会大会講演要旨集  62回-  161  -161  2010/05  [Not refereed][Not invited]
  • 癌の転移部における休止と再発 EMTから考える
    佐邊 壽孝  日本細胞生物学会大会講演要旨集  62回-  215  -215  2010/05  [Not refereed][Not invited]
  • 佐邊壽孝  がん研究に係わる特定領域研究研究報告集録 平成21年度  197  2010  [Not refereed][Not invited]
  • 佐邊壽孝  生化学  ROMBUNNO.3S1A-2  2009/09/25  [Not refereed][Not invited]
  • 低分子量GTPaseから眺めるメンブレントラフィック研究の新展開 メンブレントラフィックと癌浸潤形質獲得機序
    佐邊 壽孝  日本生化学会大会プログラム・講演要旨集  82回-  3S1a  -2  2009/09  [Not refereed][Not invited]
  • EphA2受容体によるArf6の不活性化と上皮細胞間の接着制御(EphA2 engages Git1 to suppress Arf6 activity and modulate epithelial cell-cell contacts)
    三浦 浩一, Nam Jin-Min, 児島 千恵, 望月 直樹, 佐邊 壽孝  日本生化学会大会プログラム・講演要旨集  82回-  2T15p  -10  2009/09  [Not refereed][Not invited]
  • 乳癌細胞における恒常的なNF-kappaBの活性化に対するNIKおよびA20の役割(Roles for NIK and A20 in constitutive NF-kappaB activation in breast cancer cells)
    斉藤 愛記, 橋本 茂, 佐邊 壽孝, 山岡 昇司  日本癌学会総会記事  68回-  323  -323  2009/08  [Not refereed][Not invited]
  • セマフォリンシグナル伝達とエフリンシグナル伝達に関する最新の知見 上皮細胞間接触を制御するArf6のGit1による抑制におけるEphA2の関与(Recent advances in semaphorin signaling and ephrin signaling EphA2 engages Git1 to suppress Arf6 activity modulating epithelial cell-cell contacts)
    三浦 浩一, Nam Jin-Min, 児島 千恵, 望月 直樹, 佐邊 壽孝  日本細胞生物学会大会講演要旨集  61回-  112  -112  2009/05  [Not refereed][Not invited]
  • 好中球のケモタキシスにおけるGBF1の役割(Roles of GBF1 in neutrophil chemotaxis)
    真崎 雄一, 佐邊 壽孝  日本細胞生物学会大会講演要旨集  61回-  148  -148  2009/05  [Not refereed][Not invited]
  • 佐邊壽孝  がん研究に係わる特定領域研究研究報告集録 平成20年度  201  2009  [Not refereed][Not invited]
  • 佐邊壽孝  がん特定研究5領域合同シンポジウムプログラム・抄録集 平成20年度  12  2009  [Not refereed][Not invited]
  • 蛋白質のリン酸化と細胞応答 GEP100-Arf6-AMAP1-cortactinシグナル経路は癌浸潤と血管新生に共通である
    橋本 あり, 橋本 茂, 小川 栄治, 廣瀬 まゆみ, 森重 真毅, 毛受 暁史, 小野寺 康仁, 渋谷 正史, 佐邊 壽孝  日本生化学会大会・日本分子生物学会年会合同大会講演要旨集  81回・31回-  4S2  -3  2008/11  [Not refereed][Not invited]
  • 濃度勾配性因子による細胞極性・細胞運動の制御 組織構築、創傷治癒およびがんの浸潤・転移の機構解明を目指して 乳癌浸潤転移におけるEGFR-GEP100-Arf6-AMAP1経路 微小環境との相互作用
    佐邊 壽孝, 橋本 茂, 森重 真毅, 橋本 あり, 小川 栄二, 矢野 元, Nam Jinmin, 小野寺 康仁  日本生化学会大会・日本分子生物学会年会合同大会講演要旨集  81回・31回-  4S14  -3  2008/11  [Not refereed][Not invited]
  • 橋本茂, 森重真毅, 小川栄治, 橋本あり, 小野寺康仁, 佐邊壽孝  実験医学  26-  (15)  2349  -2355  2008/09/15  [Not refereed][Not invited]
  • 【シグナル伝達研究 2008'09 疾患発症の分子メカニズムと実現化する分子標的薬開発】 シグナル伝達研究 因子から現象へ がんの浸潤形質獲得過程における低分子量Gタンパク質Arf6シグナル伝達
    橋本 茂, 森重 真毅, 小川 栄治, 橋本 あり, 小野寺 康仁, 佐邊 壽孝  実験医学  26-  (15)  2349  -2355  2008/09  [Not refereed][Not invited]
     
    がんの主たる脅威は、浸潤・転移性の獲得にある。恒常性を維持した細胞浸潤は、胎児期の発生・分化過程や創傷治癒、免疫応答などでみられるが、どのような制御機構の破綻ががんの浸潤形質を誘導しているのか未だ不明のままである。がんの浸潤形質獲得は、がん細胞自身のgenetic/epigeneticな変異の多段階的蓄積のみならず、微小環境の変化に依存している。われわれは、低分子量Gタンパク質Arf6を中心としたシグナル伝達経路の活性化が浸潤形質の獲得に必須であることを見出した。本稿では、がん浸潤形質の誘導に関連した最近の知見を概説するとともに、われわれのモデルについて紹介したい。(著者抄録)
  • 低酸素環境とEMTにおける乳腺上皮細胞の浸潤性獲得に関連する網羅的解析(Comprehensive analysis for the acquisition of invasiveness of mammary epithelial cells under hypoxia and EMT)
    橋本 茂, 橋本 あり, 魏 樹梅, 三浦 浩一, 毛受 暁史, 佐邊 壽孝  日本癌学会総会記事  67回-  101  -101  2008/09  [Not refereed][Not invited]
  • GEP100-Arf6経路は血管新生と癌浸潤に共通のシグナル経路である(Common usage of the GEP100-Arf6 signaling pathway in tumor invasion, angiogenesis, and vascular permeability)
    橋本 あり, 橋本 茂, 小川 栄治, 廣瀬 まゆみ, 森重 真毅, 毛受 暁史, 渋谷 正史, 佐邊 壽孝  日本癌学会総会記事  67回-  296  -296  2008/09  [Not refereed][Not invited]
  • 癌浸潤転移における細胞運動のメカニズム 血管新生と癌浸潤に共通なシグナル経路(Molecular mechanisms of cell migration in cancer invasion and metastasis Common usage of an Arf6-GEP100 signaling pathway in angiogenesis and tumor invasion)
    橋本 あり, 橋本 茂, 小川 栄治, 廣瀬 まゆみ, 高島 成二, 森重 真毅, 毛受 暁史, 南 ジンミン, 真崎 雄一, 北風 政史, 渋谷 正史, 佐邊 壽孝  日本細胞生物学会大会講演要旨集  60回-  95  -95  2008/06  [Not refereed][Not invited]
  • Tobias Bauer, Nami Motosugi, Koichi Miura, Hisataka Sabe, Takashi Hiiragi  GENESIS  46-  (3)  152  -162  2008/03  [Not refereed][Not invited]
     
    Cytokinesis is a complex process that involves dynamic cortical rearrangement. Our recent time-lapse recordings of the mouse egg unexpectedly revealed a high motility of the second polar body (2pb). Experiments to address its underlying mechanism show that neither mechanical compression by the zona pellucida nor the connection via the mid-body is required for the 2pb movement. Time-lapse recordings establish that the 2pb moves together with the cell membrane. These recordings, in which cell surface proteins are labeled with fluorescent latex-microbeads or monovalent antibodies against whole mouse proteins, indicate that the majority of the surface proteins dynamically accumulate in the cleavage furrow at every cell division. Comparable dynamics of the cell surface proteins, and specifically of E-cadherin, are also observed in cultured epithelial cells. The surface protein dynamics are closely correlated with, and dependent on, those of the underlying cortical actin. The cortical actin network may form a scaffold for membrane proteins and thereby transfer them during contractile ring formation toward the cleavage furrow. Immobilization of surface proteins by tetravalent lectin-mediated crosslinking results in the failure of cleavage, demonstrating that the observed protein dynamics are essential for cytokinesis. We propose that dynamic rearrangement of the cell surface proteins is a common feature of cytokinesis, playing a key role in modifying the mechanical properties of the cell membrane during cortical ingression.
  • Hajime Yano, Itaru Kobayashi, Yasuhito Onodera, Frederic Luton, Michel Franco, Yuichi Mazaki, Shigeru Hashimoto, Kazuhiro Iwai, Ze'ev Ronai, Hisataka Sabe  MOLECULAR BIOLOGY OF THE CELL  19-  (3)  822  -832  2008/03  [Not refereed][Not invited]
     
    The small GTP-binding protein Arf6 regulates membrane remodeling at cell peripheries and plays crucial roles in higher orders of cellular functions including tumor invasion. Here we show that Fbx8, an F-box protein bearing the Sec7 domain, mediates ubiquitination of Arf6. This ubiquitination did not appear to be linked to immediate proteasomal degradation of Arf6, whereas Fbx8 knockdown caused hyperactivation of Arf6. Expression of Fbx8 protein was substantially lost in several breast tumor cell lines, in which Arf6 activity is pivotal for their invasion. Forced expression of Fbx8 in these cells suppressed their Arf6 activities and invasive activities, in which the F-box and Sec7 domains of Fbx8 are required. Together with the possible mechanism as to how Fbx8-mediated ubiquitination interferes with the functions of Arf6, we propose that Fbx8 provides a novel suppressive control of Arf6 activity through noncanonical ubiquitination. Our results indicate that dysfunction of Fbx8 expression may contribute to the invasiveness of some breast cancer cells.
  • S. Kon, K. Tanabe, T. Watanabe, H. Sabe, M. Satake  Exp Cell Res.  314-  (7)  1415  -1428  2008  [Not refereed][Not invited]
  • M. Morishige, S. Hashimoto, E. Ogawa, S. Wei, A. Hashimoto, A. Yamada, Y. Mazaki, Y. Toda, H. Wada, H. Kobayashi, H. Sabe  Nat. Cell Biol  10-  (1)  85  -92  2008  [Not refereed][Not invited]
  • 橋本あり, 橋本茂, 小川栄治, 廣瀬まゆみ, 森重真毅, 毛受暁史, 小野寺康仁, 渋谷正史, 佐邊壽孝  生化学  4S2-3  2008  [Not refereed][Not invited]
  • 佐邊壽孝, 橋本茂, 森重真毅, 橋本あり, 小川栄二, 矢野元, NAM Jinmin, 小野寺康仁  生化学  4S14-3  2008  [Not refereed][Not invited]
  • 橋本茂, 三浦浩一, 橋本あり, WEI Shumei, 毛受暁史, 佐邊壽孝  生化学  2P-0426  2008  [Not refereed][Not invited]
  • 佐邊壽孝  がん研究に係わる特定領域研究研究報告集録 平成19年度  216  2008  [Not refereed][Not invited]
  • Fbx8によるユビキチン化を介するArf6の抑制的制御と上皮組織形態形成との関連の可能性について(Fbx8 makes Arf6 refractory to function via ubiquitination: implication in epithelial tissue organization)
    矢野 元, 小野寺 康仁, 鳥井 郁子, 真崎 雄一, 橋本 茂, 辻村 亨, 佐邊 壽孝  日本生化学会大会・日本分子生物学会年会合同大会講演要旨集  80回・30回-  1T21  -3  2007/11  [Not refereed][Not invited]
  • 真崎雄一, 佐邊壽孝  生化学  80回・30回-  2P-0421  -15  2007/11  [Not refereed][Not invited]
  • 森重真毅, 橋本茂, 阿部竜也, 藤木稔, 古林秀則, 佐邊壽孝  日本脳神経外科学会総会抄録集(CD-ROM)  66th-  2K-P42-12-3  -P42  2007/10  [Not refereed][Not invited]
  • EGFレセプターシグナルを介した乳癌細胞の浸潤形質誘導における必須因子Arf6の活性化機序(GEP100 links epidermal growth factor receptor signaling to Arf6 activation to induce breast cancer invasion)
    橋本 茂, 森重 真毅, 小川 栄治, 廣瀬 まゆみ, 魏 樹梅, 橋本 あり, 山田 敦子, 矢野 元, 仁尾 義則, 和田 洋巳, 古林 秀則, 佐邊 壽孝  日本癌学会総会記事  66回-  58  -58  2007/08  [Not refereed][Not invited]
  • Git1は、Arf6活性化E-cadherin誘発細胞接着を抑制するために、Nckを介してリガンド活性化EphA2と結合する(Git1 links to ligand-activated EphA2 via Nck to suppress Arf6 activity modulatinq E-cadherin-mediated cell-cell contacts)
    三浦 浩一, Nam Jin-Min, 佐邊 壽孝  日本癌学会総会記事  66回-  132  -132  2007/08  [Not refereed][Not invited]
  • 癌浸潤及び血管新生におけるArf6活性化の意義(Common usage of Arf6-signaling pathway in tumor invasion and angiogenesis)
    橋本 あり, 南 ジンミン, 森重 真毅, 毛受 暁史, 橋本 茂, 渋谷 正史, 佐邊 壽孝  日本癌学会総会記事  66回-  142  -142  2007/08  [Not refereed][Not invited]
  • Arf6をユビキチン化するE3リガーゼFbx8のがんにおける発現不全(Loss of Fbx8, a component of E3 ligase mediating Arf6 ubiquitination, in different human tumors)
    矢野 元, 真崎 雄一, 橋本 茂, 辻村 亨, 佐邊 壽孝  日本癌学会総会記事  66回-  142  -143  2007/08  [Not refereed][Not invited]
  • EMT進行と細胞移動における赤血球タンパク質band4.1-like5(Ebl5)の重要な役割(Erythrocyte protein band4.1-like5 (Ebl5) plays an essential role in the EMT progression and cell migration)
    平野 真理子, 佐邊 壽孝, 相澤 慎一  日本発生生物学会・日本細胞生物学会合同大会要旨集  40回・59回-  169  -169  2007/05  [Not refereed][Not invited]
  • Jin-Min Nam, Yasuhito Onodera, Yuichi Mazaki, Hiroyuki Miyoshi, Shigeru Hashimoto, Hisataka Sabe  EMBO JOURNAL  26-  (3)  647  -656  2007/02  [Not refereed][Not invited]
     
    Expression of AMAP1 correlates well with the invasive phenotypes and malignancy of human primary breast carcinomas. AMAP1 recruits its binding proteins, such as cortactin and paxillin, to sites of Arf6 activation to form invadopodia. A mouse ortholog of AMAP1, ASAP1, is known to bind to CIN85, a binding partner of an E3 ligase, Cbl. Here, we found that CIN85 colocalizes with AMAP1 at invadopodia, and binding of AMAP1 with CIN85 is important for the invasive activities of breast cancer cells, including MDA-MB-231. siRNA-mediated silencing of CIN85, as well as Cbl, also inhibited the invasion. We moreover found that AMAP1 is monoubiquitinated, rather than polyubiquitinated, by virtue of Cbl and provide evidence that the ability of AMAP1 to be monoubiquitinated is important for its involvement in invasion. Our results indicate that CIN85, as well as Cbl, which is a well-known suppressor of growth factor receptor signaling, can be positively involved in tumor invasion, and suggest that a complex epigenetic process is involved in AMAP1 function in breast cancer cell invasion.
  • CIN85, a Cbl-interacting multiadaptor protein, is a component of AMAP1-mediated breast cancer invasion machinery.
    J. Nam, Y. Onodera, Y. Mazaki, H. Miyoshi, S. Hashimoto, H. Sabe  Nat. Immunol.  7-  724  -731  2007  [Not refereed][Not invited]
  • 平野真理子, 佐邊壽孝, 相澤慎一  生化学  3P-1004  2007  [Not refereed][Not invited]
  • 佐邊壽孝  がん研究に係わる特定領域研究研究報告集録 平成18年度  216  2007  [Not refereed][Not invited]
  • 宮田真理子, ANTONIS Koromilas, 佐邊壽孝  生化学  3P-0641  2007  [Not refereed][Not invited]
  • Eph受容体によるE-カドヘリンを介した細胞間接着の制御
    三浦 浩一, 佐邊 壽孝  日本癌学会総会記事  65回-  241  -241  2006/09  [Not refereed][Not invited]
  • 乳癌細胞の浸潤におけるAMAP1のユビキチン化の役割(Property of AMAP1 to be monoubiquitinated via binding with CIN85 and Cbl is crucial for breast cancer invasive activity)
    Nam Jin-Min, 小野寺 康仁, 橋本 茂, 佐邊 壽孝  日本癌学会総会記事  65回-  295  -296  2006/09  [Not refereed][Not invited]
  • 乳癌細胞におけるFbox8発現不全と浸潤性獲得の関連性
    矢野 元, 橋本 茂, 佐邊 壽孝  日本癌学会総会記事  65回-  296  -296  2006/09  [Not refereed][Not invited]
  • AMAP1/コータクチン複合体形成阻害による血管新生及びmesenchymal型とamoeboid型癌浸潤の阻害
    橋本 あり, 山田 敦子, 森重 真毅, 橋本 茂, 佐邊 壽孝  日本癌学会総会記事  65回-  311  -311  2006/09  [Not refereed][Not invited]
  • 乳癌細胞の浸潤形質獲得における必須因子Arf6の活性化機序
    橋本 茂, 森重 真毅, 古林 秀則, 橋本 あり, 魏 樹梅, 山田 敦子, 佐邊 壽孝  日本癌学会総会記事  65回-  456  -456  2006/09  [Not refereed][Not invited]
  • 三浦浩一, 佐邊壽孝  日本癌学会学術総会記事  65th-  241  2006/08/28  [Not refereed][Not invited]
  • 矢野元, 橋本茂, 佐邊壽孝  日本癌学会学術総会記事  65th-  296  2006/08/28  [Not refereed][Not invited]
  • 橋本茂, 森重真毅, 古林秀則, 橋本あり, WEI Shumei, 山田敦子, 佐邊壽孝  日本癌学会学術総会記事  65th-  456  2006/08/28  [Not refereed][Not invited]
  • 橋本あり, 山田敦子, 森重真毅, 橋本茂, 佐邊壽孝  日本癌学会学術総会記事  65th-  311  2006/08/28  [Not refereed][Not invited]
  • Y Mazaki, S Hashimoto, T Tsujimura, M Morishige, A Hashimoto, K Aritake, A Yamada, JM Nam, H Kiyonari, K Nakao, H Sabe  NATURE IMMUNOLOGY  7-  (7)  724  -731  2006/07  [Not refereed][Not invited]
     
    In neutrophils, superoxide anion production generally accompanies chemotaxis and functions in killing invading pathogens. The GIT2 GTPase-activating protein binds to the guanine nucleotide-exchange factor alpha PIX. Here we show that GIT2 was necessary for directional chemotaxis and for the suppression of superoxide production in G protein-coupled receptor-stimulated neutrophils. GIT2 was also necessary for the orientation of superoxide production toward chemoattractant sources. GIT2 suppressed the activity of ADP ribosylation factor 1 and was a component of the G beta gamma subunit-mediated direction-sensing machinery 'downstream' of G protein-coupled receptor signaling. This study establishes a function for GIT2 in linking chemotaxis and superoxide production in neutrophils and shows that loss of GIT2 in vivo leads to an immunodeficient state.
  • 小野寺康仁, 佐邊壽孝  実験医学  24-  (10)  1433  -1439  2006/06/20  [Not refereed][Not invited]
  • 【分子メカニズムから解き明かす疾患のサイエンス】 癌 癌細胞の浸潤・転移におけるタンパク質の相互作用と動態の制御
    小野寺 康仁, 佐邊 壽孝  実験医学  24-  (10)  1433  -1439  2006/06  [Not refereed][Not invited]
     
    癌の転移において癌細胞の浸潤活性は最も重要な性質の1つである.癌細胞は,既存分子の相互作用を改変して細胞-細胞間接着や細胞-基質間接着,運動性や細胞外基質分解活性を巧みに制御し,浸潤・転移を行うものと考えられる.これらの過程に特異的に関与するタンパク質間相互作用の同定は,正常組織への副作用を抑えた浸潤・転移阻害薬の開発において有効な情報を与えることが期待される(著者抄録)
  • S Hashimoto, M Hiroso, A Hashimoto, M Morishige, A Yamada, H Hosaka, KI Akagi, E Ogawa, C Oneyama, T Agatsuma, M Okada, H Kobayashi, H Wada, H Nakano, T Ikegami, A Nakagawa, H Sabe  PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA  103-  (18)  7036  -7041  2006/05  [Not refereed][Not invited]
     
    Invasive potentials of carcinomas greatly contribute to their metastasis, which is a major threat in most cancers. We have recently shown that Arf6 plays a pivotal role in breast cancer invasive activities and identified AMAP1 as an effector of GTP-Arf6 in invasion. Expression of AMAP1 correlates well with invasive phenotypes of primary tumors of the human breast. We also have shown that AMAP1 functions by forming a trimeric protein complex with cortactin and paxillin. In this complex, AMAP1 binds to the src homology 3 (SH3) domain of cortactin via its proline-rich peptide, SKKRPPPPPPGHKRT. SH3 domains are known to bind generally to the proline-rich ligands with a one-to-one stoichiometry. We found that AMAP1/cortactin binding is very atypical in its stoichiometry and interface structure, in which one AMAP1 proline-rich peptide binds to two cortactin SH3 domains simultaneously. We made a cell-permeable peptide derived from the AMAP1 peptide, and we show that this peptide specifically blocks AMAP1/cortactin binding, but not other canonical SH3/proline bindings, and effectively inhibits breast cancer invasion and metastasis. Moreover, this peptide was found to block invasion of other types of cancers, such as glioblastomas and lung carcinomas. We also found that a small-molecule compound, UCS15A, which was previously judged as a weak inhibitor against canonical SH3/proline bindings, effectively inhibits AMAP1/cortactin binding and breast cancer invasion and metastasis. Together with fine structural analysis, we propose that the AMAP1/cortactin complex, which is not detected in normal mammary epithelial cells, is an excellent drug target for cancer therapeutics.
  • 廣瀬まゆみ, 橋本茂, 橋本あり, 森重真毅, 山田敦子, 保坂晴美, 赤木謙一, 小川栄治, 池上貴久, 中川敦史, 佐邊壽孝  日本蛋白質科学会年会プログラム・要旨集  6th-  67  2006/03/31  [Not refereed][Not invited]
  • 佐邊壽孝  Arf6による細胞運動性制御の分子機構の解析 平成16-17年度 No.16370090  97P  2006  [Not refereed][Not invited]
  • ヒト単球芽様細胞U937の分化におけるAMAP1蛋白質量の上昇機構の解析
    宮田 真理子, 佐邊 壽孝  日本免疫学会総会・学術集会記録  35-  54  -54  2005/11  [Not refereed][Not invited]
  • 細胞間接触によるArf6の負の制御
    三浦 浩一, 小野寺 康仁, 佐邊 壽孝  日本癌学会総会記事  64回-  167  -167  2005/09  [Not refereed][Not invited]
  • Arf6ユビキチン化機構と乳癌細胞浸潤性獲得過程との関連性
    矢野 元, 橋本 茂, 小野寺 康仁, 佐邊 壽孝  日本癌学会総会記事  64回-  173  -173  2005/09  [Not refereed][Not invited]
  • 浸潤性乳癌細胞のAMAP1/コータクチン相互作用インターフェースの癌浸潤阻害剤の標的としての評価
    橋本 茂, 廣瀬 まゆみ, 橋本 あり, 森重 真毅, 山田 敦子, 小野寺 康仁, 小川 栄治, 和田 洋巳, 池上 貴久, 中川 敦史, 佐邊 壽孝  日本癌学会総会記事  64回-  309  -309  2005/09  [Not refereed][Not invited]
  • 浸潤・転移・血管新生研究の進歩 乳癌における浸潤形質獲得過程
    佐邊 壽孝, 小野寺 康仁, ナム・ジンミン, 橋本 あり, 森重 真毅, 橋本 茂  日本癌学会総会記事  64回-  513  -513  2005/09  [Not refereed][Not invited]
  • AJ. Woods, T. Kantidakis, H. Sabe, DR. Critchley & JC. Norman. Interaction of paxillin with poly(A)-binding protein 1 and its role in focal adhesion turnover and cell migration.
    Mol.Cell Biol.  25:3763-3773.-  2005  [Not refereed][Not invited]
     
    2005.
  • EMBO J.  24:963-973.-  2005  [Not refereed][Not invited]
     
    Y. Onodera, S. Hashimoto, A. Hashimoto, M. Morishige, Y. Mazaki, A. Yamada, E. Ogawa, M. Adachi, T. Sakurai, T. Manabe, H. Wada, N. Matsuura & H. Sabe.
    Expression of AMAP1, an ArfGAP, provides novel targets to inhibit breast cancer invasive activities.

    2005.
  • AJ. Woods, T. Kantidakis, H. Sabe, DR. Critchley & JC. Norman. Interaction of paxillin with poly(A)-binding protein 1 and its role in focal adhesion turnover and cell migration.
    Mol.Cell Biol.  25:3763-3773.-  2005  [Not refereed][Not invited]
     
    2005.
  • EMBO J.  24:963-973.-  2005  [Not refereed][Not invited]
     
    Y. Onodera, S. Hashimoto, A. Hashimoto, M. Morishige, Y. Mazaki, A. Yamada, E. Ogawa, M. Adachi, T. Sakurai, T. Manabe, H. Wada, N. Matsuura & H. Sabe.
    Expression of AMAP1, an ArfGAP, provides novel targets to inhibit breast cancer invasive activities.

    2005.
  • 【発生・分化再生研究2005】 形態形成 細胞間相互作用とシグナル伝達 上皮細胞における接着分子機能の動的制御
    三浦 浩一, 佐邊 壽孝  実験医学  23-  (1)  46  -51  2005/01  [Not refereed][Not invited]
  • 乳癌の浸潤活性におけるArf6の要求性
    佐邊 壽孝  上原記念生命科学財団研究報告集  18-  203  -205  2004/11  [Not refereed][Not invited]
     
    低分子量G蛋白質Arf6(ADP-ribosylation factor 6)活性とその発現レベルと乳癌浸潤性との関係について検討した.MDA-MB-231細胞をはじめとする種々のヒト乳癌細胞株は全てATCCより入手した.siRNAは常法により施行した.Arf6は乳癌におけるinvadopodiaの一要素で,siRNA法によってArf6の発現を抑制すると乳癌の浸潤活性を効率良く阻害できた.mRNAは培養された正常乳腺上皮細胞でもすでに発現し,浸潤性とは相関していなかった.種々のcDNAや方法論を用いて解析したところ,Arf6の持続的活性化が乳癌の浸潤活性に必須であることが示唆された
  • がんの微小環境制御による浸潤・転移治療を目指して 乳癌の浸潤性獲得 隠された浸潤装置
    佐邊 壽孝  日本癌学会総会記事  63回-  391  -391  2004/09  [Not refereed][Not invited]
  • 乳癌細胞におけるArf6蛋白質の量的制御機構と浸潤性獲得
    矢野 元, 古林 格, 小野寺 康仁, 橋本 茂, 佐邊 壽孝  日本癌学会総会記事  63回-  265  -265  2004/09  [Not refereed][Not invited]
  • Arf6は乳癌細胞の浸潤性獲得において必須の分子装置の一つである
    橋本 茂, 小野寺 康仁, 橋本 あり, 森重 真毅, 田中 美和, 浜口 道成, 佐邊 壽孝  日本癌学会総会記事  63回-  265  -265  2004/09  [Not refereed][Not invited]
  • 浸潤性乳癌細胞におけるAMAP1の発現と機能
    小野寺 康仁, 橋本 茂, 真崎 雄一, 橋本 あり, 森重 真毅, 松浦 成昭, 佐邊 壽孝  日本癌学会総会記事  63回-  267  -267  2004/09  [Not refereed][Not invited]
  • Kojima C, Hashimoto A, Yabuta I, Hirose M, Hashimoto S, Kanaho Y, Sumimoto H, Ikegami T, Sabe H. Regulation of Bin1 SH3 domain binding by phosphoinositides.
    EMBO J.  2004  [Not refereed][Not invited]
  • Yano H, Mazaki Y, Kurokawa K, Hanks SK, Matsuda M, Sabe H. Roles played by a subset of integrin signaling molecules in cadherin-based cell-cell adhesion.
    J Cell Biol.  166(2):283-95.-  2004  [Not refereed][Not invited]
  • Romanova LY, Hashimoto S, Chay KO, Blagosklonny MV, Sabe H, Mushinski JF. Phosphorylation of paxillin tyrosines 31 and 118 controls polarization and motility of lymphoid cells and is PMA-sensitive.
    J Cell Sci.  117(Pt 17):3759-68.-  2004  [Not refereed][Not invited]
  • Hashimoto S, Onodera Y, Hashimoto A, Tanaka M, Hamaguchi M, Yamada A, Sabe H. Requirement for Arf6 in breast cancer invasive activities.
    Proc Natl Acad Sci U S A.  101(17):6647-52.-  2004  [Not refereed][Not invited]
  • Kojima C, Hashimoto A, Yabuta I, Hirose M, Hashimoto S, Kanaho Y, Sumimoto H, Ikegami T, Sabe H. Regulation of Bin1 SH3 domain binding by phosphoinositides.
    EMBO J.  2004  [Not refereed][Not invited]
  • Yano H, Mazaki Y, Kurokawa K, Hanks SK, Matsuda M, Sabe H. Roles played by a subset of integrin signaling molecules in cadherin-based cell-cell adhesion.
    J Cell Biol.  166(2):283-95.-  2004  [Not refereed][Not invited]
  • Romanova LY, Hashimoto S, Chay KO, Blagosklonny MV, Sabe H, Mushinski JF. Phosphorylation of paxillin tyrosines 31 and 118 controls polarization and motility of lymphoid cells and is PMA-sensitive.
    J Cell Sci.  117(Pt 17):3759-68.-  2004  [Not refereed][Not invited]
  • Hashimoto S, Onodera Y, Hashimoto A, Tanaka M, Hamaguchi M, Yamada A, Sabe H. Requirement for Arf6 in breast cancer invasive activities.
    Proc Natl Acad Sci U S A.  101(17):6647-52.-  2004  [Not refereed][Not invited]
  • 皮膚線維芽細胞におけるUltraviolet A照射によるマトリックスメタロプロテアーゼ1の発現はMacrophage migration inhibitory factorを介する
    渡辺 宏数, 清水 忠道, 西平 順, 阿部 理一郎, 中山 敏則, 谷口 克, 佐邊 壽孝, 石橋 輝雄, 清水 宏  生化学  75-  (12)  1570  -1570  2003/12  [Not refereed][Not invited]
  • 医学・医療の進歩を世界へ向けて 細胞内情報伝達・分子細胞医学 細胞の接着・形態形成と細胞内情報伝達 Endosomal Recyclingと細胞運動性制御
    佐邊 壽孝  日本医学会総会会誌  26回-  (2)  210  -210  2003/12  [Not refereed][Not invited]
  • カドヘリン接着形成におけるインテグリンシグナル分子群の役割
    矢野 元, 真崎 雄一, 三浦 浩一, 佐邊 壽孝  日本癌学会総会記事  62回-  40  -40  2003/08  [Not refereed][Not invited]
  • J Biochem.  134(4):485-9-  2003  [Not refereed][Not invited]
  • Watanabe H, Shimizu T, Nishihira J, Abe R, Nakayama T, Taniguchi M, Sabe H, Ishibashi T, Shimizu H. Ultraviolet A-induced production of matrix metalloproteinase-1 is mediated by macrophage migration inhibitory factor (MIF) in human dermal fibroblasts.
    J Biol Chem.  279(3):1676-83-  2003  [Not refereed][Not invited]
  • DIDIER C, BRODAY L, BHOUMIK A, ISRAELI S, THOMAS S M, TURNER C E, HENDERSON S, SABE H, RONAI Z  Mol. Cell Biol.  23-  (15)  5331  -5345  2003  [Not refereed][Not invited]
     
    C. Didier, L. Broday, A. Bhoumik, S. Israeli, S. Takahashi, K. Nakayama, S. M. Thomas, C. E. Turner, S. Henderson, H. Sabe. & Z. Ronai.
    RNF5 - a RING finger protein that regulates cell motility by targeting paxillin ubiquitination and altered localization.
  • J Biochem.  134(4):485-9-  2003  [Not refereed][Not invited]
  • Watanabe H, Shimizu T, Nishihira J, Abe R, Nakayama T, Taniguchi M, Sabe H, Ishibashi T, Shimizu H. Ultraviolet A-induced production of matrix metalloproteinase-1 is mediated by macrophage migration inhibitory factor (MIF) in human dermal fibroblasts.
    J Biol Chem.  279(3):1676-83-  2003  [Not refereed][Not invited]
  • Mol. Cell Biol.  23-  (15)  5331  -5345  2003  [Not refereed][Not invited]
     
    C. Didier, L. Broday, A. Bhoumik, S. Israeli, S. Takahashi, K. Nakayama, S. M. Thomas, C. E. Turner, S. Henderson, H. Sabe. & Z. Ronai.
    RNF5 - a RING finger protein that regulates cell motility by targeting paxillin ubiquitination and altered localization.
  • SABE Hisataka  生化学  74-  (12)  1429  -1440  2002/12/25  [Not refereed][Not invited]
     
    細胞運動は細胞の接着,前方部の伸展,核を含めた細胞体の牽引,後方部の脱接着,細胞体の前方への移動の少なくとも五つのレパートリーからなり,それらは細胞内の物理力の発生や運動方向極性の決定機序などとも統制され,時空間的な順序をもっておこる.物理力や極性発生の機構も互いに提携し統制され,その結果,意味のある効率的な運動がなされる.細胞内シグナル伝達,細胞骨格再構成,膜の細胞内輸送や再構成などの各々個別の過程が,これらの機序や機構を作動させる.現在の細胞生物学における重要な問題点の一つはこれら個別の細胞内過程を互いに調整し統制している機構の分子的実態を明らかにすることである
  • A Tsubouchi, J Sakakura, R Yagi, Y Mazaki, E Schaefer, H Yano, H Sabe  JOURNAL OF CELL BIOLOGY  159-  (4)  673  -683  2002/11  [Not refereed][Not invited]
     
    RhoA activity is transiently inhibited at the initial phase of integrin engagement, when Cdc42- and/or Rac1-mediated membrane spreading and ruffling predominantly occur. Paxillin, an integrin-assembly protein, has four major tyrosine phosphorylation sites, and the phosphorylation of Tyr31 and Tyrl 18 correlates with cell adhesion and migration. We found that mutation of Tyr31/118 caused enhanced activation of RhoA and premature formation of stress fibers with substantial loss of efficient membrane spreading and ruffling in adhesion and migration of NMuMG cells. These phenotypes were similar to those induced by RhoA(G14V) in parental cells, and could be abolished by expression of RhoA(T19N), Rac1(G12V), or p190RhoGAP in the mutant-expressing cells. Phosphorylated Tyr31/118 was found to bind to two src homology (SH)2 domains of p120RasGAP, with coprecipitation of endogenous paxillin with p120RasGAP. p190RhoGAP is known to be a major intracellular binding partner for the p120RasGAP SH2 domains. We found that Tyr31/118-phosphorylated paxillin competes with p190RhoGAP for binding to p120RasGAP, and provides evidence that p190RhoGAP freed from p120RasGAP efficiently suppresses RhoA activity during cell adhesion. We conclude that Tyr31/118-phosphorylated paxillin serves as a template for the localized suppression of RhoA activity and is necessary for efficient membrane spreading and ruffling in adhesion and migration of NMuMG cells.
  • A Tsubouchi, J Sakakura, R Yagi, Y Mazaki, E Schaefer, H Yano, H Sabe  JOURNAL OF CELL BIOLOGY  159-  (4)  673  -683  2002/11  [Not refereed][Not invited]
     
    RhoA activity is transiently inhibited at the initial phase of integrin engagement, when Cdc42- and/or Rac1-mediated membrane spreading and ruffling predominantly occur. Paxillin, an integrin-assembly protein, has four major tyrosine phosphorylation sites, and the phosphorylation of Tyr31 and Tyrl 18 correlates with cell adhesion and migration. We found that mutation of Tyr31/118 caused enhanced activation of RhoA and premature formation of stress fibers with substantial loss of efficient membrane spreading and ruffling in adhesion and migration of NMuMG cells. These phenotypes were similar to those induced by RhoA(G14V) in parental cells, and could be abolished by expression of RhoA(T19N), Rac1(G12V), or p190RhoGAP in the mutant-expressing cells. Phosphorylated Tyr31/118 was found to bind to two src homology (SH)2 domains of p120RasGAP, with coprecipitation of endogenous paxillin with p120RasGAP. p190RhoGAP is known to be a major intracellular binding partner for the p120RasGAP SH2 domains. We found that Tyr31/118-phosphorylated paxillin competes with p190RhoGAP for binding to p120RasGAP, and provides evidence that p190RhoGAP freed from p120RasGAP efficiently suppresses RhoA activity during cell adhesion. We conclude that Tyr31/118-phosphorylated paxillin serves as a template for the localized suppression of RhoA activity and is necessary for efficient membrane spreading and ruffling in adhesion and migration of NMuMG cells.
  • A Tsubouchi, J Sakakura, R Yagi, Y Mazaki, E Schaefer, H Yano, H Sabe  JOURNAL OF CELL BIOLOGY  159-  (4)  673  -683  2002/11  [Not refereed][Not invited]
     
    RhoA activity is transiently inhibited at the initial phase of integrin engagement, when Cdc42- and/or Rac1-mediated membrane spreading and ruffling predominantly occur. Paxillin, an integrin-assembly protein, has four major tyrosine phosphorylation sites, and the phosphorylation of Tyr31 and Tyrl 18 correlates with cell adhesion and migration. We found that mutation of Tyr31/118 caused enhanced activation of RhoA and premature formation of stress fibers with substantial loss of efficient membrane spreading and ruffling in adhesion and migration of NMuMG cells. These phenotypes were similar to those induced by RhoA(G14V) in parental cells, and could be abolished by expression of RhoA(T19N), Rac1(G12V), or p190RhoGAP in the mutant-expressing cells. Phosphorylated Tyr31/118 was found to bind to two src homology (SH)2 domains of p120RasGAP, with coprecipitation of endogenous paxillin with p120RasGAP. p190RhoGAP is known to be a major intracellular binding partner for the p120RasGAP SH2 domains. We found that Tyr31/118-phosphorylated paxillin competes with p190RhoGAP for binding to p120RasGAP, and provides evidence that p190RhoGAP freed from p120RasGAP efficiently suppresses RhoA activity during cell adhesion. We conclude that Tyr31/118-phosphorylated paxillin serves as a template for the localized suppression of RhoA activity and is necessary for efficient membrane spreading and ruffling in adhesion and migration of NMuMG cells.
  • 低分子量G蛋白質Ralの下流分子であるPOB1は,パキシリン結合蛋白質PAG2と結合し,細胞運動を制御する
    大城 望史, 小山 眞也, 杉山 真一郎, 佐邊 壽孝, 菊池 章  生化学  74-  (8)  1049  -1049  2002/08  [Not refereed][Not invited]
  • 細胞外マトリックス系による細胞増殖と機能の制御 運動中の細胞における,パキシリンのチロシン31及び118のリン酸化により制御されるRhoA活性抑制の局在化機構
    坪内 朝子, 坂倉 純子, 八木 良平, 真崎 雄一, Schaefer Erik, 矢野 元, 佐邊 壽孝  日本発生生物学会大会講演要旨集  35回-  98  -98  2002/05  [Not refereed][Not invited]
  • 癌細胞の浸潤におけるパキシリンのチロシンリン酸化の役割
    岩崎 輝夫, 向井 睦子, 矢野 元, 佐邊 壽孝, 杉原 綾子, 山田 直子, 辻村 亨, 寺田 信行  日本病理学会会誌  91-  (1)  197  -197  2002/03  [Not refereed][Not invited]
  • Ensemble of membrane and cytoskeleton remodelings in cell migration
    Seikagaku  74(12):1429-40-  2002  [Not refereed][Not invited]
  • A. J. Woods, M. S. Roberts, J. Choudhary, S. T. Barry, Y. Mazaki, H. Sabe, S. J. Morley, D. R. Critchley & J. C. Norman. Paxillin associates with poly(A)-binding protein lat the dense ER and the leading edge of migrating cells
    J. Biol. Chem.  227-  6428  -6437  2002  [Not refereed][Not invited]
  • T Iwasaki, A Nakata, M Mukai, K Shinkai, H Yano, H Sabe, E Schaefer, M Tatsuta, T Tsujimura, N Terada, E Kakishita, H Akedo  INTERNATIONAL JOURNAL OF CANCER  97-  (3)  330  -335  2002/01  [Not refereed][Not invited]
     
    We demonstrated previously that rat ascites hepatoma MMI cells require both lysophosphatidic acid (LPA) and fibronectin (FN) for phagokinetic motility and transcellular migration and that these events are regulated through the RhoA-ROCK pathway. It remains to be elucidated, however, how the signals from both LPA and FN are integrated into cell migration. To examine this, total cellular lysates after stimulation with LPA or FN were subjected to time-course immunoblot analysis with anti-phosphotyrosine antibodies (Abs). Consequently, tyrosine-phosphorylation of paxillin was obviously persistent after stimulation with FN + LPA as compared to after stimulation with either alone. Tyrosine-phosphorylated paxillin comprised 2 components; slowly and fast migrating ones. Immunoblotting of anti-paxillin immunoprecipitates with phosphorylation site-specific Abs revealed the following: tyrosine-phosphorylation was enhanced preferentially on a slowly migrating component after stimulation with FN + LPA; this component contained phosphorylation at both tyrosine residue (Y) 31 and Y118; and phosphorylation of paxillin at Y181 was constitutive and not augmented by stimulation with either FN or LPA. Amiloride, an inhibitor of the Na+/H+ antiporter downstream of ROCK, suppressed cell motility and correspondingly paxillin tyrosine-phosphorylation at both Y31 and Y118. Paxillin phosphorylation weakly induced by FN alone, insufficient for cell migration, was not inhibited by amiloride. These results demonstrate that LPA collaborates with FN for persistent tyrosine phosphorylation of paxillin at both Y31 and Y118, regulated by the Na+/H+ antiporter downstream of ROCK and that this phosphorylated paxillin is essential for MMI cancer cell migration. (C) 2002 Wiley-Liss, Inc.
  • Ensemble of membrane and cytoskeleton remodelings in cell migration
    Seikagaku  74(12):1429-40-  2002  [Not refereed][Not invited]
  • A. J. Woods, M. S. Roberts, J. Choudhary, S. T. Barry, Y. Mazaki, H. Sabe, S. J. Morley, D. R. Critchley & J. C. Norman. Paxillin associates with poly(A)-binding protein lat the dense ER and the leading edge of migrating cells
    J. Biol. Chem.  227-  6428  -6437  2002  [Not refereed][Not invited]
  • T Iwasaki, A Nakata, M Mukai, K Shinkai, H Yano, H Sabe, E Schaefer, M Tatsuta, T Tsujimura, N Terada, E Kakishita, H Akedo  INTERNATIONAL JOURNAL OF CANCER  97-  (3)  330  -335  2002/01  [Not refereed][Not invited]
     
    We demonstrated previously that rat ascites hepatoma MMI cells require both lysophosphatidic acid (LPA) and fibronectin (FN) for phagokinetic motility and transcellular migration and that these events are regulated through the RhoA-ROCK pathway. It remains to be elucidated, however, how the signals from both LPA and FN are integrated into cell migration. To examine this, total cellular lysates after stimulation with LPA or FN were subjected to time-course immunoblot analysis with anti-phosphotyrosine antibodies (Abs). Consequently, tyrosine-phosphorylation of paxillin was obviously persistent after stimulation with FN + LPA as compared to after stimulation with either alone. Tyrosine-phosphorylated paxillin comprised 2 components; slowly and fast migrating ones. Immunoblotting of anti-paxillin immunoprecipitates with phosphorylation site-specific Abs revealed the following: tyrosine-phosphorylation was enhanced preferentially on a slowly migrating component after stimulation with FN + LPA; this component contained phosphorylation at both tyrosine residue (Y) 31 and Y118; and phosphorylation of paxillin at Y181 was constitutive and not augmented by stimulation with either FN or LPA. Amiloride, an inhibitor of the Na+/H+ antiporter downstream of ROCK, suppressed cell motility and correspondingly paxillin tyrosine-phosphorylation at both Y31 and Y118. Paxillin phosphorylation weakly induced by FN alone, insufficient for cell migration, was not inhibited by amiloride. These results demonstrate that LPA collaborates with FN for persistent tyrosine phosphorylation of paxillin at both Y31 and Y118, regulated by the Na+/H+ antiporter downstream of ROCK and that this phosphorylated paxillin is essential for MMI cancer cell migration. (C) 2002 Wiley-Liss, Inc.
  • Y. Mazaki, S. Hashimoto, K. Okawa, A. Tsubouchi, K. Nakamura, R. Yagi, H. Yano, A. Kondo, A. Iwamatsu, A. Mizoguchi, H. Sabe  Molecular Biology of the Cell  12-  645  -662  2001/11/22  [Not refereed][Not invited]
     
    Paxillin acts as an adaptor protein in integrin signaling. We have shown that paxillin exists in a relatively large cytoplasmic pool, including perinuclear areas, in addition to focal complexes formed at the cell periphery and focal adhesions formed underneath the cell. Several ADP-ribosylation factor (ARF) GTPase-activating proteins (GAPs; ARFGAPs) have been shown to associate with paxillin. We report here that Git2-short/KIAA0148 exhibits properties of a paxillin-associated ARFGAP and appears to be colocalized with paxillin, primarily at perinuclear areas. A fraction of Git2-short was also localized to actin-rich structures at the cell periphery. Unlike paxillin, however, Git2-short did not accumulate at focal adhesions underneath the cell. Git2-short is a short isoform of Git2, which is highly homologous to p95PKL, another paxillin-binding protein, and showed a weaker binding affinity toward paxillin than that of Git2. The ARFGAP activities of Git2 and Git2-short have been previously demonstrated in vitro, and we provided evidence that at least one ARF isoform, ARF1, is an intracellular substrate for the GAP activity of Git2-short. We also showed that Git2-short could antagonize several known ARF1-mediated phenotypes: overexpression of Git2-short, but not its GAP-inactive mutant, caused the redistribution of Golgi protein β-COP and reduced the amounts of paxillin-containing focal adhesions and actin stress fibers. Perinuclear localization of paxillin, which was sensitive to ARF inactivation, was also affected by Git2-short overexpression. On the other hand, paxillin localization to focal complexes at the cell periphery was unaffected or even augmented by Git2-short overexpression. Therefore, an ARFGAP protein weakly interacting with paxillin, Git2-short, exhibits pleiotropic functions involving the regulation of Golgi organization, actin cytoskeletal organization, and subcellular localization of paxillin, all of which need to be coordinately regulated during integrin-mediated cell adhesion and intracellular signaling.
  • Ralの下流分子であるPOB1とARF-GAPとの結合による細胞運動の制御
    大城 望史, 小山 眞也, 近藤 明子, 佐邊 壽孝, 菊池 章  生化学  73-  (8)  811  -811  2001/08  [Not refereed][Not invited]
  • 上皮細胞運動におけるPyk2とFakの活性制御とシグナル伝達の解析
    矢野 元, 中村 邦明, Schaefer Erik, 佐邊 壽孝  生化学  73-  (8)  811  -811  2001/08  [Not refereed][Not invited]
  • 細胞接着分子研究の進歩 細胞運動における細胞骨格再構成と形質膜再構成との統御機構
    佐邊 壽孝  炎症・再生  21-  (4)  443  -443  2001/07  [Not refereed][Not invited]
  • Kuniaki Nakamura, Hajime Yano, Erik Schaefer, Hisataka Sabe, Hisataka Sabe  Oncogene  20-  (21)  2626  -2635  2001/05/10  [Not refereed][Not invited]
     
    Integrin signaling is activated during epithelial-mesenchymal transdifferentiation (EMT) and cell migration, processes serving as models for carcinogenesis. We have shown that paxillin and p130Cas become highly tyrosine phosphorylated during these processes in NMuMG cells. Here, we examined the regulation of Fak and Pyk2, kinases implicated in this phosphorylation. Pyk2 became phosphorylated at the major autophosphorylation site (Tyr-402) and the potential Grb2-binding site (Tyr-881) during EMT. In contrast, phosphorylation of Fak at the corresponding autophosphorylation site (Tyr-397) occurred even in sedentary epithelial cells, whereas phosphorylation at Tyr-407 and Tyr-861 was induced during EMT. During cell migration, these phosphorylation events, except Fak Tyr-397, were augmented further, and phosphorylation of Fak Tyr-577 and the corresponding Pyk2 Tyr-580, both within the kinase activation loops, was also induced. In all cases, phosphorylation of the putative Grb2-binding site in Fak (Tyr-925) was almost undetectable. Although Fak and Pyk2 have several phosphorylation sites in common, Tyr-407 and Tyr-861 are unique to Fak. Our results revealed that Fak and Pyk2 are non-equivalent in the tyrosine phosphorylation events and thereby likely to evoke different downstream signaling cascades during EMT and cell migration of NMuMG cells. We also show that Fak Tyr-397 phosphorylation occurs exclusively at the cytoplasm, but not at focal contacts, in the sedentary epithelial cells. In contrast, all other tyrosine phosphorylated forms of Fak and Pyk2 are predominantly localized to focal adhesions and the cell periphery in motile cells, all colocalized with paxillin and p130Cas.
  • ダイナミックな細胞骨格制御と細胞機能 パキシリン結合性ARFGAP蛋白質のアクチン細胞骨格制御における役割
    真崎 雄一, 佐邊 壽孝  日本細胞生物学会大会講演要旨集  54回-  3  -3  2001/05  [Not refereed][Not invited]
  • マトリックス分子シグナルの多様性と細胞接着 細胞運動制御おけるチロシンリン酸化の役割
    矢野 元, 坪内 朝子, 佐邊 壽孝  日本細胞生物学会大会講演要旨集  54回-  37  -37  2001/05  [Not refereed][Not invited]
  • H Uchida, A Kondo, Y Yoshimura, Y Mazaki, H Sabe  JOURNAL OF EXPERIMENTAL MEDICINE  193-  (8)  955  -966  2001/04  [Not refereed][Not invited]
     
    The Fc gamma receptor (Fc gammaR)-mediated phagocytosis of macrophages is a complex process where remodeling of both the actin-based cytoskeleton and plasma membrane occur coordinately. Several different families of small GTPases are involved. We have isolated a GTPase-activating protein (GAP) for ADP-ribosylation factor (ARF), paxillin-associated protein with ARFGAP activity (PAG)3/Pap alpha /KIAA0400, from mature monocytes and macrophage-like cells. Mam malian ARFs fall into three classes, and the class III isoform (ARF6) has been shown to be involved in Fc gammaR-mediated phagocytosis. Here we report that PAG3 is enriched together with ARF6 and F-actin at phagocytic cups formed beneath immunoglobulin G-opsonized beads in P388D1 macrophages, in which overexpression of ARF6, but not ARF1 (class I) or ARF5 (class II), inhibits the phagocytosis. Overexpression of PAG3, but riot its GAP-inactive mutant, attenuated the focal accumulation of F-actin and blocked phagocytosis, although surface levels of the Fc gamma Rs were not affected. Other ubiquitously expressed ARFGAPs, G protein-coupled receptor kinase interactors GIT2 and GIT2-short/KIAA0148, which we have shown to exhibit GAP activity for ARF1 in COS-7 cells, did not accumulate at the phagocytic cups or inhibit phagocytosis. Moreover, cooverexpression of ARF6, but not ARF1 or ARF5, restored the phagocytic activity of PAG3-overexpressing cells. We propose that PAG3 acts as a GAP for ARF6 and is hence involved in Fc gammaR-mediated phagocytosis in mouse macrophages.
  • Shigeru Hashimoto, Asako Tsubouchi, Asako Tsubouchi, Yuichi Mazaki, Hisataka Sabe, Hisataka Sabe, Hisataka Sabe  Journal of Biological Chemistry  276-  (8)  6037  -6045  2001/02/23  [Not refereed][Not invited]
     
    p21-activated kinases (PAKs) are implicated in integrin signalings, and have been proposed to associate with paxillin indirectly. We show here that paxillin can bind directly to PAK3. We examined several representative focal adhesion proteins, and found that paxillin is the sole protein that associates with PAK3. PAK3 associated with the α and β isoforms of paxillin, but not with γ. We also show that paxillin α associated with both the kinase-inactive and the Cdc42-activated forms of PAK3 in vivo, without affecting the activation states of the kinase. A number of different functions have been ascribed to PAKs; and PAKs can bind directly to growth factor signaling-adaptor molecule, Nck, anda and a guanine nucleotide exchanger, βPIX. Our results revealed that paxillin α can compete with Nck and βPIX in the binding of PAK3. Moreover, paxillin α can be phosphorylated by PAK3 at serine. Therefore, paxillin α but not γ, appears to be capable of linking both the kinase-inactive and activated forms of PAK3 to integrins independent of Nck and βPIX, as Nck links PAK1 to growth factor receptors. Our results also revealed that paxillin is involved in highly complexed protein-protein interactions in integrin signaling.
  • EMBO report  2-  (814-820)  2001  [Not refereed][Not invited]
  • An ADP-Ribosylation Factor GTPase-activating Protein Git2-short/KIAA0148 Is Involved in Subcellular Localization of Paxillin and Actin Cyeoskeletal Organization.(共著)
    Mol.Biol.Cell  12-  645  -662  2001  [Not refereed][Not invited]
  • EMBO report  2-  (814-820)  2001  [Not refereed][Not invited]
  • An ADP-Ribosylation Factor GTPase-activating Protein Git2-short/KIAA0148 Is Involved in Subcellular Localization of Paxillin and Actin Cyeoskeletal Organization.(共著)
    Mol.Biol.Cell  12-  645  -662  2001  [Not refereed][Not invited]
  • A Suzuki, N Kadota, T Hara, Y Nakagami, T Izumi, T Takenawa, H Sabe, T Endo  ONCOGENE  19-  (51)  5842  -5850  2000/11  [Not refereed][Not invited]
     
    Meltrin alpha /ADAM12 is a member of the ADAM/MDC family proteins characterized by the presence of metalloprotease and disintegrin domains. This protein also contains a single transmembrane domain and a relatively long cytoplasmic domain containing several proline-rich sequences. These sequences are compatible with the consensus sequences for binding the Src homology 3 (SH3) domains. To determine whether the proline-rich sequences interact with SH3 domains in several proteins, binding of recombinant SH3 domains to the meltrin alpha cytoplasmic domain was analysed by pull-down assays. The SH3 domains of Src and Yes bound strongly, but that of Abl or phosphatidylinositol 3-kinase p85 subunit did not. Full-length Grb2/Ash bound strongly, whereas its N-terminal SH3 domain alone did less strongly. Src and Grb2 in bovine brain extracts also bound to meltrin alpha cytoplasmic domain on affinity resin, Furthermore, immunoprecipitation with alpha monoclonal antibody to meltrin alpha resulted in coprecipitation of Src and Grb2 with meltrin alpha in cell extracts, suggesting that Src and Grb2 are associated in vivo with meltrin alpha cytoplasmic domain. This notion was also supported by the findings that exogenously expressed meltrin cytoplasmic domain coexisted with Src and Grb2 on the membrane ruffles. The C-terminal Tyr901 of meltrin alpha was phosphorylated both initio and in cultured cells by v-Src. These results may imply that meltrin alpha cytoplasmic domain is involved in a signal transduction for some biological function through the interaction with SH3-containing proteins.
  • K Nakamura, H Yano, H Uchida, S Hashimoto, E Schaefer, H Sabe  JOURNAL OF BIOLOGICAL CHEMISTRY  275-  (35)  27155  -27164  2000/09  [Not refereed][Not invited]
     
    Temporal and spatial regulation of actin-based cytoskeletal organization and focal adhesion formation play an essential role in cell migration. Here, we show that tyrosine phosphorylation of a focal adhesion protein, paxillin, crucially participates in these regulations. We found that tyrosine phosphorylation of paxillin was a prominent event upon integrin activation during epithelial-mesenchymal trans differentiation and cell migration. Four major tyrosine phosphorylation sites were identified, and two of them were highly inducible upon integrin activation. Paxillin exhibits three distinct subcellular localizations as follows: localization along the cell periphery colocalized with circumferential actin meshworks, macroaggregation at focal adhesions connected to actin stress fibers, and diffuse cytoplasmic distribution. Tyrosine phosphorylation of paxillin localized at the cell periphery and focal adhesions was shown using phosphorylation site-specific antibodies. Mutations in the phosphorylation sites affected the peripheral localization of paxlllin and paxillin-containing focal adhesion formation during cell migration and cell-cell collision, accompanied by altered actin organizations. Our analysis indicates that phosphorylation of multiple tyrosines in paxillin cu is necessary for the proper function of paxillin and is involved in the temporospatial regulation of focal adhesion formation and actin cytoskeletal organization in motile cells.
  • パキシリンとp130Casは,そのチロシンリン酸化を介し,細胞運動調節とcontact inhibition of growthにおいて相反的に作用する
    矢野 元, 内田 浩, 岩崎 輝夫, 向井 睦子, 明渡 均, 中村 邦明, 佐邊 壽孝  日本癌学会総会記事  59回-  135  -135  2000/09  [Not refereed][Not invited]
  • K Nakamura, H Yano, H Uchida, S Hashimoto, E Schaefer, H Sabe  JOURNAL OF BIOLOGICAL CHEMISTRY  275-  (35)  27155  -27164  2000/09  [Not refereed][Not invited]
     
    Temporal and spatial regulation of actin-based cytoskeletal organization and focal adhesion formation play an essential role in cell migration. Here, we show that tyrosine phosphorylation of a focal adhesion protein, paxillin, crucially participates in these regulations. We found that tyrosine phosphorylation of paxillin was a prominent event upon integrin activation during epithelial-mesenchymal trans differentiation and cell migration. Four major tyrosine phosphorylation sites were identified, and two of them were highly inducible upon integrin activation. Paxillin exhibits three distinct subcellular localizations as follows: localization along the cell periphery colocalized with circumferential actin meshworks, macroaggregation at focal adhesions connected to actin stress fibers, and diffuse cytoplasmic distribution. Tyrosine phosphorylation of paxillin localized at the cell periphery and focal adhesions was shown using phosphorylation site-specific antibodies. Mutations in the phosphorylation sites affected the peripheral localization of paxlllin and paxillin-containing focal adhesion formation during cell migration and cell-cell collision, accompanied by altered actin organizations. Our analysis indicates that phosphorylation of multiple tyrosines in paxillin cu is necessary for the proper function of paxillin and is involved in the temporospatial regulation of focal adhesion formation and actin cytoskeletal organization in motile cells.
  • H Yano, H Uchida, T Iwasaki, M Mukai, H Akedo, K Nakamura, S Hashimoto, H Sabe  PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA  97-  (16)  9076  -9081  2000/08  [Not refereed][Not invited]
     
    Protein tyrosine phosphorylation accompanies and is essential for integrin signaling. We have shown that tyrosine phosphorylation of paxillin alpha and Crk-associated substrate (p130(Cas)) is a prominent event on integrin activation in normal murine mammary gland epithelial cells. Tyrosine phosphorylation of p130(Cas) has been demonstrated to facilitate cell migration. We show here that tyrosine phosphorylation of paxillin alpha acts to reduce haptotactic cell migrations as well as transcellular invasive activities in several different experimental cell systems, whereas tyrosine phosphorylation of p130(Cas) exerts opposing effects to those of paxillin alpha, Each of the phosphorylation-null mutants acts as a dominant negative for each phenotype, Moreover, we found that overexpression of paxillin alpha reduced the cell saturation density of normal murine mammary gland cells, whereas overexpression of p130(Cas) increased it. These effects also seemed to depend on tyrosine phosphorylation events, Cell growth rates and morphologies at growing phases were not significantly altered, nor were cells transformed. Addition of epidermal growth factor increased saturation density of the paxillin alpha-overexpressing cells, whereas no further increment was observed in p130(Cas)-overexpressing cells. We propose that tyrosine phosphorylation of paxillin alpha and p130(Cas) exerts opposing effects on several integrin-mediated cellular events, possibly through different signaling pathways.
  • スフィンゴ脂質蓄積症における脳内巨大多核細胞産生機構の解析
    金沢 崇之, 中村 祥子, 桃井 道子, 山地 俊之, 竹松 弘, 矢野 元, 佐邊 壽孝, 山本 章嗣, 川嵜 敏祐, 小堤 保則  生化学  72-  (8)  713  -713  2000/08  [Not refereed][Not invited]
  • 細胞接着と細胞骨格の制御と細胞形態形成 細胞骨格制御におけるパキシリンと低分子量G蛋白質群との機能連関
    佐邊 壽孝, 橋本 茂, 近藤 明子, 坪内 朝子, 内田 浩, 中村 邦明, 矢野 元, 真崎 雄一  生化学  72-  (8)  593  -593  2000/08  [Not refereed][Not invited]
  • T Kanazawa, S Nakamura, M Momoi, T Yamaji, H Takematsu, H Yano, H Sabe, A Yamamoto, T Kawasaki, Y Kozutsumi  JOURNAL OF CELL BIOLOGY  149-  (4)  943  -950  2000/05  [Not refereed][Not invited]
     
    Although a number of cellular components of cytokinesis have been identified, little is known about the derailed mechanisms underlying this process. Here, we report that the lipid metabolite psychosine (galactosylsphingosine), derived from galactosylceramide, induced formation of multinuclear cells from a variety of nonadherent and adherent cells due to inhibition of cytokinesis, When psychosine was added to the human myelomonocyte cell line U937, which was the most sensitive among the cell lines tested, cleavage furrow formed either incompletely or almost completely. However, abnormal contractile movement was detected in which the cellular contents of one of the hemispheres of the contracting cell were transferred into its counterpart. Finally, the cleavage furrow disappeared and cytokinesis was reversed. Psychosine treatment also induced giant clots of actin filaments in the cells that probably consisted of small vacuoles with filamentous structures, suggesting that psychosine affected actin reorganization, These observations could account for the formation of multinuclear globoid cells in the brains of patients with globoid eel leukodystrophy, a neurological disorder characterized by the accumulation of psychosine due to galactosylceramidase deficiency.
  • A Kondo, S Hashimoto, H Yano, K Nagayama, Y Mazaki, H Sabe  MOLECULAR BIOLOGY OF THE CELL  11-  (4)  1315  -1327  2000/04  [Not refereed][Not invited]
     
    Paxillin acts as an adaptor molecule in integrin signaling. Paxillin is localized to focal contacts but seems to also exist in a relatively large cytoplasmic pool. Here, we report the identification of a new paxillin-binding protein, FAGS (paxillin-associated protein with ADP-ribosylation factor [ARF] GTPase-activating protein [GAP] activity, number 3), which is involved in regulation of the subcellular localization of paxillin. FAGS bound to all paxillin isoforms and was induced during monocyte maturation, at which time paxillin expression is also increased and integrins are activated. PAG3 was diffusely distributed in the cytoplasm in premature monocytes but became localized at cell periphery in mature monocytes, a fraction of which then colocalized with paxillin. PAG3, on the other hand, did not accumulate at focal adhesion plaques, suggesting that PAG3 is not an integrin assembly protein. PAG3 was identical to KIAA0400/Pap alpha, which was previously identified as a Pyk2-binding protein bearing a GAP activity toward several ARFs in vitro. Mammalian ARFs fall into three classes, and we showed that all classes could affect subcellular localization of paxillin. We also examined possible interaction of FAGS with ARFs and showed evidence that at least one of them, ARF6, seems to be an intracellular substrate for GAP activity of FAGS. Moreover, overexpression of PAG3, but not its GAP-inactive mutant, inhibited paxillin recruitment to focal contacts and hampered cell migratory activities, whereas cell adhesion activities were almost unaffected. Therefore, our results demonstrate that paxillin recruitment to focal adhesions is not mediated by simple cytoplasmic diffusion; rather, PAG3 appears to be involved in this process, possibly through its GAP activity toward ARF proteins. Our result thus delineates a new aspect of regulation of cell migratory activities.
  • ヒト子宮内膜間質細胞脱落膜化過程における接着斑分子の動態
    丸山 哲夫, 吉村 泰典, 野澤 志朗, 佐邊 壽孝  日本産科婦人科学会雑誌  52-  (2)  334  -334  2000/02  [Not refereed][Not invited]
  • 丸山 哲夫, 吉村 〓典, 野澤 志朗, 佐邊 壽孝  日本産科婦人科學會雜誌  52-  (2)  "334(S  -258)"  2000/02/01  [Not refereed][Not invited]
  • KANAI F, MARIGNANI P A, SARBASSOVA D, YAGI R, HALL R A, DONOWITZ M, HISAMINATO A, FUJIWARA T, YAFFE M B  EMBO J.  19-  (24)  6778  -6791  2000  [Not refereed][Not invited]
  • A new paxillin-binding protein, PAG3/Pap αKIAA0400, bearing an ARF GTPase-activating protein activity, is involved in paxillin recruitment to focal adhesions and cell migration.(共著)
    Mol.Biol.Cell  11-  1315  -1327  2000  [Not refereed][Not invited]
  • A new paxillin-binding protein, PAG3/Papd/KIAA 0400, bearing an Arf GTPase-activating protein activity is involved in paxillin recruitment to focal adhesions and cell migration.
    Mol. Biol. Cell  11-  1315  -1327  2000  [Not refereed][Not invited]
  • ITO A, KATAOKA T R, WATANABE M, NISHIYAMA K, MAZAKI Y, SABE H, KITAMURA Y, NOJIMA H  EMBO J.  19-  (4)  562  -571  2000  [Not refereed][Not invited]
  • A new paxillin-binding protein, PAG3/Pap αKIAA0400, bearing an ARF GTPase-activating protein activity, is involved in paxillin recruitment to focal adhesions and cell migration.(共著)
    Mol.Biol.Cell  11-  1315  -1327  2000  [Not refereed][Not invited]
  • A new paxillin-binding protein, PAG3/Papd/KIAA 0400, bearing an Arf GTPase-activating protein activity is involved in paxillin recruitment to focal adhesions and cell migration.
    Mol. Biol. Cell  11-  1315  -1327  2000  [Not refereed][Not invited]
  • EMBO J.  19-  (4)  562  -571  2000  [Not refereed][Not invited]
  • Tetsuo Maruyama, Tetsuo Maruyama, Tetsuo Maruyama, Yasunori Yoshimura, Junji Yodoi, Hisataka Sabe, Hisataka Sabe  Endocrinology  140-  2632  -2636  1999/12/01  [Not refereed][Not invited]
     
    Tyrosine phosphorylation of cellular proteins, controlled coordinately by tyrosine kinases and phosphatases, is a critical element in signal transduction pathways involved in the regulation of biological responses including cell growth and differentiation. Decidualization is a dramatic progesterone-induced differentiation of the estrogen-primed endometrium, which is crucial for embryo implantation and maintenance of pregnancy. Here we have shown that the kinase activity of c-Src was increased, accompanied by altered tyrosine phosphorylation of several cellular proteins, during in vitro decidualization of human endometrial stromal cells. Withdrawal of both estrogen and progesterone from the cultures of decidualized stromal cells reduced c-Src kinase activity to the basal level and also changed the pattern of tyrosine phosphorylation of the several cellular proteins to the unstimulated state. The kinase activity of endometrial c-Src appeared to inversely correlate with the level of its tyrosine phosphorylation. Moreover, although the endometrial stromal cells expressed another src-family kinase, Fyn, the activity of the Fyn kinase was almost undetectable during decidualization and thereafter upon steroid withdrawal. Our findings suggest that the activation of c-Src kinase may be a normal physiological event associated with decidualization, being specifically involved in the signaling cascades mediated by ovarian hormone stimulation.
  • T Maruyama, Y Yoshimura, H Sabe  ENDOCRINOLOGY  140-  (12)  5982  -5990  1999/12  [Not refereed][Not invited]
     
    Human endometrial stromal cells undergo in vitro decidualization when treated with progesterone and estrogen. Using this model, we previously reported specific changes in the c-Src kinase activity and tyrosine phosphorylation of several proteins during in vitro decidualization, Focal adhesion kinase (FAK) and paxillin are known to form a complex with c-Src at the focal contacts and to participate in the integrin-mediated signal transduction as c-Src substrates. We here examined the tyrosine phosphorylation and subcellular localization of the focal adhesion proteins in stromal cells isolated from human endometrium. We found, however, that the total levels of FAK and paxillin tyrosine phosphorylation were not markedly changed during decidualization or after steroid withdrawal. In our culture system, numerous multicellular nodules were developed in cultures of decidualized stromal cells, within whose nodules the focal contacts were found to disappear. Moreover, disruption of the focal contacts was accompanied by disorganization of the actin-based cytoskeleton. These findings suggest that tyrosine phosphorylation of the endometrial paxillin and FBR is not tightly regulated by the kinase activity of c-Src during in vitro decidualization. The escape from regulation by c-Src may be in part due to the dissociation of the focal adhesion proteins/c-Src complex caused by the breakdown of the focal adhesion plaques as well as the loss of the actin-based cytoskeletal architecture.
  • T Hiiragi, H Sasaki, A Nagafuchi, H Sabe, SC Shen, M Matsuki, K Yamanishi, S Tsukita  JOURNAL OF BIOLOGICAL CHEMISTRY  274-  (48)  34148  -34154  1999/11  [Not refereed][Not invited]
     
    Transglutaminase type 1 was identified as a tyrosinephosphorylated protein from the isolated junctional fraction of the mouse liver. This enzyme was reported to be involved in the covalent cross-linking of proteins in keratinocytes, but its expression and activity in other cell types have not been examined. Northern blotting revealed that transglutaminase type 1 was expressed in large amounts in epithelial tissues (lung, liver, and kidney), which was also confirmed by immunoblotting with antibodies raised against mouse recombinant protein. Immunoblotting of the isolated junctional fraction revealed that transglutaminase type 1 was concentrated in the fraction not only as a 97-kDa form but also as forms of various molecular masses cross-linked to other proteins. In agreement with this finding, endogenous transglutaminase type 1 was immunofluorescently colocalized with E-cadherin in cultured simple epithelial cells. In the liver and kidney, immunoelectron microscopy revealed that transglutaminase type 1 was concentrated, albeit not exclusively, at cadherin-based adherens junctions. Furthermore, by in vitro and in vivo labeling, transglutaminase cross-linking activity was also shown to be concentrated at intercellular junctions of simple epithelial cells. These findings suggested that the formation of covalently cross-linked multimolecular complexes by transglutaminase type 1 is an important mechanism for maintenance of the structural integrity of simple epithelial cells, especially at cadherin-based adherens junctions.
  • MCP-1受容体を介した情報伝達におけるチロシンキナーゼPyk2の役割の検討
    山崎 雅秀, 荒井 秀典, 石井 賢二, 北 徹, 佐邊 壽孝  動脈硬化  27-  (Suppl.1)  145  -145  1999/11  [Not refereed][Not invited]
  • 細胞斉一単層形成能とパキシリンの機能
    佐邊 壽孝  日本癌学会総会記事  58回-  39  -39  1999/08  [Not refereed][Not invited]
  • ARF GAP活性を有するPagはゴルジ構造とパキシリンの細胞内局在制御に関与する
    真崎 雄一, 矢野 元, 大川 克也, 岩松 明彦, 佐邊 壽孝  日本癌学会総会記事  58回-  184  -184  1999/08  [Not refereed][Not invited]
  • ARF GAP活性を有するパキシリン結合性新規タンパク質
    近藤 明子, 橋本 茂, 真崎 雄一, 佐邊 壽孝  日本癌学会総会記事  58回-  184  -184  1999/08  [Not refereed][Not invited]
  • 【シグナル伝達の場としての細胞膜ドメイン】 細胞接着形成と細胞内小胞輸送活性
    近藤 明子, 佐邊 壽孝  細胞工学  18-  (8)  1141  -1147  1999/08  [Not refereed][Not invited]
  • パキシリンを介する細胞運動の接触阻止機構の解析
    中村 邦明, 矢野 元, 佐邊 壽孝  日本癌学会総会記事  58回-  234  -234  1999/08  [Not refereed][Not invited]
  • パキシリンのチロシンリン酸化を介する細胞-細胞間接触検知機構の解析
    矢野 元, 中村 邦明, 内田 浩, 橋本 茂, 佐邊 壽孝  日本癌学会総会記事  58回-  234  -234  1999/08  [Not refereed][Not invited]
  • 細胞運動におけるパキシリンとp130casの相反的影響
    内田 浩, 明渡 均, 岩崎 輝夫, 佐邊 壽孝  日本癌学会総会記事  58回-  477  -477  1999/08  [Not refereed][Not invited]
  • ボディープランを司るチロシンリン酸化の意義 細胞運動における細胞間接触感知機構
    佐邊 壽孝  生化学  71-  (8)  656  -656  1999/08  [Not refereed][Not invited]
  • 近藤 明子, 佐邊 壽孝  細胞工学  18-  (8)  1141  -1147  1999/08  [Not refereed][Not invited]
  • T Maruyama, Y Yoshimura, J Yodoi, H Sabe  ENDOCRINOLOGY  140-  (6)  2632  -2636  1999/06  [Not refereed][Not invited]
     
    Tyrosine phosphorylation of cellular proteins, controlled coordinately by tyrosine kinases and phosphatases, is a critical element in signal transduction pathways involved in the regulation of biological responses including cell growth and differentiation. Decidualization is a dramatic progesterone-induced differentiation of the estrogen-primed endometrium, which is crucial for embryo implantation and maintenance of pregnancy. Here we have shown that the kinase activity of c-Src was increased, accompanied by altered tyrosine phosphorylation of several cellular proteins, during in vitro decidualization of human endometrial stromal cells. Withdrawal of both estrogen and progesterone from the cultures of decidualized stromal cells reduced c-Src kinase activity to the basal level and also changed the pattern of tyrosine phosphorylation of the several cellular proteins to the unstimulated stale. The kinase activity of endometrial c-Src appeared to inversely correlate with the level of its tyrosine phosphorylation. Moreover, although the endometrial stromal cells expressed another src-family kinase, Fyn, the activity of the Fyn kinase was almost undetectable during decidualization and thereafter upon steroid withdrawal. Our findings suggest that the activation of c-Src kinase may be a normal physiological event associated with decidualization, being specifically involved in the signaling cascades mediated by ovarian hormone stimulation.
  • Vascular endothelial growth factor induces activation and subcellular translocation of focal adhesion kinase (p125(FAK)) in cultured rat cardiac myocytes
    N Takahashi, Y Seko, E Noiri, K Tobe, T Kadowaki, H Sabe, Y Yazaki  CIRCULATION RESEARCH  84-  (10)  1194  -1202  1999/05  [Not refereed][Not invited]
     
    Vascular endothelial growth factor (VEGF) has been proposed to be among the candidate factors with the most potential to play a role in ischemia-induced collateral vessel formation, Recently, we found that VEGF activated the mitogen-activated protein kinase cascade in cultured rat cardiac myocytes, To elucidate how VEGF affects adhesive interaction of cardiac myocytes with the extracellular matrix (ECM), one of the important cell functions, we investigated the molecular mechanism of activation of focal adhesion-related proteins, especially focal adhesion kinase (p125(FAK)), in cultured rat cardiac myocytes, We found that the 2 VEGF receptors, KDR/Flk-1 and Flt-1, were expressed in cardiac myocytes and that KDR/Flk-1 was significantly tyrosine phosphorylated on VEGF stimulation, VEGF induced tyrosine phosphorylation and activation of p125(FAK) as well as tyrosine phosphorylation of paxillin; this was accompanied by subcellular translocation of p125(FAK) from perinuclear sites to the focal adhesions. This VEGF-induced activation of p125(FAK) was inhibited partially by the tyrosine kinase inhibitors genistein and tyrphostin, Activation of p125(FAK) was accompanied by its increased association with adapter proteins GRB2, Shc, and nonreceptor type tyrosine kinase p60(c-src) Furthermore, we confirmed that VEGF induced a significant increase in adhesive interaction between cardiac myocytes and ECM using an electric cell-substrate impedance sensor. These results strongly suggest that p125(FAK) is one of the most important components in VEGF-induced signaling in cardiac myocytes, playing a critical role in adhesive interaction between cardiac myocytes and ECM.
  • Vascular endothelial growth factor induces activation and subcellular translocation of focal adhesion kinase (p125(FAK)) in cultured rat cardiac myocytes
    N Takahashi, Y Seko, E Noiri, K Tobe, T Kadowaki, H Sabe, Y Yazaki  CIRCULATION RESEARCH  84-  (10)  1194  -1202  1999/05  [Not refereed][Not invited]
     
    Vascular endothelial growth factor (VEGF) has been proposed to be among the candidate factors with the most potential to play a role in ischemia-induced collateral vessel formation, Recently, we found that VEGF activated the mitogen-activated protein kinase cascade in cultured rat cardiac myocytes, To elucidate how VEGF affects adhesive interaction of cardiac myocytes with the extracellular matrix (ECM), one of the important cell functions, we investigated the molecular mechanism of activation of focal adhesion-related proteins, especially focal adhesion kinase (p125(FAK)), in cultured rat cardiac myocytes, We found that the 2 VEGF receptors, KDR/Flk-1 and Flt-1, were expressed in cardiac myocytes and that KDR/Flk-1 was significantly tyrosine phosphorylated on VEGF stimulation, VEGF induced tyrosine phosphorylation and activation of p125(FAK) as well as tyrosine phosphorylation of paxillin; this was accompanied by subcellular translocation of p125(FAK) from perinuclear sites to the focal adhesions. This VEGF-induced activation of p125(FAK) was inhibited partially by the tyrosine kinase inhibitors genistein and tyrphostin, Activation of p125(FAK) was accompanied by its increased association with adapter proteins GRB2, Shc, and nonreceptor type tyrosine kinase p60(c-src) Furthermore, we confirmed that VEGF induced a significant increase in adhesive interaction between cardiac myocytes and ECM using an electric cell-substrate impedance sensor. These results strongly suggest that p125(FAK) is one of the most important components in VEGF-induced signaling in cardiac myocytes, playing a critical role in adhesive interaction between cardiac myocytes and ECM.
  • SEKO Y, TAKAHASHI N, SABE H, TOBE K, KADOWAKI T, NAGAI R  Biopby, Biochem.Res.Com  262-  (1)  290  -296  1999  [Not refereed][Not invited]
  • Tyrosine phosphorylation and subcelluar localization of focal adhesion proteins during in vitro decidualization of human endometrial stromal cells.
    Endocrinology.  140-  5982  -5990  1999  [Not refereed][Not invited]
  • SEKO Y, TAKAHASHI N, SABE H, TOBE K, KADOWAKI T, NAGAI R  Biophy. Biochem. Res. Com.  262-  (1)  290  -296  1999  [Not refereed][Not invited]
  • 細胞はどのようにして動くか
    JST基礎研究報 「源流」  (1)  13  -20  1999  [Not refereed][Not invited]
  • Biopby, Biochem.Res.Com  262-  (1)  290  -296  1999  [Not refereed][Not invited]
  • Tyrosine phosphorylation and subcelluar localization of focal adhesion proteins during in vitro decidualization of human endometrial stromal cells.
    Endocrinology.  140-  5982  -5990  1999  [Not refereed][Not invited]
  • Biophy. Biochem. Res. Com.  262-  (1)  290  -296  1999  [Not refereed][Not invited]
  • Endocrinology  140-  2632  -2636  1999  [Not refereed][Not invited]
  • 佐邊 壽孝  源流  1999-  (1)  13  -20  1999  [Not refereed][Not invited]
     
    上皮間充織細胞においては、その形質転換に伴ってカドヘリンの発現低下とインテグリンの発現上昇が観察される。本研究ではまず、当該細胞の形質転換における細胞接着性変化の分子機構を明らかにするために、カドヘリンとインテグリンの裏打ちタンパク質群について検討した。その結果、インテグリンの活性化に伴って2種のタンパク質でのチロシンリン酸化の亢進が観察され、最も顕著な変化を呈したタンパク質としてパキシリンが同定された。パキシリンのリン酸化部位の変異体cDNAを細胞に発現した際の効果を検討した結果、パキシリンのチロシンリン酸化が、この形質転換に伴う細胞運動性変化をもたらす主要因子であること、その複数部位のチロシンリン酸化が協調することにより細胞同士の「接触による増殖阻止」と「接触による運動性阻止」とを制御することを明らかにした。さらに、上皮細胞においては、パキシリンの複数部位のチロシンリン酸化による制御を乱すことにより、細胞同士が十分接触していてもカドヘリンを介した細胞間接着が正常には形成されないことも明らかにした。
  • UCHIDA Hiroshi, YANO Hajime, MAZAKI Yuichi, HASHIMOTO Shigeru, SABE Hisataka  日本分子生物学会年会プログラム・講演要旨集  21-  (0)  1998/12/01  [Not refereed][Not invited]
  • YANO Hajime, UCHIDA Hiroshi, SABE Hisataka  日本分子生物学会年会プログラム・講演要旨集  21-  (0)  1998/12/01  [Not refereed][Not invited]
  • MAZAKI Yuichi, YANO Hajime, OKAWA Katsuya, HASHIMOTO Shigeru, IWAMATSU Akihiro, SABE Hisatake  日本分子生物学会年会プログラム・講演要旨集  21-  (0)  1998/12/01  [Not refereed][Not invited]
  • KONDO Akiko, HASHIMOTO Shigeru, MAZAKI Yuichi, NAGAYAMA Kuniaki, SABE Hisataka  日本分子生物学会年会プログラム・講演要旨集  21-  (0)  1998/12/01  [Not refereed][Not invited]
  • HASHIMOTO Shigeru, MAZAKI Yuichi, UCHIDA Hirosi, SABE Hisataka  日本分子生物学会年会プログラム・講演要旨集  21-  (0)  1998/12/01  [Not refereed][Not invited]
  • N Nishiya, H Sabe, K Nose, M Shibanuma  NUCLEIC ACIDS RESEARCH  26-  (18)  4267  -4273  1998/09  [Not refereed][Not invited]
     
    hic-5 protein is a member of the LIM protein family, containing four LIM domains in its C-terminal region. It is mainly localized in focal adhesions and shows striking similarity to paxillin in its LIM domains, although the function of these LIM domains has remained elusive. In the present study, we found that full-length and the C-terminal half of hic-5 protein, including four LIM domains, bound to DNA in a zinc-dependent manner in vitro. Mouse genomic fragments that specifically bound to the hic-5-protein were isolated by successive rounds of hic-5 protein-DNA complex immunoprecipitation and PCR amplification. Seven independent clones were isolated, which contained high amounts of G+A and/or a long A/T tract. A DNA binding protein blot assay revealed the specificity of the interaction between hic-5 protein and the DNA fragment. Using a series of truncated forms of the hic-5 LIM domains, each of the four LIM domains was found to contribute to DNA binding in a distinctive manner.
  • Y Mazaki, H Uchida, O Hino, S Hashimoto, H Sabe  JOURNAL OF BIOLOGICAL CHEMISTRY  273-  (35)  22435  -22441  1998/08  [Not refereed][Not invited]
     
    Paxillin, a focal adhesion protein, exists as multiple isoforms in humans (alpha, beta, and gamma), To understand more about the physiological role of each isoform, we have employed the mouse system. We found that although the alpha and beta isoforms are present in the mouse, the gamma isoform is not. The alpha isoform protein was detected clearly in most adult tissues, whereas the beta isoform protein was almost undetectable except in spleen, testis, thymus, and lung. On the other hand, mRNAs of both isoforms were detectable in all tissues we examined. High levels of the beta isoform protein was detected in peritoneal exudate macrophage cells in adult mouse as well as in cultured fibroblasts, together with the alpha isoform, The alpha isoform was expressed at a constant level throughout the embryonic stages we examined, whereas the beta isoform protein was detected at the mid-stages of development and increased to levels almost equal to those of the a isoform during the late stages of embryogenesis, Therefore, unlike the alpha isoform, expression of the beta isoform protein is restricted in adult tissues. Moreover, we showed that alpha and beta isoforms were colocalized within the same focal adhesion plaques, and cytoplasmic pools of both isoforms exist in the perinuclear area, colocalized with the Gels apparatus.
  • 膜裏打ち分子による細胞内情報伝達 上皮間充織形質転換と細胞生存性・運動性の制御 インテグリン裏打ち蛋白質パキシリン(Paxillin)を中心として
    佐邊 壽孝, 真崎 雄一, 内田 浩, 橋本 茂  生化学  70-  (8)  703  -703  1998/08  [Not refereed][Not invited]
  • 接着斑タンパク質パキシリン(Paxillin)のゴルジ装置への局在について
    真崎 雄一, 大川 克也, 内田 浩, 橋本 茂, 岩松 明彦, 佐邊 壽孝  日本癌学会総会記事  57回-  146  -146  1998/08  [Not refereed][Not invited]
  • ヒト癌におけるpaxillin isoformの発現の解析
    内田 浩, 真崎 雄一, 橋本 茂, 佐邊 壽孝  日本癌学会総会記事  57回-  451  -451  1998/08  [Not refereed][Not invited]
  • 上皮系細胞のインテグリンを介する生存性維持機構に基付いた細胞癌化機構の解析
    橋本 茂, 真崎 雄一, 内田 浩, 佐邊 壽孝  日本癌学会総会記事  57回-  450  -450  1998/08  [Not refereed][Not invited]
  • Ryuji Yamaguchi, Ryuji Yamaguchi, Yuichi Mazaki, Kiichi Hirota, Shigeru Hashimoto, Hisataka Sabe, Hisataka Sabe  Oncogene  15-  (15)  1753  -1761  1997/11/19  [Not refereed][Not invited]
     
    Mitotic cells typically lack well-formed focal adhesions. As an approach to explore the dynamic process regulating the focal adhesion assembly, we examined states of focal adhesion proteins during mitosis of the cell cycle. We found that the amount of paxillin was significantly reduced during mitosis of the cell cycle, whereas other focal adhesion proteins including talin, vinculin and Focal Adhesion Kinase did not. Proteolytic degradation appeared to be involved in the mitotic reduction, but transcriptional and/or translational controls of the mRNA were not essential for this downregulation. Moreover, concurrent with the decreased protein level, phosphorylation status of paxillin altered during mitosis; mitotic paxillin was phosphorylated primarily on serine and dephosphorylated on tyrosine while interphase one was phosphorylated both on serine and tyrosine. We found that mitotic phosphorylation created an electrophoretically slow-migrating population of paxillin which was barely detected in interphase cells. This mitotic specific modification occurred with both α and β isoforms of paxillin. We also examined the fate of paxillin protein by changing its protein amount. We found that majority of paxillin overexpressed was subjected to the specific modification but not to the downregulation in the mitotic arrested cells. On the other hand, paxillin exogenously expressed at a moderate level was subjected to both the mitotic modification and downregulation. Collectively, we concluded that paxillin's specific serine phosphorylation together with the proteolytic downregulation of a limited fraction of paxillin is taken place during the mitosis of the cell cycle.
  • Sabe H  Biophysics  37-  (2)  1997/09/05  [Not refereed][Not invited]
  • 【サブメンブレンコート構造と膜蛋白質集積の機構】 インテグリン接着シグナル制御機構の多機能性・多様性創製の基本因子の解明にむけて
    佐邊 壽孝  生物物理  37-  (Suppl.2)  S9  -S9  1997/09  [Not refereed][Not invited]
  • がん遺伝子とシグナル伝達 細胞接着とシグナル伝達
    佐邊 壽孝  蛋白質・核酸・酵素  42-  (10)  1541  -1550  1997/07  [Not refereed][Not invited]
  • インテグリンを介した細胞基質間接着とシグナル伝達
    佐邊 壽孝  日本医師会雑誌  118-  (2)  225  -225  1997/07  [Not refereed][Not invited]
  • 佐辺 寿孝  蛋白質核酸酵素  42-  (10)  1541  -1550  1997/07  [Not refereed][Not invited]
  • Hisataka Sabe, Hisataka Sabe, Hisataka Sabe, Hisataka Sabe, Michinari Hamaguchi, Hidesaburo Hanafusa  Oncogene  14-  (15)  1779  -1788  1997/05/28  [Not refereed][Not invited]
     
    Protein tyrosine phosphorylation accompanies the integrin-mediated cell to substratum adhesion, and is essential for the progression of G1/S phase of the cell-cycle in normal fibroblasts. To examine how cellular protein tyrosine phosphatase (PTPase) activity is involved in regulating the adhesion-dependent protein tyrosine phosphorylation, we employed fibroblast cells bearing an active form of a protein tyrosine kinase (PTK), v-Src. We found that the v-Src induced tyrosine phosphorylation in certain proteins such as tensin, talin, p120, p80/85 (cortactin) and paxillin was greatly reduced when the cell to substratum adhesion was lost. Re-adhesion of the cells onto fibronectin restored these phosphorylation events, while this was inhibited by the addition of RGD peptide. The kinase activity of the v-Src was unchanged by the loss of cell to substratum adhesion. On the other hand, treatment with a protein tyrosine phosphatase inhibitor vanadate caused much the same increase in the v-Src-mediated cellular tyrosine phosphorylation between cells adhered to the culture environments and cells kept in suspension. These data suggest that PTPase(s) appears to be more critical than the v-Src PTK in determining the cell adhesion-dependent protein tyrosine phosphorylation. Moreover, most of the protein tyrosine phosphorylations that are mediated by the v-Src but still dependent on the cell adhesion were indeed greatly reduced during an anchorage-independent growth of v-Src cells. Thus our data collectively indicate that the v-Src induced high level of tyrosine phosphorylation in certain types of proteins are still under the control of the integrin(s) or the cell adhesion to culture substratum, and most of these adhesion-regulated high levels of tyrosine phosphorylations are not essential for the transformed phenotype.
  • 細胞接着とシグナル伝達 インテグリンを介した細胞基質間接着におけるシグナル伝達
    真崎 雄一, 佐邊 壽孝  組織培養工学  23-  (6)  218  -222  1997/05  [Not refereed][Not invited]
  • Y Mazaki, S Hashimoto, H Sabe  JOURNAL OF BIOLOGICAL CHEMISTRY  272-  (11)  7437  -7444  1997/03  [Not refereed][Not invited]
     
    The versatility of integrin functions is mediated by engagement of a number of proteins that assemble with integrins. Among them, paxillin is one of the important molecules interacting with a variety of signaling molecules and cytoskeletal building blocks. We report here that paxillin is not a single molecule with a unique physiological property. We identified two human paxillin isoforms, beta and gamma. These isoforms have distinct amino acid insertions; each consists of a distinct exon, at the same site of previously reported paxillin (paxillin alpha). Several proteins were co-precipitated with paxillin, and we found that beta bound to focal adhesion kinase but weakly to vinculin, and gamma bound to vinculin but only weakly to focal adhesion kinase, although both bound equally to talin. No additional proteins were found to bind to beta and gamma over those binding to alpha. Unlike the alpha isoform, beta and gamma mRNAs were not detected in normal tissues, but several cancer cells expressed both alpha and beta proteins simultaneously. All three isoform proteins were expressed in promonocytic cells with ratios comparable with each other, and the expression patterns were altered during differentiation of floating promonocytic cells into adherent macrophage-like cells. Therefore, each isoform of paxillin exhibits distinct expression and different biochemical as well as physiological properties and thereby appears to act as a distinct module involved in different functions of integrins.
  • Y Mazaki, S Hashimoto, H Sabe  JOURNAL OF BIOLOGICAL CHEMISTRY  272-  (11)  7437  -7444  1997/03  [Not refereed][Not invited]
     
    The versatility of integrin functions is mediated by engagement of a number of proteins that assemble with integrins. Among them, paxillin is one of the important molecules interacting with a variety of signaling molecules and cytoskeletal building blocks. We report here that paxillin is not a single molecule with a unique physiological property. We identified two human paxillin isoforms, beta and gamma. These isoforms have distinct amino acid insertions; each consists of a distinct exon, at the same site of previously reported paxillin (paxillin alpha). Several proteins were co-precipitated with paxillin, and we found that beta bound to focal adhesion kinase but weakly to vinculin, and gamma bound to vinculin but only weakly to focal adhesion kinase, although both bound equally to talin. No additional proteins were found to bind to beta and gamma over those binding to alpha. Unlike the alpha isoform, beta and gamma mRNAs were not detected in normal tissues, but several cancer cells expressed both alpha and beta proteins simultaneously. All three isoform proteins were expressed in promonocytic cells with ratios comparable with each other, and the expression patterns were altered during differentiation of floating promonocytic cells into adherent macrophage-like cells. Therefore, each isoform of paxillin exhibits distinct expression and different biochemical as well as physiological properties and thereby appears to act as a distinct module involved in different functions of integrins.
  • 細胞基質間接着シグナリング パキシリンの役割
    佐邊 壽孝  組織培養研究  16-  (1)  20  -20  1997/03  [Not refereed][Not invited]
  • 細胞接着とシグナル伝達
    蛋白質・核酸・酵素(共立出版)  1997  [Not refereed][Not invited]
  • 細胞基質間接着・脱接着のシグナル
    組織培養工学  23-  32  -36  1997  [Not refereed][Not invited]
  • T Tanaka, R Yamaguchi, H Sabe, K Sekiguchi, JM Healy  FEBS LETTERS  399-  (1-2)  53  -58  1996/12  [Not refereed][Not invited]
     
    Short cytoplasmic domains of integrin heterodimers are crucial for transduction of signals generated by adhesion of cells to the extracellular matrix. Here, we describe the use of peptides mimicking the intracellular tails of integrin alpha(5) beta(1) to assay in vitro associations with cytoskeletal proteins. Our results suggest that the focal adhesion protein, paxillin, may interact directly with the intracellular region of the integrin beta(1) subunit. Paxillin is known to form stable complexes with several signaling molecules, including focal adhesion kinase. Physical interaction between paxillin and the beta(1) cytoplasmic domain suggests a model in which paxillin may function as a key intermediary in integrin-mediated signal transduction.
  • MY Pu, AA Akhand, M Kato, M Hamaguchi, T Koike, H Iwata, H Sabe, H Suzuki, Nakashima, I  ONCOGENE  13-  (12)  2615  -2622  1996/12  [Not refereed][Not invited]
     
    The kinase activity of p60(c-src) has been shown to be basically regulated through phosphorylation and dephosphorylation of Y527. We found that catalytic activity of the immunoprecipitated c-Src kinase from NIH3T3 cells was elevated several folds by exposure to 0.5-50 mu M of sulfhydryl-reactive Hg2+. V-max of the kinase was increased whereas K-m was decreased. N-acetylcysteine neutralized this Hg2+ effect, suggesting a critical role of the Hg2+-mediated sulfhydryl modification of the kinase in the mechanism. Addition of protein tyrosine phosphatase inhibitor Na3VO4 into the reaction mixture did not inhibit the Hg2+-mediated activation. Further study revealed that Hg2+ was capable of activating the v-Src kinase lacking Y527 and the c-Src kinase from mutant cells defective of the Y527-phosphorylating Csk kinase. Cyanogen bromide cleavage maps of radiolabeled Src proteins showed that Hg2+ selectively promoted the autophosphorylation at Y416 and that the previously in vivo radiolabeled phosphorous on Y527 was not deleted during the promotion of Y416 autophosphorylation by Hg2+, Phosphoamino acid analysis demonstrated selective promotion of phosphorylation at tyrosine but not at serine/threonine. Not like bivalent Hg2+, monovalent p-chloromercuribenzenesulfonic acid was incapable of activating c-Src kinase. These results suggest a novel Y416 phosphorylation-linked activation pathway for Src kinases which is initially triggered independent of Y527-mediated or serine/threonine phosphorylation-linked regulation, possibly through sulfhydryl-based protein structural modification for functional alteration.
  • T Tanaka, R Yamaguchi, H Sabe, K Sekiguchi, JM Healy  FEBS LETTERS  399-  (1-2)  53  -58  1996/12  [Not refereed][Not invited]
     
    Short cytoplasmic domains of integrin heterodimers are crucial for transduction of signals generated by adhesion of cells to the extracellular matrix. Here, we describe the use of peptides mimicking the intracellular tails of integrin alpha(5) beta(1) to assay in vitro associations with cytoskeletal proteins. Our results suggest that the focal adhesion protein, paxillin, may interact directly with the intracellular region of the integrin beta(1) subunit. Paxillin is known to form stable complexes with several signaling molecules, including focal adhesion kinase. Physical interaction between paxillin and the beta(1) cytoplasmic domain suggests a model in which paxillin may function as a key intermediary in integrin-mediated signal transduction.
  • 細胞基質間接着研究の現状と今後の課題
    佐邊 壽孝  実験医学  14-  (17)  2312  -2316  1996/11  [Not refereed][Not invited]
  • 細胞基質接着斑蛋白質パキシリンは蛋白質結合性,発現特性が異なる複数のアイソフォームから成る
    佐邊 壽孝  日本癌学会総会記事  55回-  76  -76  1996/09  [Not refereed][Not invited]
  • 柳 茂, 菅原 斉, 佐邊 壽孝, 山村 博平, 黒崎 知博  日本分子生物学会年会プログラム・講演要旨集  19-  (0)  1996/08/01  [Not refereed][Not invited]
  • BOUGERET C, DELAUNAY T, ROMERO F, JULLIEN P, SABE H, HANAFUSA H, BENAROUS R, FISCHER S  The Jouanal of Biological Chemistry  271-  (13)  7465  -7472  1996  [Not refereed][Not invited]
  • YANAGI S, SUGAWARA H, KUROSAKI M, SABE H, YAMAMURA H, KUROSAKI T  J.Biol.Chem.  271-  (48)  30487  -30492  1996  [Not refereed][Not invited]
  • CSK Enhances Insulin-stimulated Dephosphorylation of focal adhesion proteins.(共著)
    Mol.Cell.Biol.  16-  4765  -4772  1996  [Not refereed][Not invited]
  • インテグリンを介した細胞基質間接着とシグナル伝達
    第107回日本医学会シンポジュウム記録集  6  -16  1996  [Not refereed][Not invited]
  • パキシリン、Fak
    "分子細胞生物学辞典"(東京化学同人)  1996  [Not refereed][Not invited]
  • "細胞接着とシグナリング" 編集(共著)
    実験医学増刊号  1996  [Not refereed][Not invited]
  • J.Biol.Chem.  271-  (48)  30487  -30492  1996  [Not refereed][Not invited]
  • CSK Enhances Insulin-stimulated Dephosphorylation of focal adhesion proteins.(共著)
    Mol.Cell.Biol.  16-  4765  -4772  1996  [Not refereed][Not invited]
  • v-Src癌化細胞における細胞基質間接着はv-Srcによる細胞内蛋白質チロシンリン酸化に関与する
    佐邊 壽孝  日本癌学会総会記事  54回-  158  -158  1995/09  [Not refereed][Not invited]
  • 基礎研究はがんを制圧できるか v-Crkによる細胞がん化 細胞基質接着斑タンパク質paxillinとv-Crk,Cskとの相互作用を中心として
    佐邊 壽孝  細胞工学  14-  (5)  506  -512  1995/05  [Not refereed][Not invited]
  • 蛋白-蛋白相互作用によるシグナルの受け渡し 細胞基質間接着,脱接着による細胞内シグナリング
    佐邊 壽孝  実験医学  13-  (6)  704  -711  1995/04  [Not refereed][Not invited]
  • SABE H, SHOELSON S E, HANAFUSA H  J.Biol.Chem.  270-  (52)  31219  -31224  1995  [Not refereed][Not invited]
  • v-Crkによる細胞癌化:細胞基質斑蛋白質paxillinとv-Crk, Cskとの相互作用を中心として
    細胞工学  14-  506  -512  1995  [Not refereed][Not invited]
  • 細胞基質間接着、脱接着による細胞内シグナリング
    シグナル伝達における分子間相互作用(竹縄忠臣、羊土社)実験医学増刊号  13-  704  -711  1995  [Not refereed][Not invited]
  • J.Biol.Chem.  270-  (52)  31219  -31224  1995  [Not refereed][Not invited]
  • Functional analysis of Csk in signal transduction through the B cell antigen receptor.(共著)
    Mol.Cell.Biol.  14-  7306  -7313  1994  [Not refereed][Not invited]
  • TAVOLONI N, INOUE H, SABE H, HANAFUSA H  J.Cell Biol.  126-  (2)  475  -483  1994  [Not refereed][Not invited]
  • Specific motifs recognized by the SH2 domains of Csk, 3BP2, fes/fps,Grb-2, SHPTP1, SHC, Syk and Vav.(共著)
    Mol.Cell.Biol.  14-  2777  -2785  1994  [Not refereed][Not invited]
  • Tyrosine kinase Lyn and Syk regulate B cell receptor-coupled calcium mobilization through distinct pathways.(共著)
    EMBO J.  13-  1341  -1349  1994  [Not refereed][Not invited]
  • Comparative study of three protein tyrosine phosphatases : ChPTP1 specifically dephosphorylates c-Src tyrosine 527.(共著)
    J.Biol.Chem.  269-  20194  -20200  1994  [Not refereed][Not invited]
  • IL-2 can support growth of CD8+ T cells but not CD4+ T cells of human IL-2 receptor β-chain transgenic mice.(共著)
    J.Immunol.  153-  5373  -5381  1994  [Not refereed][Not invited]
  • src ; temperature-sensitive mutant ; thymidine kinase
    分子生物学・免疫学キ-ワ-ド辞典(永田和宏、長野孝、宮坂信行、宮坂昌之編、医学書院)  1994  [Not refereed][Not invited]
  • Functional analysis of Csk in signal transduction through the B cell antigen receptor.(共著)
    Mol.Cell.Biol.  14-  7306  -7313  1994  [Not refereed][Not invited]
  • Specific motifs recognized by the SH2 domains of Csk, 3BP2, fes/fps,Grb-2, SHPTP1, SHC, Syk and Vav.(共著)
    Mol.Cell.Biol.  14-  2777  -2785  1994  [Not refereed][Not invited]
  • Tyrosine kinase Lyn and Syk regulate B cell receptor-coupled calcium mobilization through distinct pathways.(共著)
    EMBO J.  13-  1341  -1349  1994  [Not refereed][Not invited]
  • Comparative study of three protein tyrosine phosphatases : ChPTP1 specifically dephosphorylates c-Src tyrosine 527.(共著)
    J.Biol.Chem.  269-  20194  -20200  1994  [Not refereed][Not invited]
  • IL-2 can support growth of CD8+ T cells but not CD4+ T cells of human IL-2 receptor β-chain transgenic mice.(共著)
    J.Immunol.  153-  5373  -5381  1994  [Not refereed][Not invited]
  • SH2領域を介したCskによるc-Srcの活性抑制機構
    Medical Immunology(国際医書出版)  26-  193  -196  1993  [Not refereed][Not invited]
  • SABE H, KNUDSEN B, OKADA M, NADA S, NAKAGAWA H, HANAFUSA H  Proc.Natl.Acad.Sci.USA  89-  (6)  2190  -2194  1992  [Not refereed][Not invited]
  • Proc.Natl.Acad.Sci.USA  89-  (6)  2190  -2194  1992  [Not refereed][Not invited]
  • 浅野 正岳, 石田 靖雅, 佐辺 寿孝, 岡崎 佐江子, 本庶 佑, 日合 弘  日本疾患モデル学会記録  8-  (0)  61  -61  1992  [Not refereed][Not invited]
  • Biochemical evidence for a third chain of the interleukin-2 receptor.(共著)
    J.Biol.Chem.  266-  22186  -22191  1991  [Not refereed][Not invited]
  • SABE H, KUNO J, KOROMILAS A, SAITO Y, KINASHI T, UEDA M, TAKAMATSU T, HAMAGUCHI M, HONJO T  Inter.Immunol.  3-  (11)  1137  -1148  1991  [Not refereed][Not invited]
  • Establishment of an IL2 dependent T cell Line from a patient with a multiple seclerosis, which express unregulated IL2 receptor and has clonal abnormality 21 trisomy.(共著)
    Int.J.Immunolotherapy  8-  (3)  123  -131  1991  [Not refereed][Not invited]
  • Biochemical evidence for a third chain of the interleukin-2 receptor.(共著)
    J.Biol.Chem.  266-  22186  -22191  1991  [Not refereed][Not invited]
  • Inter.Immunol.  3-  (11)  1137  -1148  1991  [Not refereed][Not invited]
  • Establishment of an IL2 dependent T cell Line from a patient with a multiple seclerosis, which express unregulated IL2 receptor and has clonal abnormality 21 trisomy.(共著)
    Int.J.Immunolotherapy  8-  (3)  123  -131  1991  [Not refereed][Not invited]
  • Stepwise formation of the high-affinity complex of the interleukin 2 receptor.(共著)
    Inter.Immunol.  2-  1165  -1177  1990  [Not refereed][Not invited]
  • Effecient transient expression of cDNA in CTLL-2 cells expressing polyoma large-antigen.(共著)
    Technique  2-  189  -193  1990  [Not refereed][Not invited]
  • HTLV-1 p27rex stabilaizes human interleukin 2 receptor a chain m RNA.(共著)
    EMBO J.  9-  4164  -4166  1990  [Not refereed][Not invited]
  • Stepwise formation of the high-affinity complex of the interleukin 2 receptor.(共著)
    Inter.Immunol.  2-  1165  -1177  1990  [Not refereed][Not invited]
  • Effecient transient expression of cDNA in CTLL-2 cells expressing polyoma large-antigen.(共著)
    Technique  2-  189  -193  1990  [Not refereed][Not invited]
  • HTLV-1 p27rex stabilaizes human interleukin 2 receptor a chain m RNA.(共著)
    EMBO J.  9-  4164  -4166  1990  [Not refereed][Not invited]
  • SAITO Y, SABE H, SUZUKI N, KONDO S, OGURA T, SHIMIZU A, HONJO T  J.Exp.Med.  168-  (5)  1563  -1572  1988  [Not refereed][Not invited]
  • Molecular analysis of the formation of the high affinity interlsukin 2 receptor.(共著)
    Mol.Biol.Med.  5-  123  -138  1988  [Not refereed][Not invited]
  • Differential effects on interleukin 2 receptors (p75 and p55) by the pXgene of HTLV-(]G0001[) virus.(共著)
    Inter.J.Cancer  41-  880  -885  1988  [Not refereed][Not invited]
  • Molecular analysis of the formation of the high affinity interlsukin 2 receptor.(共著)
    Mol.Biol.Med.  5-  123  -138  1988  [Not refereed][Not invited]
  • Differential effects on interleukin 2 receptors (p75 and p55) by the pXgene of HTLV-(]G0001[) virus.(共著)
    Inter.J.Cancer  41-  880  -885  1988  [Not refereed][Not invited]
  • Kazuhiko Tsuboi, Kazuhiko Tsuboi, Kazunori Hirayoshi, Kaoru Takeuchi, Hisataka Sabe, Yutaka Shimada, Gakuji Ohshio, Takayoshi Tobe, Masakazu Hatanaka  Biochemical and Biophysical Research Communications  146-  (2)  699  -704  1987/07/31  [Not refereed][Not invited]
     
    We have examined the level of the c-myc transcript in 6 esophageal, 16 gastric, 19 colorectal and 1 anal cancer tissue samples; these included four lymph nodes and six hepatic metastases obtained surgically. The esophageal cancer tissues were without an increase of the c-myc transcript, some of the gastric cancer samples showed a two to three fold increase and most of the colorectal and the one anal cancer samples showed a two to ten fold increase when compared with a normal mucosal layer. Therefore, the level of the c-myc transcript in human gastrointestinal malignacies shows organ dependency. Local, lymphatic, and hepatic metastases showed little difference in the level of c-myc mRNA from that of the primary tumor. © 1987.
  • Molecular genetic studies on the interleukin 2 receptor.(共著)
    Recent Advance in Leukemia and Lymphoma  309  -312  1987  [Not refereed][Not invited]
  • KONDO S, KINOSHITA M, SHIMIZU A, SAITO Y, KONISHI M, SABE H, HONJO T  Nature  327-  (6117)  64  -67  1987  [Not refereed][Not invited]
  • Expression of a provirus of human T cell leukemia virus type (]G0001[) by DNA transfection.(共著)
    J.Gen.Virol.  68-  499  -506  1987  [Not refereed][Not invited]
  • Molecular genetic studies on the interleukin 2 receptor.(共著)
    Recent Advance in Leukemia and Lymphoma  309  -312  1987  [Not refereed][Not invited]
  • Nature  327-  (6117)  64  -67  1987  [Not refereed][Not invited]
  • Expression of a provirus of human T cell leukemia virus type (]G0001[) by DNA transfection.(共著)
    J.Gen.Virol.  68-  499  -506  1987  [Not refereed][Not invited]
  • STRUCTURE AND FUNCTION OF THE INTERLEUKIN-2 RECEPTOR - AFFINITY CONVERSION MODEL
    A SHIMIZU, S KONDO, H SABE, N ISHIDA, T HONJO  IMMUNOLOGICAL REVIEWS  92-  103  -120  1986  [Not refereed][Not invited]
  • Genomic expression of human T-lymphotropic virus (HTLV-(]G0001[)).(共著)
    AIDS Res.  2-  s79-s85  1986  [Not refereed][Not invited]
  • Structure analysis of p28 adult T cell leukemia-associated antigen.(共著)
    J.Gen.Virol.  67-  1373  -1379  1986  [Not refereed][Not invited]
  • STRUCTURE AND FUNCTION OF THE INTERLEUKIN-2 RECEPTOR - AFFINITY CONVERSION MODEL
    A SHIMIZU, S KONDO, H SABE, N ISHIDA, T HONJO  IMMUNOLOGICAL REVIEWS  92-  103  -120  1986  [Not refereed][Not invited]
  • Genomic expression of human T-lymphotropic virus (HTLV-(]G0001[)).(共著)
    AIDS Res.  2-  s79-s85  1986  [Not refereed][Not invited]
  • Structure analysis of p28 adult T cell leukemia-associated antigen.(共著)
    J.Gen.Virol.  67-  1373  -1379  1986  [Not refereed][Not invited]
  • N ISHIDA, H KANAMORI, T NOMA, T NIKAIDO, H SABE, N SUZUKI, A SHIMIZU, T HONJO  NUCLEIC ACIDS RESEARCH  13-  (21)  7579  -7589  1985  [Not refereed][Not invited]
  • A SHIMUZU, S KONDO, S TAKEDA, J YODOI, N ISHIDA, H SABE, H OSAWA, T DIAMANTSTEIN, T NIKAIDO, T HONJO  NUCLEIC ACIDS RESEARCH  13-  (5)  1505  -1516  1985  [Not refereed][Not invited]
  • IZUI K, MIWA T, KAJITANI M, FUJITA N, SABE H, ISHIHAMA A, KATSUKI H  Nucleic Acids Res.  13-  (1)  59  -71  1985  [Not refereed][Not invited]
  • Nucleic Acids Res.  13-  7579  -7589  1985  [Not refereed][Not invited]
  • A SHIMUZU, S KONDO, S TAKEDA, J YODOI, N ISHIDA, H SABE, H OSAWA, T DIAMANTSTEIN, T NIKAIDO, T HONJO  NUCLEIC ACIDS RESEARCH  13-  (5)  1505  -1516  1985  [Not refereed][Not invited]
  • Nucleic Acids Res.  13-  (1)  59  -71  1985  [Not refereed][Not invited]
  • GENOMIC STRUCTURE OF HTLV (HUMAN T-CELL LEUKEMIA-VIRUS) - DETECTION OF DEFECTIVE GENOME AND ITS AMPLIFICATION IN MT-2 CELLS
    N KOBAYASHI, H KONISHI, H SABE, K SHIGESADA, T NOMA, T HONJO, M HATANAKA  EMBO JOURNAL  3-  (6)  1339  -1343  1984  [Not refereed][Not invited]
  • NIKAIDO T, SHIMIZU A, ISHIDA N, SABE H, TESHIGAWARA K, MAEDA M, UCHIYAMA T, YODOI J, HONJO T  Nature  311-  (5987)  631  -635  1984  [Not refereed][Not invited]
  • GENOMIC STRUCTURE OF HTLV (HUMAN T-CELL LEUKEMIA-VIRUS) - DETECTION OF DEFECTIVE GENOME AND ITS AMPLIFICATION IN MT-2 CELLS
    N KOBAYASHI, H KONISHI, H SABE, K SHIGESADA, T NOMA, T HONJO, M HATANAKA  EMBO JOURNAL  3-  (6)  1339  -1343  1984  [Not refereed][Not invited]
  • Nature  311-  (5987)  631  -635  1984  [Not refereed][Not invited]
  • J.Gen.Appl.Microbiol  30-  27  -33  1984  [Not refereed][Not invited]
  • Gene  31-  (1/3)  279  -283  1984  [Not refereed][Not invited]
  • Primary structure of phosphoenolpyruvate carboxylase in E.coli.(共著)
    Proceedings of the 34th Meeting on Protein Structure  105  -108  1983  [Not refereed][Not invited]
  • FEBS Letters  133-  (2)  311  -315  1981  [Not refereed][Not invited]
  • EPB41L5 is essential for post-transcriptional regulation of cadherin and integrin in epithelial-mesenchymal transition during mouse gastrulation.
    M. Hirano, S. Hashimoto, S. Yonemura, H. Sabe, S. Aizawa  J. Cell Biol. (in press).  [Not refereed][Not invited]
  • J. Biochem. Minireview(in press)  [Not refereed][Not invited]
  • Paxillin-associated ArfGAPs-their isoform specificities and roles in coordination-in
    The ARF Book  [Not refereed][Not invited]
  • J. Biochem. Minireview(in press)  [Not refereed][Not invited]
  • Paxillin-associated ArfGAPs-their isoform specificities and roles in coordination-in
    The ARF Book  [Not refereed][Not invited]

Industrial Property Rights

Research Grants & Projects

  • 文部科学省:科学研究費補助金(基盤研究(B))
    Date (from‐to) : 2014 -2016 
    Author : 佐邊 壽孝
  • 文部科学省:科学研究費補助金(新学術領域研究(研究領域提案型))
    Date (from‐to) : 2011 -2015 
    Author : 佐邊 壽孝, 小根山 千歳
     
    これ迄に、Arf6を中心とした細胞内シグナル経路が多くの乳癌の浸潤転移に関与していることを明らかにして来た。その際、Arf6はGEP100によって活性化され、活性化したArf6はAMAP1をエフェクターとする。また、GEP100はEGFR,Her2等チロシンリン酸化酵素型受容体により活性化される。一方、乳癌患者標本のコホート解析から、TGFβ1が、乳癌細胞の癌幹細胞様細胞化と浸潤転移性獲得に最も高頻度で関与する因子である事が示されている。TGFβ1によってもこのArf6経路が活性化すること、その際、TGFβ1によってチロシンリン酸化酵素型受容体であるc-Metが活性化されること、活性化されたc-MetにGEPIOOが結合していることを見いだしている。この結合は直接ではなく、Gab1とよばれるadaptorが介在した。本年度は、TGFβ1によるc-Metの活性化機構を明らかにすべく解析を進めた。癌の約半数にp53遺伝子の変異が認められ、また、このような変異の多くはgain-of functionである。解析の結果、特定の乳癌細胞におけるTGFβ1によるArf6の活性化には、p53のgain-of-function変異が必須である事を明らかにした。変異p53の発現をsiRNA法により抑制すると、TGFβ1によるArf6活性化は抑制され、浸潤の活性化もなくなった。野生型p53の戻し実験系においても、TGFβ1によるArf6の活性化はなかった。また、野生型p53の戻すとArf6経路の特定の蛋白質の発現が抑制されること、この抑制は当該mRNAの転写後の制御であることを見出している。また、Srcファミリーキナーゼの活性化によるmicroRNAを介したILKの発現亢進が細胞接着斑の形成の破綻を引き起こしがん悪性化に繋がることを見出した。
  • Ministry of Education, Culture, Sports, Science and Technology:Grants-in-Aid for Scientific Research(基盤研究(A))
    Date (from‐to) : 2008 -2011 
    Author : Hisataka SABE
     
    (抄録なし)
  • Ministry of Education, Culture, Sports, Science and Technology:Grants-in-Aid for Scientific Research(特定領域研究)
    Date (from‐to) : 2005 -2009 
    Author : Hisataka SABE
     
    Most malignant human tumors are epithelial cell origin. The basic fear of such tumors are based on their invasion and metastasis. Activation of integrins and inactivation of E-cadherin are hallmark characteristics for such invasive and metastatic properties. Here our purpose is to understanding the underlying molecular mechanisms for integrin activation and E-cadherin inactivation in the malignant development of tumor cells.
  • Ministry of Education, Culture, Sports, Science and Technology:Grants-in-Aid for Scientific Research(基盤研究(B))
    Date (from‐to) : 2006 -2007 
    Author : Hisataka SABE
     
    We aim to elucidate mechanisms controlling cell migration and the polarity formation, via investigating their relationship to intracellular vesicle trafficking. Our other main research interest is to understand the principal mechanisms involved in maintenance of epithelial tissue integrity, as well as cancer cell invasion and metastasis which occurs as a result of the disruption of the epithelial integrity. Although biology is a diverse subject, we are constantly aiming towards understanding whether a common fundamental mechanism actually exists behind the complicated phenomena of living or...
  • Ministry of Education, Culture, Sports, Science and Technology:Grants-in-Aid for Scientific Research(基盤研究(B))
    Date (from‐to) : 2004 -2005 
    Author : Hisataka SABE, 真崎 雄一, 橋本 茂, 三浦 浩一, 橋本 あり
     
    Cell migration is a multifactorial process in which a number of distinct events occur simultaneously. The major purpose of this study is to understand basic molecules and mechanisms coordinately regulating cell migration and invasion.Arf6 plays essential roles in recycling of plasma membrane component, as well as both membrane and cytoskeletal remodeling at cell peripheries. We have previously identified several proteins bearing ArfGAP domains as binding proteins to paxillin, an integrin signaling adaptor/scaffolding protein. These ArfGAPs include AMAP1 and AMAP2. We have also shown that AM...
  • 文部科学省:科学研究費補助金(特定領域研究(C), 特定領域研究)
    Date (from‐to) : 2000 -2004 
    Author : 佐邊 壽孝, 広橋 説雄, 永渕 昭良
     
    がん細胞、特に上皮由来癌の浸潤機序とそれに関わる分子機構を明らかにするため、インテグリン裏打ち蛋白質とカドヘリン裏打ち蛋白質に着目し解析を進めている。これまでの研究を踏まえ本年度は、パキシリンとその結合性ArfGAPを含む、乳癌の浸潤活性に重要な蛋白質複合体を同定し、その存在が乳癌(細胞株、病理切片)の浸潤形質と良く一致すること、siRNA法による個々の蛋白質発現の阻害や蛋白質複合体の形成をペプチド等によって阻害することによって乳癌の浸潤と転移を著しく阻害できることを示し、この蛋白質複合体が抗癌剤開発の分子標的としての評価対象になり得ることを提示した(佐邊)。一方、懸案であるEカドヘリンとNカドヘリンとの動態の差異がどのようにして癌の浸潤形質獲得と連関するのかに関して、カドヘリンの動態の差異と癌の浸潤活性の両方にArf6が関与することを明らかにし、細胞間接着形成においてArf6の活性制御に関わる細胞表面受容体も同定した。分子機序に関しては現在解析中である(佐邊)。カドヘリン裏打ち蛋白質に関しては、F9細胞を用いて細胞の接着形態における役割を検討し、β-カテニンとプラコグロビン両者を欠損させると細胞接着能が減弱することを示した(永渕)。
  • Ministry of Education, Culture, Sports, Science and Technology:Grants-in-Aid for Scientific Research(基盤研究(B))
    Date (from‐to) : 2002 -2003 
    Author : Hisataka SABE, 近藤 明子, 八木 良平, 矢野 元, 橋本 茂, 橋本 あり
     
    Previously we reported that AMAP2/PAG3/Papα/KIAAO400, a GTPase-activating protein (GAP), acts to antagonize Arf6 function when overexpressed, while it was shown to exhibit efficient GAPing activities for other Arf isoforms in vitro. During this period of the two fiscal years we first found that AMAP2, through its ArfGAP domain, binds to GTP-Arf6, but not to GDP-Arf6 nor other Arfs irrespective of nucleotide status. The majority of AMAP2 was localized to intracellular tubulovesicular structures, and redistributed to Arf6-enriched membrane areas upon Arf6 activation. In HeLa cells, Arf6 has b...
  • 文部科学省:科学研究費補助金(萌芽的研究, 萌芽研究)
    Date (from‐to) : 2001 -2002 
    Author : 佐邊 壽孝
     
    これまでにインテグリンを介する細胞の接着と運動制御において、paxillinとp130Casのチロシンリン酸化が重要な役割を果たすこと、その内paxillinのTyr31/118のリン酸化が上皮細胞の運動先端部におけるRhoAの活性抑制に関与してしていることを明らかにしてきた。今回、paxillinのTyr40/181のリン酸化が、運動する上皮細胞においてCdc42とRac1の活性制御に深く関与することを明らかにし、そのことによりpaxillinが細胞運動極性の制御に深く関わることを示した。即ち、これまでの我々の知見(JBC,275,27155(2000))に加え、今回FRET法(阪大、微研、松田道行先生との共同研究)での評価も行った結果、Tyr40/181のリン酸化変異体を発現させた上皮細胞では、運動中にCdc42とRac1の正常を逸脱した活性化が起っていることを明らかにすることができた異常な活性化が起る部位とその方向への運動極性のぶれは概ね一致していた。Tyr31/118の変異体ではこのようなことは観察されなかった。この異常な活性化は細胞の局所でのみ観察されることであり、細胞全体のCdc42やRaclの活性量を生化学的に測定しても、コントロールと比べ有為な変化を補足することはできなかった。現在、Cdc42やRac1の活性制御に到るTyr40/181のリン酸化の下流シグナル...
  • Ministry of Education, Culture, Sports, Science and Technology:Grants-in-Aid for Scientific Research(基盤研究(B))
    Date (from‐to) : 2000 -2001 
    Author : Hisataka SABE, 真崎 雄一, 矢野 元, 橋本 茂, 八木 良平
     
    ARF6 regulates endosomal recycling. We have shown that PAG3/Papα/KIAA0400 acts as a GTPase-activation protein (GAP) specific for ARF6.We study here molecular mechanims how PAG3 is involved in endosomal recycling to be an ARF6GAP. We found that PAG3, via its proline-rich region, binds to the src homology 3 (SH3) domain of several components of the endocytic machinery, and analysed its interaction with amphiphysin IIa. PAG3 existed at ARF6(Q67L)-positive membrane ruffles colocalized with amphyphysin Ha, but the majority exists at intracellular tubulovesicular structure. Overexpression of the ...
  • Ministry of Education, Culture, Sports, Science and Technology:Grants-in-Aid for Scientific Research(基盤研究(B))
    Date (from‐to) : 1998 -2001 
    Author : Hisataka SABE, 真崎 雄一, 矢野 元, 橋本 茂, 真崎 雄一, 矢野 元, 橋本 茂
     
    Temporal and spatial regulation of actin-based cytoskeletal organization and focal adhesion formation play an essential role in cell migration. We found that tyrosine phosphorylation of paxillin and pi3OCas was a prominent event upon integrin activation during epithelial-mesenchymal transdifferentiation and cell migration. Tyrosine phosphorylation of p130^ has been demonstrated to facilitate cell migration. We showed that tyrosine phosphorylation of paxillin cc acts to reduce haptotactic cell migration as well as transcellular invasive activities in several different experimental cell ...
  • 文部科学省:科学研究費補助金(特定領域研究(A))
    Date (from‐to) : 1999 -1999 
    Author : 佐邊 壽孝, 真崎 雄一, 矢野 元, 橋本 茂, Marius Sudol, Donald E, Joan S
     
    細胞が運動する際には、細胞の前方部にインテグリン接着点が新たに形成される。インテグリンは多くの種類の蛋白質をその細胞質領域に集積させることにより機能する。我々は、「このような裏打ち蛋白質群を集積させる過程が単に細胞質における自由拡散過程なのか、それとも、何か能動的な機序が存在するのか」との設問を立て、解析を開始した。それまで機能解析をしてきたパキシリンというインテグリン裏打ち蛋白質に関して上記のような解析をしたところ、このものは、核周辺領域に細胞質プールが存在し、運動中に細胞前方に形成されるラミニポデアにパキシリンを集積させる過程は、細胞質での自由拡散ではなく、何らかの能動的な機序が存在することを強く示唆する結果を得た。そこで、パキシリンをプローブとしパキシリンの細胞内動態を説明できるような蛋白質を検索したところ、小胞/膜/蛋白質の輸送に関与する低分子性GTP結合蛋白質であるARF蛋白質に対するGAP(GTPase-activating protein)活性を持つ一連の興味深い一連の蛋白質が得られ、これらの蛋白質をPAGs(Paxillin-associated ARFGAP proteins)と命名した。現在、5種類のcDNAが得られており、その内、2種に関してほぼ初期段階の解析が終了し、論文に纏めている。ARFは哺乳類では6種のアイソフォームが存在するが、解析の終わった2...
  • 文部科学省:科学研究費補助金(特定領域研究(A))
    Date (from‐to) : 1999 -1999 
    Author : 佐邊 壽孝, 佐藤 陽, 中村 邦明, 矢野 元
     
    上皮間充織形質転換、及び、細胞運動におけるインテグリン活性化に伴う細胞内の生化学的変化を調べた結果、インテグリン裏打ち蛋白質であるパキシリンとp130Casとのチロシンリン酸化の亢進が顕著な変化であることが分かった。パキシリンの全ての主要なリン酸化部位(4箇所)の同定は先年度に終了している。今年度は、パキシリンとp130Casとの機能とを対比させながら、それらのチロシンリン酸化の生理的役割を検討した。その結果、パキシリンは、上皮細胞の運動性に対して抑制的に作用することを見い出した。一方、この研究を行っている間に、p130Casのチロシンリン酸化は細胞運動性亢進に寄与することが報告され、我々もこの点は追試確認した。細胞の増殖形態に関して検討した結果、両者は級数的増殖速度には特に影響を及ぼさないが、細胞の飽和密度に顕著な影響を及ぼした。即ち、パキシリンは飽和密度を低下させ、p130Casは飽和密度を上昇させる作用を示した。これらの作用にも、それぞれのチロシンリン酸化が関与することが示唆された。従って、両者はチロシンリン酸化を介して、互いに見かけ上逆の効果を及ぼす可能性が考えられる。P130Casのチロシンリン酸化にはCrkやNckなどが結合し下流のシグナルを発生することが示されているが、パキシリンに関して検討した結果、これらの関与に関して否定的な結果を得、パキシリンチロシンリン酸...
  • 文部科学省:科学研究費補助金(特定領域研究(A))
    Date (from‐to) : 1999 -1999 
    Author : 佐邊 壽孝, 橋本 茂
     
    成熟単球cDNA libraryをパキシリンをプロープとしfar-western法にてスクリーニングし、パキシリン結合性蛋白質cDNAを単離同定した。その結果、ARFGAPを示す新規蛋白質が得られた。我々は、成熟単球細胞以外でもパキシリン結合性蛋白質の精製やfar-western法による同定を行っているが、同様にARFGAP活性示す別の蛋白質が得られている。我々は、これら一群の蛋白質をPAGs(Paxillin-associated ARFGAP proteins)と命名し、今回成熟単球から得られたものをPAG3と命名した。我々は、単球の成熟に伴ってパキシリンが発現誘導されチロシンリン酸化されることを明らかにしているが、PAG3もこの過程で発現誘導、チロシンリン酸化され、成熟単球において、主に細胞形質膜辺縁部にパキシリンを共局在した。ARFは哺乳類では6種のアイソフォームが存在し、小胞/膜/蛋白質の輸送やアクチン細胞骨格の再構成に関与することが知られている。PAG3はARF6に対するGAP(GTPase-activating protein)として機能することを明らかにすると共に、その強制発現は、パキシリンの細胞接着点への集積と細胞運動性とに対し、阻害的に働くことを明らかにした。さらに、PAG3はヒト動脈硬化病変における泡沫細胞に高レベル発現している可能性を示唆する観察結果も...
  • 文部科学省:科学研究費補助金(特定領域研究(A))
    Date (from‐to) : 1999 -1999 
    Author : 橋本 茂, 真崎 雄一, 佐邊 壽孝
     
    上皮系細胞の生存性維持には増殖因子やサイトカインの刺激に加えて細胞接着シグナルが必要である。本研究では、インテグリン接着を介するシグナル伝達の機能的制御機構についてインテグリン裏打ちタンパク質パキシリンに着目した解析を進めている。今年度は、インテグリン接着点へのパキシリンの集積機構について解析を進めた。これまでに、パキシリンには核周辺領域に細胞質プールが存在し、運動中に細胞前方に能動的に集積させる機構が存在することを示唆させる結果を得ていた。この集積機構に関わる分子を解析するために、パキシリンに結合する分子群を同定し、想定される機能を持つ分子の検索を行った。その結果、ARF GAPモチーフを持つ一群のタンパク質を見い出した。その中の一つであるPAG3(Paxillin-associated ARF GAP protein 3)は、マクロファージ様に分化したU937細胞のcDNAからパキシリンをプローブとしたファーウエスタン法により見い出した。PAG3は、未分化の単球では細胞質に存在しているが、単球が接着性を獲得すると発現が亢進し、細胞辺縁部へ局在し、パキシリンと共局在することが観察された。また、PAG3のARF GAP活性がパキシリンの接着点への局在に重要な役割を果たしていること、分化した単球細胞の運動性の制御に関わっていることを示唆させる結果を得た。従って、パキシリンの細胞...
  • 文部科学省:科学研究費補助金(特定領域研究(A))
    Date (from‐to) : 1998 -1998 
    Author : 左邊 壽孝, 矢野 元, 橋本 茂
     
    上皮細胞や内皮細胞、さらには分化した単球/マクロファージ細胞等の接着性細胞にとって、細胞接着はその生存性の維持に必須である。接着が維持されないと、これらの細胞はanoikisと呼ばれる、速やかなprogrammed ce11 deathに陥る。インテグリンの活性化以降、phosphatidyl3kinase(PI3K)からAkt/protein kinaseBを経て、最終的にはcaspaseの活性抑制に至る経路が現在提唱されるに至っている。しかし、肝心のインテグリンの直下因子でありP13Kの活性化を引き起こす蛋白質因子は同定されていない。以前には、これはFocal adhesion kinaseであると提唱されていたが、我々が幾つかの検討を行ったところ、Focal adhesion kinaseではないことが判明した。さらに検討を進めたところ、Focal adhesion kinaseと似た分子量(120-140kDa)のチロシンリン酸化蛋白質がインテグリンの活性化に伴ってPI3KのSH2(N)領域に結合することが明らかになった。同様な分子量のPI3K結合性蛋白質は、調べた限りanoikisを起こす全ての細胞に認められた。既知の、分子量が120-140kDaのチロシンリン酸化蛋白質に対する抗体で、入手可能なものすべてを検討したが、それらのいずれでもなかった。そこで、現在この蛋...
  • 文部科学省:科学研究費補助金(萌芽的研究)
    Date (from‐to) : 1998 -1998 
    Author : 佐邊 壽孝, 真崎 雄一
     
    インテグリンはその細胞質領域において多種類の蛋白質因子が集積し、この複合体を介してインテグリンのシグナル伝達は行われる。また、これらの複合体を介して細胞内の状況がインテグリンに伝わり、その結合性等が制御される。インテグリンと細胞外基質との接着は細胞の動的な運動過程において形成されるものであり、従って、このような動的過程においてどのようにしてインテグリンはそのシグナル伝達因子である集積蛋白質群と会合するのかは、細胞の運動性制御を明らかにする上で重要なポイントである。我々は、インテグリン裏打ち蛋白質パキシリンが細胞質においては、一部ゴルジ装置とオーバーラップする核周辺域の膜構造体に局在していることを明らかにした。パキシリンは可溶性蛋白質であることから、引き続き、パキシリンに結合する蛋白質の精製、単離同定を行ったところ、3種の新規蛋白質を見い出した。興味深いことにこれらは全てArfGAPに相当する配列を有しており、実際、in vitroではそのような活性を示すと考えられた。現在、細胞運動におけるパキシリンの細胞内輸送の分子機序を解明すべくその機能解析を進めている。
  • 文部科学省:科学研究費補助金(重点領域研究)
    Date (from‐to) : 1997 -1997 
    Author : 佐邊 嘉孝
     
    癌細胞の浸潤・転移過程は主として、インテグリンを介する細胞接着と細胞運動により担われている。細胞接着性や細胞運動性制御の基本機構を解明する一つのアプローチとして、インテグリンの裏打ち蛋白質であるパキシリンを分子基盤とした解析を進めている本研究においては、パキシリンの機能をさらに理解するため、細胞内での局在を検討し、それが細胞接着斑のみならず、細胞辺縁の活性部位、並びに、ゴルジ装置に存在することを示した。パキシリン自身はゴルジ装置に保持されるべき領域を持たない。我々は新規パキシリン結合性蛋白質を幾つか単離し解析しているが、その内の一つがゴルジ装置局在配列と考えられている配列を持っていることを見い出した。この蛋白質は細胞核周辺領域と細胞辺縁の活性部位に存在することも見い出している。現在、その機能を詳細に解析しており、細胞運動の基本機構を担う、パキシリンを始めとするインテグリン裏打ち蛋白質郡の細胞膜直下への輪送と集積の分子機構の解明へと研究を進めている。
  • 文部科学省:科学研究費補助金(重点領域研究)
    Date (from‐to) : 1997 -1997 
    Author : 佐邉 壽孝
     
    単球細胞の分化・成熟過程、血管内皮細胞への接着を介する炎症部位への浸潤は、癌細胞の浸潤・転移過程を解析するモデル系として注目されている。また、それ自身、粥状動脈硬化の発症機序の解析とも相まって注目される。これらの過程には、インテグリンを介する細胞接着が重要な役割を果たす。我々は、インテグリンジグナル伝達に、インテグリン裏打ち蛋白質の一つであるパキシリンが非常に重要な役割を果たすことを示し、さらに最近、単球芽細胞及び癌細胞に強く発現する新規アイソフォーム(β,γ)を見い出している。本研究では、まず、β,γアイソフォーム特異的やモノクローナル抗体作成を行い、抗体を用いて、組織におけるそれらの発現を検討した。その結果、これらのアイソフォームはαアイソフォームとは異なり、正常や組織には殆ど発現していないが、発生途上の神経堤細胞や、運動性の高い細胞に発現していることを明らかにした。γアイソフォーム遺伝子エキソンはヒトに存在しマウスには存在しないことを明らかにし、さらに、単球芽細胞の成熟に伴いその発現量は増加するが、γアイソフォームに特異的に結合する蛋白質を成熟単球細胞抽出液において検出することに成功し、現在、その分子クローニングを進めている。
  • 文部科学省:科学研究費補助金(重点領域研究)
    Date (from‐to) : 1996 -1996 
    Author : 左邊 壽孝
     
    癌細胞の悪性増殖へ深く関わる細胞基質間接着制御機構及びそのシグナル伝達に関し、以下の諸点を明らかにした。1。細胞基質間接着に伴って,tansin,Fak,paxillin等のインテグリン裏打ち蛋白質群のチロシンリン酸化が誘起されるが、この反応の制御には、リン酸化酵素よりも、脱リン酸化酵素が主体であることを示す結果を得た。この脱リン酸化酵素の見かけ上の活性は細胞基質間接着及び接着に伴うアクチンストレスファイバーのintegrityに制御されていると考えられる。2。ヒトpaxillin isoformを2種単離し、これらが或種の癌細胞や単球細胞に発現していることを明らかにした。これらの新規isoformは単独で発現しているのではなく、同一細胞に複数のisoformが発現しており、各々異なった機能を持つことが明らかになった。3。g isoformは細胞運動性に関係していることを示唆する結果を得ている。4。paxillin isoformの更なる機能解析を行う為、マウスの遺伝子座位のcloningを行い、その構造解析を終了した。g isoformの遺伝子座位に関しては、マウスとヒトとで大きく異なる。5。サイトカイン刺激がインテグリン裏打ち蛋白質群のチロシンリン酸化に作用し、細胞の形態・運動性を変化させることが知られているが、モデル系として、インシュリンを用い、インシュリン刺激によるイ...
  • Ministry of Education, Culture, Sports, Science and Technology:Grants-in-Aid for Scientific Research(国際学術研究)
    Date (from‐to) : 1995 -1996 
    Author : 淀井 淳司, Junji YODOI, コルスマイヤー スタンレ, チュルツ トマス, キン ユン, キャンビア ジョン, 花房 秀三郎, カリン マイケル, 口野 嘉幸, 豊國 伸哉, 山本 健一, 佐邊 壽孝, チェー ホーズン, パク サムチュル, ストルツ ギゼラ, チュルソ トーマス, チュルツ トーマス
     
    Human thioredoxin (TRX) has a potent thiol reducing activity and plays an important role on the redox regulation. Yodoi and the the members of the international cooperation program of the redox regulation and cellular signal transduction demonstrated the following results : 1) the targeted disruption of the TRX was embryonic lethal, suggesting the essential role of TRX in the mouse embryogenesis and differentiation. 2) the oxidative responsive element in the thioredoxin promoter was identified. 3) TRX and Ref-1 which is important for the activation of the AP-1 was demonstrated to be able to...
  • Ministry of Education, Culture, Sports, Science and Technology:Grants-in-Aid for Scientific Research(総合研究(A), 基盤研究(A))
    Date (from‐to) : 1995 -1996 
    Author : Kiyotoshi SEKIGUCHI, 成宮 周, 祖父江 憲治, 佐邊 壽孝, 目加田 英輔, 宮崎 香, 小澤 政之
     
    (1) Alternative splicing of the fibronectin pre-nRNA at the EDA region was found to regulate the cell-adhesive and integrin alpha5beta1 binding activities of fibronectin. This is the first evidence for the regulation of the integrin-mediated signal transduction by alternative splicing of the extracellular matrix components. (2) The integrin alpha3beta1 binding on laminin-5 was localized in the G2 domain using a series of bacterially produced recombinant fragments derived from different domains of laminin-5. (3) Novel isoforms of paxillin, a crucial adaptor molecule associated with integrins...
  • 文部科学省:科学研究費補助金(重点領域研究)
    Date (from‐to) : 1995 -1995 
    Author : 佐邊 寿孝, 佐邉 寿孝
     
    細胞と細胞外基質間との接着の変化に関連する性質の獲得は癌細胞の悪性度の進行と密接に関連している。本研究は、このような癌の悪性度の進行へ、正常細胞での細胞基質間接着制御機構、及び、その信号伝達機構がどの様に変化し、関与しているかを解明することを目的とする。本年度においては、以下の点を明らかにした。1-1)細胞周期M期でarrestした繊維芽細胞において細胞基質接着斑局在蛋白質paxillinの量的低下を見い出していたが、これが、薬剤処理しないnaturalM期でも起こっていることを確認した。また、同様なことは、各種の繊維芽細胞、内皮細胞、リンパ球細胞でも起こっていることを確認した。1-2)このpaxillinの量的低下は、transcription量の変化ではなく、蛋白質分解である。かつ、paxillinのみが分解し、他の細胞基質斑蛋白質は分解しない。1-3)M期での分解に先立って、paxillinはM期特異的セリンリン酸化を受ける。おそらく、このセリンリン酸化を指標として、ubiquitinationされ分解されることを示唆する結果を得た。かつ、M期からの離脱に伴い、paxillinの速やかな量的回復が確認された。2-1)浮遊系細胞から、従来、繊維芽細胞には知られていないpaxillinのisoformcDNAを複数単離した。genomicDNAの構造解析から、これらは、ex...
  • 文部科学省:科学研究費補助金(特別推進研究)
    Date (from‐to) : 1986 -1990 
    Author : 本庶 佑, 佐邊 寿孝, 野間 隆文, 武田 俊一, 川市 正史, 川上 敏明, 木梨 達雄, 佐辺 寿孝, 松浪 伯禎, 清水 章
     
    骨髄多分化能細胞LyD9をストロ-マ細胞株RP010と同時培養することによって、Bリンパ球への分化誘導系を確立した。分化したLyD9由来B細胞から免疫グロブリンH鎖cDNAを単離し、その構造解析を行った。このH鎖cDNAはLyD9のアロタイプ固有のC領域塩基配列を持ち、V領域は特定のV領域に固まることなく様々なV領域が発現されていることが明らかになった。この系の確立により今後ストロ-マ細胞と骨髄多分化能細胞との間の分化誘導に関与する分子の解析が可能となる。またVDJ recombinationに関与する酵素の一つと考えられるRBPーJk遺伝子の構造解析を行った。マウスにおいては2種類の偽遺伝子と2種類の構造上正しい遺伝子とが存在した。この内の一方はプロセス型遺伝子であり塩基配列上はまったく問題がなかったが、イントロンを完全に欠失したものであった。この遺伝子が発現され機能を持つかどうかについては明らかではない。イントロンを持つ正しい遺伝子は、幾つかの異なるスプライシング様式をもって発現されることが明らかになった。RBPーJkは免疫系のみならず、体の各組織においてmRNAタンパク質共に発現されていることが明らかとなった。RBPーJkと相同の遺伝子をショウジョウバエゲノムから単離しその構造解析を行ったところ、マウスのものとアミノ酸配列で74%以上の相同性があることが明かとなった。こ...
  • 文部科学省:科学研究費補助金(がん特別研究)
    Date (from‐to) : 1988 -1988 
    Author : 奥村 康, 仙道 富士郎, 佐辺 寿孝, 吉田 孝人, 岡田 秀親, 藤原 大美, 白井 俊一, 垣生 園子, 佐藤 昇志, 吉木 敬
     
    宿主の免疫系をはじめとし、腫瘍の発生と抑制に関与する因子が、いまや分子遺伝学レベルで解明されつつある。この班では、特異的、非特異的な生体の腫瘍に対する反応系を、細胞性、液性の両面から解析することを目的に班員を構成し、研究を進めた。吉木、白井班員は、異常な免疫応答を母体とした腫瘍発生モデルを確立し、リンパ腫やリンパ性白血病の発生機序を調べた。そしてレトロウイルスの関与や、ポリクローナルなリンパ球増生と宿主の反応性との相関を明らかにした。藤原班員は、宿主の抗腫瘍反応に関与する細胞群の解析と、腫瘍排除のための細胞間相互作用の解析を行い、CD4T細胞を中心とした反応系を明らかにした。垣生班員は、非特異的なキラー細胞の出現様式とその機能分子の発現調節機序を、分子遺伝学的手法を用い、分化過程を反映するクローンを確立することによって調べた。仙道班員もやはり非特異的な生体反応の主役である多形核白血病の抗腫瘍における役割を判明させた。岡田班員は、CR2レセプターがいかにHTLV-1の感染と発症に関与しているかを調べた。佐辺班員は、リンパ性白血病と強い相関のあるIL-2Rのうちβ受容体をコードする遺伝子単離のための準備を進めた。吉田、佐藤班員らは、宿主に免疫応答を起こさせる腫瘍抗原とその遺伝子の解析を行った。また、奥村班員は、抗腫瘍に関与するいくつかのリンパ球機能分子の遺伝子単離とその発現調節機...
  • 細胞基質間接着の制御機構及びそのシグナル伝達
  • Molecular Mechanism and Signaling of Cell to Substratum Adhesion

Educational Activities

Teaching Experience

  • 研究発表技法Ⅰ
    開講年度 : 2018
    課程区分 : 修士課程
    開講学部 : 医学院
  • Master's Thesis Research in Medical Sciences
    開講年度 : 2018
    課程区分 : 修士課程
    開講学部 : 医学院
    キーワード : シグナル伝達、EMT、細胞内トラフィック、癌、浸潤転移、モデルマウス
  • 研究発表技法Ⅱ
    開講年度 : 2018
    課程区分 : 修士課程
    開講学部 : 医学院
  • Basic Principles of Medicine
    開講年度 : 2018
    課程区分 : 修士課程
    開講学部 : 医学院
    キーワード : シグナル伝達、EMT、細胞内トラフィック、癌、浸潤転移、モデルマウス
  • 研究発表技法Ⅰ
    開講年度 : 2018
    課程区分 : 博士後期課程
    開講学部 : 医学研究科
  • Principles of Medicine
    開講年度 : 2018
    課程区分 : 博士後期課程
    開講学部 : 医学院
    キーワード : シグナル伝達、EMT、細胞内トラフィック、癌、浸潤転移、モデルマウス
  • 研究発表技法Ⅱ
    開講年度 : 2018
    課程区分 : 博士後期課程
    開講学部 : 医学研究科
  • Dissertation Research in Medical Sciences
    開講年度 : 2018
    課程区分 : 博士後期課程
    開講学部 : 医学院
    キーワード : シグナル伝達、EMT、細胞内トラフィック、癌、浸潤転移、モデルマウス
  • 基盤医学研究Ⅱ
    開講年度 : 2018
    課程区分 : 博士後期課程
    開講学部 : 医学研究科
  • BiochemistryⅡ
    開講年度 : 2018
    課程区分 : 学士課程
    開講学部 : 医学部
    キーワード : 分子生物学、細胞生物学
  • 基盤医学研究Ⅰ
    開講年度 : 2018
    課程区分 : 博士後期課程
    開講学部 : 医学研究科
  • 医学総論
    開講年度 : 2018
    課程区分 : 博士後期課程
    開講学部 : 医学研究科
  • 研究発表技法Ⅰ
    開講年度 : 2018
    課程区分 : 博士後期課程
    開講学部 : 医学院
  • 研究発表技法Ⅱ
    開講年度 : 2018
    課程区分 : 博士後期課程
    開講学部 : 医学院
  • 選択実習Ⅰ
    開講年度 : 2018
    課程区分 : 学士課程
    開講学部 : 医学部
  • 診療参加型選択科臨床実習
    開講年度 : 2018
    課程区分 : 学士課程
    開講学部 : 医学部


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