研究者データベース

高林 厚史(タカバヤシ アツシ)
低温科学研究所 生物環境部門
助教

基本情報

所属

  • 低温科学研究所 生物環境部門

職名

  • 助教

学位

  • 博士(生命科学)(京都大学)

ホームページURL

科研費研究者番号

  • 90546417

J-Global ID

職歴

  • 2009年04月 - 現在 北海道大学 低温科学研究所 助教
  • 2007年04月 - 2009年03月 日本学術振興会 特別研究員(PD)
  • 2006年12月 - 2007年03月 京都大学生命科学研究科 教務補佐員
  • 2001年04月 - 2004年03月 日本学術振興会 特別研究員(DC1)

学歴

  •         - 2006年   京都大学   生命科学研究科   統合生命科学専攻
  •         - 2006年   京都大学
  •         - 2001年   京都大学   生命科学研究科   統合生命科学専攻
  •         - 2001年   京都大学
  •         - 1999年   京都大学   農学部   生物機能科学科
  •         - 1999年   京都大学

所属学協会

  • 日本光合成学会   日本植物生理学会   

研究活動情報

論文

  • Makio Yokono, Ikumi Umetani, Atushi Takabayashi, Seiji Akimoto, Ayumi Tanaka
    Photosynthesis Research 1 - 7 2018年04月27日 [査読有り][通常論文]
     
    Recently, we isolated a complex consisting of photosystem II (PSII) and light-harvesting complexes (LHCs) from Nannochloropsis granulata (Umetani et al. Photosynth Res 136:49–61, 2017). This complex contained stress-related protein, Lhcx, as a major component of LHC (Protein ID is Naga_100173g12.1), suggesting that non-photochemical quenching activities may be taking place in the PSII-LHC complex. In this study, we examined the energy transfer dynamics in the isolated LHCs and PSII-LHC complexes, and found substantial quenching capacity. In addition, the LHCs contained low-energy chlorophylls with fluorescence maxima at approximately 710 nm, which may enhance the quenching efficiency in the PSII-LHC. Delayed fluorescence analysis suggested that there was an approximately 50% reduction in energy trapping at the PSII reaction center in the PSII-LHC supercomplex under low-pH condition compared to neutral pH condition. Enhanced quenching may confer a survival advantage in the shallow-water habitat of Nannochloropsis.
  • Ikumi Umetani, Motoshi Kunugi, Makio Yokono, Atsushi Takabayashi, Ayumi Tanaka
    Photosynthesis Research 136 1 49 - 61 2018年04月01日 [査読有り][通常論文]
     
    Diverse light-harvesting complexes (LHCs) have been found in photosynthetic microalgae that originated from secondary endosymbiosis involving primary red algae. However, the associations between LHCs and photosystem I (PSI) and photosystem II (PSII) in these microalgae are not fully understood. Eustigmatophyta is a red algal lineage that appears to have a unique organization in its photosynthetic machinery, consisting of only chlorophyll a and carotenoids that are atypical compared with other closely related groups. In this study, the supramolecular organization of pigment–protein complexes in the eustigmatophyte alga, Nannochloropsis granulata was investigated using Clear Native (CN) PAGE coupled with two-dimensional (2D) SDS-PAGE. Our results showed two slowly migrating green bands that corresponded to PSII supercomplexes, which consisted of reaction centers and LHCs. These green bands were also characterized as PSII complexes by their low temperature fluorescence emission spectra. The protein subunits of the PSII–LHC resolved by 2D CN/SDS-PAGE were analyzed by mass spectrometry, and four different LHC proteins were identified. Phylogenetic analysis of the identified LHC protein sequences revealed that they belonged to four different Lhc groups (1) stress-related Lhcx proteins, (2) fucoxanthin chlorophyll a/c-binding Lhcf proteins, (3) red-shifted Chromera light-harvesting proteins (Red-CLH), and (4) Lhcr proteins, which are commonly found in organisms possessing red algal plastids. This is the first report showing evidence of a pigment–protein supercomplex consisting of PSII and LHCs, and to identify PSII-associated LHC proteins in Nannochloropsis.
  • Yukako Kato, Makio Yokono, Seiji Akimoto, Atsushi Takabayashi, Ayumi Tanaka, Ryouichi Tanaka
    PLANT AND CELL PHYSIOLOGY 58 11 2026 - 2039 2017年11月 [査読有り][通常論文]
     
    Light-harvesting-like (LIL) proteins are a group of proteins that share a consensus amino acid sequence with light-harvesting Chl-binding (LHC) proteins. We hypothesized that they might be involved in photosynthesis-related processes. In order to gain a better understanding of a potential role in photosynthesis-related processes, we examined the most recently identified LIL protein, LIL8/PSB33. Recently, it was suggested that this protein is an auxiliary PSII core protein which is involved in organization of the PSII supercomplex. However, we found that the majority of LIL8/PSB33 was localized in stroma lamellae, where PSI is predominant. Moreover, the PSI antenna sizes measured under visible light were slightly increased in the lil8 mutants which lack LIL8/PSB33 protein. Analysis of fluorescence decay kinetics and fluorescence decay-associated spectra indicated that energy transfer to quenching sites within PSI was partially hampered in these mutants. On the other hand, analysis of the steady-state fluorescence spectra in these mutants indicates that a population of LHCII is energetically disconnected from PSII. Taken together, we suggest that LIL8/PSB33 is involved in the fine-tuning of light harvesting and/or energy transfer around both photosystems.
  • Liping Bai, Takashi Fujishiro, Gangfeng Huang, Urgen Koch, Atsushi Takabayashi, Makio Yokono, Ayumi Tanaka, Tao Xu, Xile Hu, Ulrich Ermler, Seigo Shima
    FARADAY DISCUSSIONS 198 37 - 58 2017年06月 [査読有り][通常論文]
     
    The greenhouse gas and energy carrier methane is produced on Earth mainly by methanogenic archaea. In the hydrogenotrophic methanogenic pathway the reduction of one CO2 to one methane molecule requires four molecules of H-2 containing eight electrons. Four of the electrons from two H-2 are supplied for reduction of an electron carrier F-420, which is catalyzed by F-420-reducing [NiFe]-hydrogenase under nickel-sufficient conditions. The same reaction is catalysed under nickel-limiting conditions by [Fe]-hydrogenase coupled with a reaction catalyzed by F-420-dependent methylene tetrahydromethanopterin dehydrogenase. [Fe]-hydrogenase contains an iron-guanylylpyridinol (FeGP) cofactor for H-2 activation at the active site. FeII of FeGP is coordinated to a pyridinol-nitrogen, an acyl-carbon, two CO and a cysteine-thiolate. We report here on comparative genomic analyses of biosynthetic genes of the FeGP cofactor, which are primarily located in a hmd-co-occurring (hcg) gene cluster. One of the gene products is HcgB which transfers the guanosine monophosphate (GMP) moiety from guanosine triphosphate (GTP) to a pyridinol precursor. Crystal structure analysis of HcgB from Methanococcus maripaludis and its complex with 6-carboxymethyl- 3,5-dimethyl-4-hydroxy-2-pyridinol confirmed the physiological guanylyltransferase reaction. Furthermore, we tested the properties of semi-synthetic [Fe]-hydrogenases using the [Fe]-hydrogenase apoenzyme from several methanogenic archaea and a mimic of the FeGP cofactor. On the basis of the enzymatic reactions involved in the methanogenic pathway, we came up with an idea how the methanogenic pathway could be simplified to develop an artificial methanogenesis system.
  • Ryo Furukawa, Motoshi Kunugi, Kunio Ihara, Atsushi Takabayashi, Ayumi Tanaka
    Genome Announcements 5 10 2017年 [査読有り][通常論文]
     
    Palmophyllum crassum is a little-known green alga, with a unique evolutionary position and distinctive photosynthetic features. Here, we present the complete chloroplast genome sequence of Palmophyllum crassum.
  • Atsushi Takabayashi, Saeka Takabayashi, Kaori Takahashi, Mai Watanabe, Hiroko Uchida, Akio Murakami, Tomomichi Fujita, Masahiko Ikeuchi, Ayumi Tanaka
    PLANT AND CELL PHYSIOLOGY 58 1 2017年01月 [査読有り][通常論文]
     
    The identification of protein complexes is important for the understanding of protein structure and function and the regulation of cellular processes. We used blue-native PAGE and tandemmass spectrometry to identify protein complexes systematically, and built a web database, the protein co-migration database (PCoM-DB, http://pcomdb.lowtem.hokudai.ac.jp/proteins/top), to provide prediction tools for protein complexes. PCoM-DB provides migration profiles for any given protein of interest, and allows users to compare them with migration profiles of other proteins, showing the oligomeric states of proteins and thus identifying potential interaction partners. The initial version of PCoM-DB (launched in January 2013) included protein complex data for Synechocystis whole cells and Arabidopsis thaliana thylakoid membranes. Here we report PCoM-DB version 2.0, which includes new data sets and analytical tools. Additional data are included from whole cells of the pelagic marine picocya-nobacterium Prochlorococcus marinus, the thermophilic cyanobacterium Thermosynechococcus elongatus, the unicellular green alga Chlamydomonas reinhardtii and the bryophyte Physcomitrella patens. The Arabidopsis protein data now include data for intact mitochondria, intact chloroplasts, chloroplast stroma and chloroplast envelopes. The new tools comprise a multiple-protein search form and a heat map viewer for protein migration profiles. Users can compare migration profiles of a protein of interest among different organelles or compare migration profiles among different proteins within the same sample. For Arabidopsis proteins, users can compare migration profiles of a protein of interest with putative homologous proteins from non-Arabidopsis organisms. The updated PCoM-DB will help researchers find novel protein complexes and estimate their evolutionary changes in the green lineage.
  • Noriko Ishikawa, Atsushi Takabayashi, Ko Noguchi, Youshi Tazoe, Hiroshi Yamamoto, Susanne von Caemmerer, Fumihiko Sato, Tsuyoshi Endo
    PLANT AND CELL PHYSIOLOGY 57 10 2020 - 2028 2016年10月 [査読有り][通常論文]
     
    C-4 photosynthesis exhibits efficient CO2 assimilation in ambient air by concentrating CO2 around ribulose 1,5-bisphosphate carboxylase/oxygenase (Rubisco) through a metabolic pathway called the C-4 cycle. It has been suggested that cyclic electron flow (CEF) around PSI mediated by chloroplast NADH dehydrogenase-like complex (NDH), an alternative pathway of photosynthetic electron transport (PET), plays a crucial role in C-4 photosynthesis, although the contribution of NDH-mediated CEF is small in C-3 photosynthesis. Here, we generated NDH-suppressed transformants of a C-4 plant, Flaveria bidentis, and showed that the NDH-suppressed plants grow poorly, especially under low-light conditions. CO2 assimilation rates were consistently decreased in the NDH-suppressed plants under low and medium light intensities. Measurements of non-photochemical quenching (NPQ) of Chl fluorescence, the oxidation state of the reaction center of PSI (P700) and the electrochromic shift (ECS) of pigment absorbance indicated that proton translocation across the thylakoid membrane is impaired in the NDH-suppressed plants. Since proton translocation across the thylakoid membrane induces ATP production, these results suggest that NDH-mediated CEF plays a role in the supply of ATP which is required for C-4 photosynthesis. Such a role is more crucial when the light that is available for photosynthesis is limited and the energy production by PET becomes rate-determining for C-4 photosynthesis. Our results demonstrate that the physiological contribution of NDH-mediated CEF is greater in C-4 photosynthesis than in C-3 photosynthesis, suggesting that the mechanism of PET in C-4 photosynthesis has changed from that in C-3 photosynthesis accompanying the changes in the mechanism of CO2 assimilation.
  • Noriko Ishikawa, Atsushi Takabayashi, Fumihiko Sato, Tsuyoshi Endo
    PHOTOSYNTHESIS RESEARCH 129 3 261 - 277 2016年09月 [査読有り][通常論文]
     
    By concentrating CO2, C-4 photosynthesis can suppress photorespiration and achieve high photosynthetic efficiency, especially under conditions of high light, high temperature, and drought. To concentrate CO2, extra ATP is required, which would also require a change in photosynthetic electron transport in C-4 photosynthesis from that in C-3 photosynthesis. Several analyses have shown that the accumulation of the components of cyclic electron flow (CEF) around photosystem I, which generates the proton gradient across thylakoid membranes (Delta pH) and functions in ATP production without producing NADPH, is increased in various NAD-malic enzyme and NADP-malic enzyme C-4 plants, suggesting that CEF may be enhanced to satisfy the increased need for ATP in C-4 photosynthesis. However, in C-4 plants, the accumulation patterns of the components of two partially redundant pathways of CEF, NAD(P)H dehydrogenase-like complex and PROTON GRADIENT REGULATION5-PGR5-like1 complex, are not identical, suggesting that these pathways may play different roles in C-4 photosynthesis. Accompanying the increase in the amount of NDH, the expression of some genes which encode proteins involved in the assembly of NDH is also increased at the mRNA level in various C-4 plants, suggesting that this increase is needed to increase the accumulation of NDH. To better understand the relation between CEF and C-4 photosynthesis, a reverse genetic approach to generate C-4 transformants with respect to CEF will be necessary.
  • Atsushi Takabayashi, Akihiro Niwata, Ayumi Tanaka
    SCIENTIFIC REPORTS 6 2016年07月 [査読有り][通常論文]
     
    Because it plays an essential role in nitrogen (N) assimilation and photorespiration, the glutamine synthetase (GS)/glutamate synthase (GOGAT) system is widely accepted as occupying a central position in leaf N metabolism. However, the regulation of GOGAT at the post-transcriptional level is poorly understood. Here, we show that ACR11, an ACT (acronym for aspartate kinase, chorismate mutase, and TyrA) domain-containing family protein, interacts with Glu1-encoded ferredoxin (Fd)-GOGAT in Arabidopsis chloroplasts. In addition, Arabidopsis acr11 mutants have lost the capability to control Fd-GOGAT levels in response to light/dark diurnal cycles, nitrogen inputs, and changes in photorespiratory activity. Considering that ACR11 has putative glutamine-binding domains, our results indicate that ACR11 is necessary for post-transcriptional control of leaf Glu1-encoded Fd-GOGAT. This regulation takes place through direct interaction of ACR11 and Fd-GOGAT, possibly in an allosteric manner.
  • Motoshi Kunugi, Soichirou Satoh, Kunio Ihara, Kensuke Shibata, Yukimasa Yamagishi, Kazuhiro Kogame, Junichi Obokata, Atsushi Takabayashi, Ayumi Tanaka
    PLANT AND CELL PHYSIOLOGY 57 6 1231 - 1243 2016年06月 [査読有り][通常論文]
     
    Photosynthetic organisms have various pigments enabling them to adapt to various light environments. Green plants are divided into two groups: streptophytes and chlorophytes. Streptophytes include some freshwater green algae and land plants, while chlorophytes comprise the other freshwater green algae and seawater green algae. The environmental conditions driving the divergence of green plants into these two groups and the changes in photosynthetic properties accompanying their evolution remain unknown. Here, we separated the core antennae of PSI and the peripheral antennae [light-harvesting complexes (LHCs)] in green plants by green-native gel electrophoresis and determined their pigment compositions. Freshwater green algae and land plants have high Chl a/b ratios, with most Chl b existing in LHCs. In contrast, seawater green algae have low Chl a/b ratios. In addition, Chl b exists not only in LHCs but also in PSI core antennae in these organisms, a situation beneficial for survival in deep seawater, where blue-green light is the dominant light source. Finally, low-energy Chl (red Chl) of PSI was detected in freshwater green algae and land plants, but not in seawater green algae. We thus conclude that the different level of Chl b accumulation in core antennae and differences in PSI red Chl between freshwater and seawater green algae are evolutionary adaptations of these algae to their habitats, especially to high-or low-light environments.
  • M. Yokono, A. Takabayashi, S. Akimoto, A. Tanaka
    NATURE COMMUNICATIONS 6 2015年03月 [査読有り][通常論文]
     
    Throughout the history of oxygen evolution, two types of photosystem reaction centres (PSI and PSII) have worked in a coordinated manner. The oxygen evolving centre is an integral part of PSII, and extracts an electron from water. PSI accepts the electron, and accumulates reducing power. Traditionally, PSI and PSII are thought to be spatially dispersed. Here, we show that about half of PSIIs are physically connected to PSIs in Arabidopsis thaliana. In the PSI-PSII complex, excitation energy is transferred efficiently between the two closely interacting reaction centres. PSII diverts excitation energy to PSI when PSII becomes closed-state in the PSI-PSII complex. The formation of PSI-PSII complexes is regulated by light conditions. Quenching of excess energy by PSI might be one of the physiological functions of PSI-PSII complexes.
  • Shugo Maekawa, Atsushi Takabayashi, Thais Huarancca Reyes, Hiroko Yamamoto, Ayumi Tanaka, Takeo Sato, Junji Yamaguchi
    PLOS ONE 10 2 2015年02月 [査読有り][通常論文]
     
    Arabidopsis ubiquitin ligases ATL31 and homologue ATL6 control the carbon/nitrogen nutrient and pathogen responses. A mutant with the loss-of-function of both atl31 and atl6 developed light intensity-dependent pale-green true leaves, whereas the single knockoutmutants did not. Plastid ultrastructure and Blue Native-PAGE analyses revealed that pale-green leaves contain abnormal plastid structure with highly reduced levels of thylakoid proteins. In contrast, the pale-green leaves of the atl31/atl6 mutant showed normal Fv/Fm. In the pale-green leaves of the atl31/atl6, the expression of HEMA1, which encodes the key enzyme for 5-aminolevulinic acid synthesis, the rate-limiting step in chlorophyll biosynthesis, was markedly down-regulated. The expression of key transcription factor GLK1, which directly promotes HEMA1 transcription, was also significantly decreased in atl31/atl6 mutant. Finally, application of 5-aminolevulinic acid to the atl31/atl6 mutants resulted in recovery to a green phenotype. Taken together, these findings indicate that the 5-aminolevulinic acid biosynthesis step was inhibited through the down-regulation of chlorophyll biosynthesis-related genes in the pale-green leaves of atl31/atl6 mutant.
  • Kaori Takahashi, Atsushi Takabayashi, Ayumi Tanaka, Ryouichi Tanaka
    JOURNAL OF BIOLOGICAL CHEMISTRY 289 2 987 - 999 2014年01月 [査読有り][通常論文]
     
    The light-harvesting complex (LHC) constitutes the major light-harvesting antenna of photosynthetic eukaryotes. LHC contains a characteristic sequence motif, termed LHC motif, consisting of 25-30 mostly hydrophobic amino acids. This motif is shared by a number of transmembrane proteins from oxygenic photoautotrophs that are termed light-harvesting-like (LIL) proteins. To gain insights into the functions of LIL proteins and their LHC motifs, we functionally characterized a plant LIL protein, LIL3. This protein has been shown previously to stabilize geranylgeranyl reductase (GGR), a key enzyme in phytol biosynthesis. It is hypothesized that LIL3 functions to anchor GGR to membranes. First, we conjugated the transmembrane domain of LIL3 or that of ascorbate peroxidase to GGR and expressed these chimeric proteins in an Arabidopsis mutant lacking LIL3 protein. As a result, the transgenic plants restored phytol-synthesizing activity. These results indicate that GGR is active as long as it is anchored to membranes, even in the absence of LIL3. Subsequently, we addressed the question why the LHC motif is conserved in the LIL3 sequences. We modified the transmembrane domain of LIL3, which contains the LHC motif, by substituting its conserved amino acids (Glu-171, Asn-174, and Asp-189) with alanine. As a result, the Arabidopsis transgenic plants partly recovered the phytol-biosynthesizing activity. However, in these transgenic plants, the LIL3-GGR complexes were partially dissociated. Collectively, these results indicate that the LHC motif of LIL3 is involved in the complex formation of LIL3 and GGR, which might contribute to the GGR reaction.
  • Motoshi Kunugi, Atsushi Takabayashi, Ayumi Tanaka
    JOURNAL OF BIOLOGICAL CHEMISTRY 288 27 19330 - 19341 2013年07月 [査読有り][通常論文]
     
    Chlorophyll b is found in photosynthetic prokaryotes and primary and secondary endosymbionts, although their light-harvesting systems are quite different. Chlorophyll b is synthesized from chlorophyll a by chlorophyllide a oxygenase (CAO), which is a Rieske-mononuclear iron oxygenase. Comparison of the amino acid sequences of CAO among photosynthetic organisms elucidated changes in the domain structures of CAO during evolution. However, the evolutionary relationship between the light-harvesting system and the domain structure of CAO remains unclear. To elucidate this relationship, we investigated the CAO structure and the pigment composition of chlorophyll-protein complexes in the prasinophyte Micromonas. The Micromonas CAO is composed of two genes, MpCAO1 and MpCAO2, that possess Rieske and mononuclear iron-binding motifs, respectively. Only when both genes were introduced into the chlorophyll b-less Arabidopsis mutant (ch1-1) was chlorophyll b accumulated, indicating that cooperation between the two subunits is required to synthesize chlorophyll b. Although Micromonas has a characteristic light-harvesting system in which chlorophyll b is incorporated into the core antennas of reaction centers, chlorophyll b was also incorporated into the core antennas of reaction centers of the Arabidopsis transformants that contained the two Micromonas CAO proteins. Based on these results, we discuss the evolutionary relationship between the structures of CAO and light-harvesting systems.
  • Atsushi Takabayashi, Ryosuke Kadoya, Masayoshi Kuwano, Katsunori Kurihara, Hisashi Ito, Ryouichi Tanaka, Ayumi Tanaka
    SPRINGERPLUS 2 2013年 [査読有り][通常論文]
     
    Protein-protein interactions are critical for most cellular processes; however, many remain to be identified. Here, to comprehensively identify protein complexes in photosynthetic organisms, we applied the recently developed approach of blue native PAGE (BN-PAGE) coupled with LC-MS/MS to the thylakoid proteins of Arabidopsis thaliana and the whole cell proteins of whole cell proteins of Synechocystis sp. PCC 6803. We identified 245 proteins from the purified Arabidopsis thylakoid membranes and 1,458 proteins from the whole cells of Synechocystis using the method. Next, we generated protein migration profiles that were assessed by plotting the label-free estimations of protein abundances versus migration distance in BN-PAGE. Comparisons between the migration profiles of the major photosynthetic complexes and their band patterns showed that the protein migration profiles were well correlated. Thus, the protein migration profiles allowed us to estimate the molecular size of each protein complex and to identify co-migrated proteins with the proteins of interest by determining the protein pairs that contained peaks in the same gel slice. Finally, we built the protein co-migration database for photosynthetic organisms (PCoM-DB: http://pcomdb.lowtem.hokudai.ac.jp/proteins/top) to make our data publicly accessible online, which stores the analyzed data with a user-friendly interface to compare the migration profiles of proteins of interest. It helps users to find unidentified protein complexes in Arabidopsis thylakoids and Synechocystis cells. The accumulation of the data from the BN-PAGE coupled with LC-MS/MS should reveal unidentified protein complexes and should aid in understanding the adaptation and the evolution of photosynthetic organisms.
  • Atsushi Takabayashi, Katsunori Kurihara, Masayoshi Kuwano, Yasuhiro Kasahara, Ryouichi Tanaka, Ayumi Tanaka
    PLANT AND CELL PHYSIOLOGY 52 12 2103 - 2114 2011年12月 [査読有り][通常論文]
     
    The reversible associations between the light-harvesting complexes (LHCs) and the core complexes of PSI and PSII are essential for the photoacclimation mechanisms in higher plants. Two types of Chls, Chl a and Chl b, both function in light harvesting and are required for the biogenesis of the photosystems. Chl b-less plants have been studied to determine the function of the LHCs because the Chl b deficiency has severe effects specific to the LHCs. Previous studies have shown that the amounts of the LHCs, especially the LHCII trimer, were decreased in the mutants; however, it is still unclear whether Chl b is required for the assembly of the LHCs and for the association of the LHCs with PSI and PSII. Here, to reveal the function of Chl b in the LHCs, we investigated the oligomeric states of the LHCs, PSI and PSII in the Arabidopsis Chl b-less mutant. A two-dimensional blue native-PAGE/SDS-PAGE demonstrated that the PSI-LHCI supercomplex was fully assembled in the absence of Chl b, whereas the trimeric LHCII and PSII-LHCII supercomplexes were not detected. The PSI-NAD(P)H dehydrogenase (NDH) supercomplexes were also assembled in the mutant. Furthermore, we detected two forms of monomeric LHC proteins. The faster migrating forms, which were detected primarily in the mutant, were probably apo-LHC proteins, whereas the slower migrating forms were probably the LHC proteins that contained Chl a. These findings increase our understanding of the Chl b function in the assembly of LHCs and the association of the LHCs with PSI, PSII and NDH.
  • Satoshi Ishida, Ken-Ichi Morita, Masahiro Kishine, Atsushi Takabayashi, Reiko Murakami, Satomi Takeda, Ko Shimamoto, Fumihiko Sato, Tsuyoshi Endo
    PLANT AND CELL PHYSIOLOGY 52 10 1822 - 1831 2011年10月 [査読有り][通常論文]
     
    The thermal dissipation (TD) of absorbed light energy in PSII is considered to be an important photoprotection process in photosynthesis. A major portion of TD has been visualized through the analysis of Chl fluorescence as energy quenching (qE) which depends on the presence of the PsbS subunit. Although the physiological importance of qE-associated TD (qE-TD) has been widely accepted, it is not yet clear how much of the absorbed light energy is dissipated through a qE-associated mechanism. In this study, the fates of absorbed light energy in PSII with regard to different TD processes, including qE-TD, were quantitatively estimated by the typical energy allocation models using transgenic rice in which psbS genes were silenced by RNA interference (RNAi). The silencing of psbS genes resulted in a decrease in the light-inducible portion of TD, whereas the allocation of energy to electron transport did not change over a wide range of light intensities. The allocation models indicate that the energy allocated to qE-TD under saturating light is 30-50%. We also showed that a large portion of absorbed light energy is thermally dissipated in manners that are independent of qE. The nature of such dissipations is discussed.
  • Miki Meguro, Hisashi Ito, Atsushi Takabayashi, Ryouichi Tanaka, Ayumi Tanaka
    PLANT CELL 23 9 3442 - 3453 2011年09月 [査読有り][通常論文]
     
    The interconversion of chlorophyll a and chlorophyll b, referred to as the chlorophyll cycle, plays a crucial role in the processes of greening, acclimation to light intensity, and senescence. The chlorophyll cycle consists of three reactions: the conversions of chlorophyll a to chlorophyll b by chlorophyllide a oxygenase, chlorophyll b to 7-hydroxymethyl chlorophyll a by chlorophyll b reductase, and 7-hydroxymethyl chlorophyll a to chlorophyll a by 7-hydroxymethyl chlorophyll a reductase. We identified 7-hydroxymethyl chlorophyll a reductase, which is the last remaining unidentified enzyme of the chlorophyll cycle, from Arabidopsis thaliana by genetic and biochemical methods. Recombinant 7-hydroxymethyl chlorophyll a reductase converted 7-hydroxymethyl chlorophyll a to chlorophyll a using ferredoxin. Both sequence and biochemical analyses showed that 7-hydroxymethyl chlorophyll a reductase contains flavin adenine dinucleotide and an iron-sulfur center. In addition, a phylogenetic analysis elucidated the evolution of 7-hydroxymethyl chlorophyll a reductase from divinyl chlorophyllide vinyl reductase. A mutant lacking 7-hydroxymethyl chlorophyll a reductase was found to accumulate 7-hydroxymethyl chlorophyll a and pheophorbide a. Furthermore, this accumulation of pheophorbide a in the mutant was rescued by the inactivation of the chlorophyll b reductase gene. The downregulation of pheophorbide a oxygenase activity is discussed in relation to 7-hydroxymethyl chlorophyll a accumulation.
  • Ryouichi Tanaka, Maxi Rothbart, Seiko Oka, Atsushi Takabayashi, Kaori Takahashi, Masaru Shibata, Fumiyoshi Myouga, Reiko Motohashi, Kazuo Shinozaki, Bernhard Grimm, Ayumi Tanaka
    PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA 107 38 16721 - 16725 2010年09月 [査読有り][通常論文]
     
    The light-harvesting chlorophyll-binding (LHC) proteins are major constituents of eukaryotic photosynthetic machinery. In plants, six different groups of proteins, LHC-like proteins, share a conserved motif with LHC. Although the evolution of LHC and LHC-like proteins is proposed to be a key for the diversification of modern photosynthetic eukaryotes, our knowledge of the evolution and functions of LHC-like proteins is still limited. In this study, we aimed to understand specifically the function of one type of LHC-like proteins, LIL3 proteins, by analyzing Arabidopsis mutants lacking them. The Arabidopsis genome contains two gene copies for LIL3, LIL3:1 and LIL3:2. In the lil3:1/lil3:2 double mutant, the majority of chlorophyll molecules are conjugated with an unsaturated geranylgeraniol side chain. This mutant is also deficient in a-tocopherol. These results indicate that reduction of both the geranylgeraniol side chain of chlorophyll and geranylgeranyl pyrophosphate, which is also an essential intermediate of tocopherol biosynthesis, is compromised in the lil3 mutants. We found that the content of geranylgeranyl reductase responsible for these reactions was severely reduced in the lil3 double mutant, whereas the mRNA level for this enzyme was not significantly changed. We demonstrated an interaction of geranylgeranyl reductase with both LIL3 isoforms by using a split ubiquitin assay, bimolecular fluorescence complementation, and combined blue-native and SDS polyacrylamide gel electrophoresis. We propose that LIL3 is functionally involved in chlorophyll and tocopherol biosynthesis by stabilizing geranylgeranyl reductase.
  • Shinya Yabuta, Kentaro Ifuku, Atsushi Takabayashi, Seiko Ishihara, Kunio Ido, Noriko Ishikawa, Tsuyoshi Endo, Fumihiko Sato
    PLANT AND CELL PHYSIOLOGY 51 6 866 - 876 2010年06月 [査読有り][通常論文]
     
    Arabidopsis has three PsbQ-like (PQL) proteins in addition to the PsbQ subunit of the oxygen-evolving complex of PSII. Recent bioinformatic and proteomic studies suggested that the two PQL proteins, PQL1 (At1g14150) and PQL2 (At3g01440), might function in the chloroplast NAD(P)H dehydrogenase (NDH) complex; however, their molecular function has not been characterized. In this study, we examined the function of the chloroplast NDH in the Arabidopsis pql1 and pql2 mutants. Post-illumination increases in Chl fluorescence, which are caused by an NDH-dependent cyclic electron flow, were absent in both mutants, indicating that PQL1 and PQL2 are required for NDH activity. In the thylakoid membranes of wild-type plants, PQL1 and PQL2 were tightly associated with the NDH PSI supercomplex and protected from protease treatments, while unassembled PQLs were not stably accumulated in mutants lacking known NDH subunits. Subunit stability of the NDH complex was affected differently in the thylakoid membranes of the pql1 and pql2 mutants. These data indicate that PQL1 and PQL2 are novel NDH subunits and differ in their functional roles and in their binding sites in the NDH complex. Furthermore, functional analysis on PQL3 (At2g01918) using the pql3 mutant suggests that PQL3 is also required for NDH activity. Proteins homologous to each PQL protein are found in various plant species, but not in cyanobacteria, algae, mosses or ferns. These results suggest that seed plants that have NDH activity in chloroplasts specifically developed three PQL proteins for the function of the chloroplast NDH complex.
  • Risa Mutoh, Hiroyuki Mino, Reiko Murakami, Tatsuya Uzumaki, Atsushi Takabayashi, Kentaro Ishii, Masahiro Ishiura
    GENES TO CELLS 15 3 269 - 280 2010年03月 [査読有り][通常論文]
     
    In cyanobacteria, three clock proteins, KaiA, KaiB and KaiC, play essential roles in generating circadian oscillations. The interactions of these proteins change during the circadian cycle. Here, we demonstrated direct interaction between KaiA and KaiB using electron spin resonance spectroscopy. We prepared cystein (Cys)-substituted mutants of Thermosynechococcus elongatus KaiB, labeled specifically their Cys residues with spin labels and measured the ESR spectra of the labeled KaiB. We found that KaiB labeled at the 64th residue showed spectral changes in the presence of KaiA, but not in the presence of KaiC or bovine serum albumin as a negative control. KaiB labeled at the 101st residue showed no such spectral changes even in the presence of KaiA. The results suggest that KaiB interacts with KaiA in the vicinity of the 64th residue of KaiB. Further analysis demonstrated that the C-terminal clock-oscillator domain of KaiA is responsible for this interaction.
  • Satoshi Ishida, Atsushi Takabayashi, Noriko Ishikawa, Yasushi Hano, Tsuyoshi Endo, Fumihiko Sato
    PLANT AND CELL PHYSIOLOGY 50 2 383 - 393 2009年02月 [査読有り][通常論文]
     
    The chloroplast NAD(P)H dehydrogenase (NDH) complex, which reduces plastoquinones in thylakoid membranes, is involved in PSI cyclic electron flow and chlororespiration. In addition to land plants, the NDH complex is conserved in cyanobacteria. In this study, we identified a novel NDH-related gene of Arabidopsis, NDH-dependent cyclic electron flow 5 (NDF5, At1g55370). Post-illumination increases in chlorophyll fluorescence were absent in ndf5 mutant plants, which indicated that NDF5 is essential for NDH activity. Sequence analysis did not reveal any known functional motifs in NDF5, but there was some homology in amino acid sequence between NDF5 and NDF2, a known NDH subunit. NDF5 and NDF2 homologs were present in higher plants, but not cyanobacteria. A single homolog, which had similarity to both NDF5 and NDF2, was identified in the moss Physcomitrella patens. Immunoblot analysis showed that NDF5 localizes to membrane fractions of chloroplasts. The stability of NdhH, a subunit of the NDH complex, as well as NDF5 and NDF2, was decreased in ndf5, ndf2 and double ndf2ndf5 mutants, resulting in a loss of NDH activity in these mutants. These results indicated that both NDF5 and NDF2 have essential functions in the stabilization of the NDH complex. We propose that NDF5 and NDF2 were acquired by land plants during evolution, and that in higher plants both NDF5 and NDF2are critical to regulate NDH activity and each others protein stability, as well as the stability of additional NDH subunits.
  • Atsushi Takabayashi, Noriko Ishikawa, Takeshi Obayashi, Satoshi Ishida, Junichi Obokata, Tsuyoshi Endo, Fumihiko Sato
    PLANT JOURNAL 57 2 207 - 219 2009年01月 [査読有り][通常論文]
     
    Chloroplastic NAD(P)H dehydrogenase (NDH) plays a role in cyclic electron flow around photosystem I to produce ATP, especially in adaptation to environmental changes. Although the NDH complex contains 11 subunits that are homologous to NADH:ubiquinone oxidoreductase (complex I; EC 1.6.5.3), recent genetic and biological studies have indicated that NDH also comprises unique subunits. We describe here an in silico approach based on co-expression analysis and phylogenetic profiling that was used to identify 65 genes as potential candidates for NDH subunits. Characterization of 21 Arabidopsis T-DNA insertion mutants among these ndh gene candidates indicated that three novel ndf (NDH-dependent cyclic electron flow) mutants (ndf1, ndf2 and ndf4) had impaired NDH activity as determined by measurement of chlorophyll fluorescence. The amount of NdhH subunit was greatly decreased in these mutants, suggesting that the loss of NDH activity was caused by a defect in accumulation of the NDH complex. In addition, NDF1, NDF2 and NDF4 proteins co-migrated with the NdhH subunit, as shown by blue native electrophoresis. These results strongly suggest that NDF proteins are novel subunits of the NDH complex. Further analysis revealed that the NDF1 and NDF2 proteins were unstable in the mutants lacking hydrophobic subunits of the NDH complex, but were stable in mutants lacking the hydrophilic subunits, suggesting that NDF1 and NDF2 interact with a hydrophobic sub-complex. NDF4 protein was predicted to possess a redox-active iron-sulfur cluster domain that may be involved in the electron transfer.
  • Noriko Ishikawa, Atsushi Takabayashi, Satoshi Ishida, Yasushi Hano, Tsuyoshi Endo, Fumihiko Sato
    PLANT AND CELL PHYSIOLOGY 49 7 1066 - 1073 2008年07月 [査読有り][通常論文]
     
    NAD(P)H dehydrogenase (NDH) is a homolog of respiratory complex I and mediates one of the two pathways of cyclic electron flow around PSI (CEF I). Although 15 ndh subunits have been identified in the chloroplastic and nuclear genomes of higher plants, no electron accepter subunits have been identified to date. To identify the missing chloroplastic NDH subunits, we undertook an in silico approach based on co-expression analysis. In this report, we characterized the novel gene NDF6 (NDH-dependent flow 6; At1g18730) which encodes a protein that is essential for NDH activity. NDF6 has one transmembrane domain and is localized in the thylakoid membrane fraction. Homologous proteins of NDF6 were identified in the genomes of terrestrial plants; however, no homologs have been found in cyanobacteria, which are thought to be the origin of chloroplasts and have a minimal NDH complex unit. NDF6 is unstable in ndhB-impaired or disrupted mutants of higher plants in which the chloroplastic NDH complex is thought to be degraded. These results suggest that NDF6 is a novel subunit of chloroplastic NDH that was added to terrestrial plants during evolution.
  • Seiko Ishihara, Atsushi Takabayashi, Kunio Ido, Tsuyoshi Endo, Kentaro Ifuku, Fumihiko Sato
    PLANT PHYSIOLOGY 145 3 668 - 679 2007年11月 [査読有り][通常論文]
     
    PsbP, an extrinsic subunit of photosystem II ( PSII), is a nuclear- encoded protein that optimizes the water- splitting reaction in vivo. In addition to PsbP, higher plants have two nuclear- encoded genes for PsbP homologs ( PsbP- like proteins [ PPLs]) that show significant sequence similarity to a cyanobacterial PsbP homolog ( cyanoP); however, the function of PPLs in higher plants has not yet been elucidated. In this study, we characterized Arabidopsis ( Arabidopsis thaliana) mutants lacking either of two PPLs, PPL1 and PPL2. Phylogenetic analysis suggests that PPL1 would be an ortholog of cyanoP, and PPL2 and PsbP may have a paralogous relationship with PPL1. Analysis on mRNA expression profiles showed that PPL1 expressed under stress conditions and PPL2 coexpressed with the subunits of chloroplast NAD( P) H dehydrogenase ( NDH) complex. Consistent with these suggestions, PSII activity in a ppl1 mutant was more sensitive to high- intensity light than wild type, and the recovery of photoinhibited PSII activity was delayed in ppl1 plants. Therefore, PPL1 is required for efficient repair of photodamaged PSII. Furthermore, the stoichiometric level and activity of the chloroplast NDH complex in thylakoids were severely decreased in a ppl2 mutant, demonstrating that PPL2 is a novel thylakoid lumenal factor required for accumulation of the chloroplast NDH complex. These results suggest that during endosymbiosis and subsequent gene transfer to the host nucleus, cyanoP from ancient cyanobacteria evolved into PPL1, PPL2, and PsbP, and each of them has a distinct role in photosynthetic electron transfer in Arabidopsis.
  • P Wang, W Duan, A Takabayashi, T Endo, T Shikanai, JY Ye, HL Mi
    PLANT PHYSIOLOGY 141 2 465 - 474 2006年06月 [査読有り][通常論文]
     
    In this study, the function of the NAD( P) H dehydrogenase ( NDH)-dependent pathway in suppressing the accumulation of reactive oxygen species in chloroplasts was investigated. Hydrogen peroxide accumulated in the leaves of tobacco ( Nicotiana tabacum) defective in ndhC-ndhK-ndhJ ( Delta ndhCKJ) at 42 degrees C and 4 degrees C, and in that of wild-type leaves at 4 degrees C. The maximum quantum efficiency of PSII decreased to a similar extent in both strains at 42 degrees C, while it decreased more evidently in DndhCKJ at 4 degrees C. The parameters linked to CO2 assimilation, such as the photochemical efficiency of PSII, the decrease of nonphotochemical quenching following the initial rise, and the photosynthetic O-2 evolution, were inhibited more significantly in DndhCKJ than in wild type at 42 degrees C and were seriously inhibited in both strains at 4 degrees C. While cyclic electron flow around PSI mediated by NDH was remarkably enhanced at 42 degrees C and suppressed at 4 degrees C. The proton gradient across the thylakoid membranes and light-dependent ATP synthesis were higher in wild type than in DndhCKJ at either 25 degrees C or 42 degrees C, but were barely formed at 4 degrees C. Based on these results, we suggest that cyclic photophosphorylation via the NDH pathway might play an important role in regulation of CO2 assimilation under heat-stressed condition but is less important under chilling-stressed condition, thus optimizing the photosynthetic electron transport and reducing the generation of reactive oxygen species.
  • A Takabayashi, M Kishine, K Asada, T Endo, F Sato
    PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA 102 46 16898 - 16903 2005年11月 [査読有り][通常論文]
     
    Whereas linear electron flow (LEF) in photosynthesis produces both ATP and NADPH, the cyclic electron flow (CEF) around photosystem I has been shown to produce only ATP. Two alternative routes have been shown for CEF; NAD(P)H dehydrogenase (NDH) and ferredoxin:plastoquinone oxicloreductase (FQR)-dependent flows, but their physiological relevance has not been elucidated in detail. Meanwhile, because C-4 photosynthesis requires more ATP than does C-3 photosynthesis to concentrate CO2, it has not been clear how the extra ATP is produced. In this study, to elucidate whether CEF contributes to the additional ATP needed in C-4 photosynthesis, we estimated the amounts of PGR5, which participates in FQR-dependent flow, and NDH-H, a subunit of NDH, in four C-4 species. Although the expression profiles of PGR5 did not correlate well with the additional ATP requirement, NDH was greatly expressed in mesophyll cells in the NAD-malic enzyme (ME) species, and in bundle-sheath cells in NADP-ME species, where there is a strong need for ATP in the respective cells. Our results indicate that CEF via NDH plays a central role in driving the CO2-concentrating mechanism in C-4 photosynthesis.
  • R Murakami, K Ifuku, A Takabayashi, T Shikanai, T Endo, F Sato
    FEBS JOURNAL 272 9 2165 - 2175 2005年05月 [査読有り][通常論文]
     
    PsbO protein is an extrinsic subunit of photosystem II (PSII) and has been proposed to play a central role in stabilization of the catalytic manganese cluster. Arabidopsis thaliana has two psbO genes that express two PsbO proteins; PsbO1 and PsbO2. We reported previously that a mutant plant that lacked PsbO1 (psbo1) showed considerable growth retardation despite the presence of PsbO2 [Murakami, R., Ifuku, K., Takabayashi, A., Shikanai, T., Endo, T., and Sato, F. (2002) FEBS Lett523, 138-142]. In the present study, we characterized the functional differences between PsbO1 and PsbO2. We found that PsbO1 is the major isoform in the wild-type, and the amount of PsbO2 in psbo1 was significantly less than the total amount of PsbO in the wild-type. The amount of PsbO as well as the efficiency of PSII in psbo1 increased as the plants grew; howeVER, it neVER reached the total PsbO level observed in the wild-type, suggesting that the poor activity of PSII in psbo1 was caused by a shortage of PsbO. In addition, an in vitro reconstitution experiment using recombinant PsbOs and urea-washed PSII particles showed that oxygen evolution was better recoVERed by PsbO1 than by PsbO2. Further analysis using chimeric and mutated PsbOs suggested that the amino acid changes Val186 -> Ser, Leu246 -> Ile, and Val204 -> Ile could explain the functional difference between the two PsbOs. Therefore we concluded that both the lower expression level and the inferior functionality of PsbO2 are responsible for the phenotype observed in psbo1.
  • M Kishine, A Takabayashi, Y Munekage, T Shikanai, T Endo, F Sato
    PLANT MOLECULAR BIOLOGY 55 4 595 - 606 2004年07月 [査読有り][通常論文]
     
    An Arabidopsis mutant rnr1, which has a defect in the basic genetic system in chloroplasts, was isolated using the screening of the high chlorophyll fluorescence phenotype. Whereas chlorophyll fluorescence and immunoblot studies showed the mutant had reduced activities of photosystems I and II, molecular characterization of the mutant suggested that a T-DNA insertion impaired the expression of a gene encoding a RNase R family member with a targeting signal to chloroplasts. Since RNase R family members have a 3'-5'exoribonuclease activity, we examined the RNA profile in chloroplasts. In rnr1 the intercistronic cleavage between 23S and 4.5S rRNA was impaired, and a significant reduction in rRNA in chloroplasts was found, suggesting that RNR1 functions in the maturation of chloroplast rRNA. The present results suggest that defects in the genetic system in chloroplasts cause high chlorophyll fluorescence, pale green leaf, and marked reduction in the growth rate, whereas the levels of some chloroplast RNA were higher in rnr1 than in the wild-type.
  • A Takabayashi, T Endo, T Shikanai, F Sato
    BIOSCIENCE BIOTECHNOLOGY AND BIOCHEMISTRY 66 10 2107 - 2111 2002年10月 [査読有り][通常論文]
     
    We reported previously that an ndhB gene disruptant, DeltandhB, had the same phenotype as wild-type tobacco plants under normal growth conditions. Two other groups have reported conflicting phenotypes with each other for ndhCKJ operon disruptants. Here, we generated two transformants in which the ndhCKJ operon was disrupted, and found that new transformants had the same phenotype as DeltandhB. After illumination with visible light, all ndh disruptants had higher levels of steady-state fluorescence than wild-type controls when measured under weak light, suggesting that reduction of the plastoquinone pool in ndh disruptants was greater than that in wild-type controls. The weak light itself could not reduce the plastoquinone much, so the reduction in the plastoquinone in the mutant was due to electron donation from stromal reductants generated during illumination with the strong light. These results supported the hypothesis that NAD(P)H dehydrogenase prevents overreduction in chloroplasts and suggested that chlororespiratory oxidase did not function under low light or in the dark.
  • R Murakami, K Ifuku, A Takabayashi, T Shikanai, T Endo, F Sato
    FEBS LETTERS 523 1-3 138 - 142 2002年07月 [査読有り][通常論文]
     
    A 33-kDa protein component of the oxygen-evolving complex in photosystem 11 is essential for photosynthesis, and it has been believed that mutants with deletion of this 33-kDa protein are not found in higher plants. We report here the first isolation of an Arabidopsis thaliana mutant with a defect in one of the genes for the 33-kDa proteins, psbO, and an intact gene (psbO2). This mutant showed considerable growth retardation, suggesting that there is a functional difference between psbO and psbO2. (C) 2002 Federation of European Biochemical Societies. Published by Elsevier Science B.V. All rights reserved.
  • T Endo, T Shikanai, A Takabayashi, K Asada, F Sato
    FEBS LETTERS 457 1 5 - 8 1999年08月 [査読有り][通常論文]
     
    After a brief exposure to supra-saturating light, leaves of a tobacco transformant, in which chloroplastic NAD(P)H dehydrogenase (NDH) Tvas defective, showed more severe photoinhibition than the wild-type, when judged by the parameter of chlorophyll fluorescence Fv/Fm, Repeated application of supra-saturating light eventually resulted in chlorosis in the NDH-defective mutant, while the mild-type sustained less photodamage and was able to recover from it. The mechanism of the phenomena is discussed with respect to the potential role of NDH in photosynthesis. (C) 1999 Federation of European Biochemical Societies.

その他活動・業績

  • 光化学系の光環境適応とその進化的な制約 -緑藻の光化学系をモデルとして-
    高林 厚史 光合成研究 2017年12月 [査読有り][招待有り]
  • ブルーネイティブ電気泳動を用いた光合成生物タンパク質複合体の網羅的解析
    高林 厚史, 田中 歩 電気泳動 61 (2) 111 -114 2017年11月 [査読有り][招待有り]
  • ゲノム世代のタンパク質複合体解析
    光合成研究 60 2010年 [査読無し][通常論文]
  • 地球環境と光合成の共進化
    低温科学 21 2010年 [査読無し][通常論文]
  • 発現プロファイルの利用
    低温科学 663 2009年 [査読無し][通常論文]

共同研究・競争的資金等の研究課題

  • 葉の窒素代謝ネットワークの転写後制御の解明とその応用
    文部科学省:基盤研究(C)
    研究期間 : 2017年 -2019年 
    代表者 : 高林 厚史
  • 植物の新規窒素代謝制御機構の解明とその応用
    文部科学省:若手研究(B)
    研究期間 : 2014年 -2016年 
    代表者 : 高林 厚史
  • 新規手法による葉緑体タンパク質複合体の網羅的検出
    文部科学省:若手研究 (B)
    研究期間 : 2011年 -2013年 
    代表者 : 高林 厚史
  • バイオインフォマティクスの手法を用いたクロロフィル代謝経路の確立
    文部科学省:若手研究 (スタートアップ)→研究活動スタート支援
    研究期間 : 2009年 -2010年 
    代表者 : 高林 厚史

教育活動情報

主要な担当授業

  • 環境応答システム科学特論
    開講年度 : 2018年
    課程区分 : 修士課程
    開講学部 : 生命科学院
    キーワード : 光合成、クロロフィル代謝


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