Updated on 2024/03/29

写真a

 
KATO Akira
 
Organization
Academic Assembly Institute of Science and Technology CHIKYU SEIBUTSU KAGAKU KEIRETU Associate Professor
Graduate School of Science and Technology Life and Food Sciences Associate Professor
Faculty of Science Department of Science Associate Professor
Title
Associate Professor
External link

Degree

  • 博士(理学) ( 1995.3   総合研究大学院大学 )

Research Interests

  • Plant physiology

  • 植物生理学

Research Areas

  • Life Science / Plant molecular biology and physiology

Research History (researchmap)

  • - 新潟大学理学部 助教授

    2000

      More details

  • - Faculty of Science, Niigata University Associate professor

    2000

      More details

  • Niigata University

    1998 - 1999

      More details

  • Niigata University   Faculty of Science

    1998 - 1999

      More details

  • JSPS research associate

    1998

      More details

  • 未来開拓推進事業 リサーチアソシエイト

    1998

      More details

  • NIBB post doctoral fellow

    1995 - 1998

      More details

  • National Institute for Basic Biology

    1995 - 1998

      More details

▶ display all

Research History

  • Niigata University   Faculty of Science Department of Science   Associate Professor

    2017.4

  • Niigata University   Graduate School of Science and Technology Life and Food Sciences   Associate Professor

    2004.4

  • Niigata University   Graduate School of Science and Technology Life and Food Sciences   Associate Professor

    2004.4

  • Niigata University   Faculty of Science Department of Biology   Associate Professor

    2004.4 - 2017.3

  • Niigata University   Faculty of Science   Research Assistant

    1998.9 - 1999.12

Education

  • The Graduate University for Advanced Studies   Graduate School, Division of Life Science

    - 1995

      More details

  • The Graduate University for Advanced Studies   生命科学研究科   分子生物機構論

    - 1995

      More details

    Country: Japan

    researchmap

Professional Memberships

 

Papers

  • Functional Transformation of Plant Peroxisomes

    Makoto Hayashi, Kanako Toriyama, Maki Kondo, Akira Kato, Shoji Mano, Luigi De Bellis, Yasuko Hayashi-Ishimaru, Katsushi Yamaguchi, Hiroshi Hayashi, Mikio Nishimura

    Cell Biochemistry and Biophysics   32   295 - 304   2000

     More details

    Publishing type:Research paper (scientific journal)  

    Peroxisomes in higher plant cells are known to differentiate into at least three different classes, namely, glyoxysomes, leaf peroxisomes, and unspecialized peroxisomes, depending on the cell types. In germinating fatty seedlings, glyoxysomes that first appear in the etiolated cotyledonary cells are functionally transformed into leaf peroxisomes during greening. Subsequently, the organelles are transformed back into glyoxysomes during senescence of the cotyledons. Flexibility of function is a distinct feature of plant peroxisomes. This article briefly describes recent studies of the regulatory mechanisms involved in the changes of the function of plant peroxisomes.

    DOI: 10.1385/CBB:32:1-3:295

    Scopus

    PubMed

    researchmap

  • Microbody defective mutants of arabidopsis

    Mikio Nishimura, Makoto Hayashi, Kanako Toriyama, Akira Kato, Shoji Mano, Katsushi Yamaguchi, Maki Kondo, Hiroshi Hayashi

    Journal of Plant Research   111 ( 2 )   329 - 332   1998.6

     More details

    Publishing type:Research paper (scientific journal)  

    In germinating fatty seedlings, microbodies are differentiated to leaf peroxisomes from glyoxysomes during greening, and then transformed to glyoxysomes from leaf peroxisomes during senescence. These transformations of microbodies are regulated at various level, such as gene expression, splicing of the mRNA and degradation of microbody proteins. In order to clarify the regulatory mechanisms underlying these transformations of microbodies, we tried to obtain glyoxysome-deficient mutants of Arabidopsis. We screened 2,4-dichlorophenoxybutyric acid (2,4-DB) mutants of Arabidopsis which have defects in glyoxysomal fatty acid β-oxidation. Four mutants can be classified as carrying alleles at three independent loci, which we designated ped1, ped2, and ped3, respectively (where ped stands for peroxisome defective). The characteristics of these ped mutants are described.

    DOI: 10.1007/bf02512192

    Scopus

    researchmap

  • Functional transformation of microbodies in higher plant cells

    Mikio Nishimura, Makoto Hayashi, Akira Kato, Katsushi Yamaguchi, Shoji Mano

    Cell Structure and Function   21 ( 5 )   387 - 393   1996.10

     More details

    Publishing type:Research paper (international conference proceedings)  

    In germinating fatty seedlings, microbodies are functionally transformed to leaf peroxisomes from glyoxysomes during greening, and then converted to glyoxysomes from leaf peroxisomes during senescence. Immunocytochemical studies revealed that glyoxysomes can exchange directly into leaf peroxisomes during greening and leaf peroxisomes are once again directly converted to glyoxysomes during senescence. The reversible transformations of microbodies are regulated at various levels, such as gene expression, splicing of the mRNA and degradation of microbody proteins. The regulatory mechanisms underlying this organelle differentiation are described.

    DOI: 10.1247/csf.21.387

    Scopus

    PubMed

    researchmap

Books

  • 標識を付けたタンパク質の植物での発現,シロイヌナズナでの発現と細胞内局在性の解析

    秀潤社,植物細胞工学シリ-ズ「植物のタンパク質実験プロトコ-ル」  1998 

     More details

MISC

  • Chaperone and Protease Functions of LON Protease 2 Modulate the Peroxisomal Transition and Degradation with Autophagy

    Shino Goto-Yamada, Shoji Mano, Chihiro Nakamori, Maki Kondo, Ryuichi Yamawaki, Akira Kato, Mikio Nishimura

    Plant and Cell Physiology   55 ( 3 )   482 - 496   2014.3

     More details

    Publisher:Oxford University Press (OUP)  

    DOI: 10.1093/pcp/pcu017

    researchmap

  • The role of A rabidopsis thaliana NAR 1, a cytosolic iron–sulfur cluster assembly component, in gametophytic gene expression and oxidative stress responses in vegetative tissue

    Miyuki Nakamura, Diana Mihaela Buzas, Akira Kato, Masahiro Fujita, Nori Kurata, Tetsu Kinoshita

    New Phytologist   199 ( 4 )   925 - 935   2013.9

  • Functional analysis of a peroxisome-localized protease-like protein in Arabidopsis

    M Morohashi, T Hayakawa, S Mano, M Nishimura, A Kato

    PLANT AND CELL PHYSIOLOGY   46   S229 - S229   2005

     More details

    Language:English   Publishing type:Research paper, summary (international conference)   Publisher:OXFORD UNIV PRESS  

    Web of Science

    researchmap

  • Characterization of a peroxisome-localized small heat shock protein in Arabidopsis.

    K Kanou, S Mano, M Nishimura, A Kato

    PLANT AND CELL PHYSIOLOGY   45   S229 - S229   2004

     More details

    Language:English   Publishing type:Research paper, summary (international conference)   Publisher:OXFORD UNIV PRESS  

    Web of Science

    researchmap

  • Characterization of a small HSP like protein that has a peroxisomal targeting signal

    K Kanou, S Mano, M Nishimura, A Kato

    PLANT AND CELL PHYSIOLOGY   44   S106 - S106   2003

     More details

    Language:English   Publishing type:Research paper, summary (international conference)   Publisher:OXFORD UNIV PRESS  

    Web of Science

    researchmap

  • Molecular characterization of an Arabidopsis acyl-coenzyme A synthetase localized on glyoxysomal membranes

    H Hayashi, L De Bellis, Y Hayashi, K Nito, A Kato, M Hayashi, Hara-Nishimura, I, M Nishimura

    PLANT PHYSIOLOGY   130 ( 4 )   2019 - 2026   2002.12

     More details

    Language:English   Publisher:AMER SOC PLANT BIOLOGISTS  

    In higher plants, fat-storing seeds utilize storage lipids as a source of energy during germination. To enter the beta-oxidation pathway, fatty acids need to be activated to acyl-coenzyme As (CoAs) by the enzyme acyl-CoA synthetase (ACS; EC 6.2.1.3). Here, we report the characterization of an Arabidopsis cDNA clone encoding for a glyoxysomal acyl-CoA synthetase designated AtLACS6. The cDNA sequence is 2,106 bp long and it encodes a polypeptide of 701 amino acids with a calculated molecular mass of 76,617 D. Analysis of the amino-terminal sequence indicates that acyl-CoA synthetase is synthesized as a larger precursor containing a cleavable amino-terminal presequence so that the mature polypeptide size is 663 amino acids. The presequence shows high similarity to the typical PTS2 (peroxisomal targeting signal 2). The AtLACS6 also shows high amino acid identity to prokaryotic and eukaryotic fatty acyl-CoA synthetases. Immunocytochemical and cell fractionation analyses indicated that the AtLACS6 is localized on glyoxysomal membranes. AtLACS6 was overexpressed in insect cells and purified to near homogeneity. The purified enzyme is particularly active on long-chain fatty acids (C16:0). Results from immunoblot analysis revealed that the expression of both AtLACS6 and beta-oxidation enzymes coincide with fatty acid degradation. These data suggested that AtLACS6 might play a regulatory role both in fatty acid import into glyoxysomes by making a complex with other factors, e.g. PMP70, and in fatty acid beta-oxidation activating the fatty acids.

    DOI: 10.1104/pp.012955

    Web of Science

    researchmap

  • Molecular characterization of an Arabidopsis acyl-coenzyme A synthetase localized on glyoxysomal membranes

    H Hayashi, L De Bellis, Y Hayashi, K Nito, A Kato, M Hayashi, Hara-Nishimura, I, M Nishimura

    PLANT PHYSIOLOGY   130 ( 4 )   2019 - 2026   2002.12

     More details

    Language:English   Publisher:AMER SOC PLANT BIOLOGISTS  

    In higher plants, fat-storing seeds utilize storage lipids as a source of energy during germination. To enter the beta-oxidation pathway, fatty acids need to be activated to acyl-coenzyme As (CoAs) by the enzyme acyl-CoA synthetase (ACS; EC 6.2.1.3). Here, we report the characterization of an Arabidopsis cDNA clone encoding for a glyoxysomal acyl-CoA synthetase designated AtLACS6. The cDNA sequence is 2,106 bp long and it encodes a polypeptide of 701 amino acids with a calculated molecular mass of 76,617 D. Analysis of the amino-terminal sequence indicates that acyl-CoA synthetase is synthesized as a larger precursor containing a cleavable amino-terminal presequence so that the mature polypeptide size is 663 amino acids. The presequence shows high similarity to the typical PTS2 (peroxisomal targeting signal 2). The AtLACS6 also shows high amino acid identity to prokaryotic and eukaryotic fatty acyl-CoA synthetases. Immunocytochemical and cell fractionation analyses indicated that the AtLACS6 is localized on glyoxysomal membranes. AtLACS6 was overexpressed in insect cells and purified to near homogeneity. The purified enzyme is particularly active on long-chain fatty acids (C16:0). Results from immunoblot analysis revealed that the expression of both AtLACS6 and beta-oxidation enzymes coincide with fatty acid degradation. These data suggested that AtLACS6 might play a regulatory role both in fatty acid import into glyoxysomes by making a complex with other factors, e.g. PMP70, and in fatty acid beta-oxidation activating the fatty acids.

    DOI: 10.1104/pp.012955

    Web of Science

    researchmap

  • Distribution and characterization of peroxisomes in arabidopsis by visualization with GFP: Dynamic morphology and actin-dependent movement

    S Mano, C Nakamori, M Hayashi, A Kato, M Kondo, M Nishimura

    PLANT AND CELL PHYSIOLOGY   43 ( 3 )   331 - 341   2002.3

     More details

    Language:English   Publisher:OXFORD UNIV PRESS  

    Peroxisomes were visualized in living cells of various tissues in transgenic Arabidopsis by green fluorescent protein (GFP) through the addition of the peroxisomal targeting signal 1 (PTS1) or PTS2. The observation using confocal laser scanning microscopy revealed that the GFP fluorescence signals were detected as spherical spots in all cells of two kinds of transgenic plants. Immunoelectron microscopic analysis using antibodies against the peroxisomal marker protein, catalase, showed the presence of GFP in peroxisomes, confirming that GFP was correctly transported into peroxisomes by PTS1 or PTS2 pathways. It has been also revealed that peroxisomes are motile organelles whose movement might be caused by cytoplasmic flow. The movement of peroxisomes was more prominent in root cells than that in leaves, and divided into two categories: a relatively slow, random, vibrational movement and a rapid movement. Treatment with anti-actin and anti-tubulin drugs revealed that actin filaments involve in the rapid movement of peroxisomes. Moreover, abnormal large peroxisomes are present as clusters at the onset of germination, and these clusters disappear in a few days. Interestingly, tubular peroxisomes were also observed in the hypocotyl. These findings indicate that the shape, size, number and movement of peroxisomes in living cells are dynamic and changeable rather than uniform.

    DOI: 10.1093/pcp/pcf037

    Web of Science

    researchmap

  • Distribution and characterization of peroxisomes in arabidopsis by visualization with GFP: Dynamic morphology and actin-dependent movement

    S Mano, C Nakamori, M Hayashi, A Kato, M Kondo, M Nishimura

    PLANT AND CELL PHYSIOLOGY   43 ( 3 )   331 - 341   2002.3

     More details

    Language:English   Publisher:OXFORD UNIV PRESS  

    Peroxisomes were visualized in living cells of various tissues in transgenic Arabidopsis by green fluorescent protein (GFP) through the addition of the peroxisomal targeting signal 1 (PTS1) or PTS2. The observation using confocal laser scanning microscopy revealed that the GFP fluorescence signals were detected as spherical spots in all cells of two kinds of transgenic plants. Immunoelectron microscopic analysis using antibodies against the peroxisomal marker protein, catalase, showed the presence of GFP in peroxisomes, confirming that GFP was correctly transported into peroxisomes by PTS1 or PTS2 pathways. It has been also revealed that peroxisomes are motile organelles whose movement might be caused by cytoplasmic flow. The movement of peroxisomes was more prominent in root cells than that in leaves, and divided into two categories: a relatively slow, random, vibrational movement and a rapid movement. Treatment with anti-actin and anti-tubulin drugs revealed that actin filaments involve in the rapid movement of peroxisomes. Moreover, abnormal large peroxisomes are present as clusters at the onset of germination, and these clusters disappear in a few days. Interestingly, tubular peroxisomes were also observed in the hypocotyl. These findings indicate that the shape, size, number and movement of peroxisomes in living cells are dynamic and changeable rather than uniform.

    DOI: 10.1093/pcp/pcf037

    Web of Science

    researchmap

  • Molecular analysis of a protease that is localized in peroxisomes

    F Meguro, S Mano, M Nishimura, A Kato

    PLANT AND CELL PHYSIOLOGY   43   S35 - S35   2002

     More details

    Language:English   Publishing type:Research paper, summary (international conference)   Publisher:OXFORD UNIV PRESS  

    Web of Science

    researchmap

  • Transport of Peroxisomal Proteins Synthesized as Large Precursors in Plants

    Akira Kato, Makoto Hayashi, Maki Kondo, Mikio Nishimura

    Cell Biochemistry and Biophysics   32 ( 269-275 )   269 - 275   2000

     More details

    Language:English   Publisher:Humana Press  

    Plant peroxisomes contain at least four proteins, namely, citrate synthase, malate dehydrogenase, long-chain acyl-CoA oxidase, and 3-ketoacyl-CoA thiolase, which are synthesized as large precursors with an N-terminal cleavable presequence. Each presequence has a conserved domain (R[I/L/Q]-X5-HL) that is homologous to peroxisomal targeting signal 2 from mammals and yeasts. In addition, a cysteine residue is found at the C-terminal ends of the presequences, whose function has not yet been described. The authors analyzed the function of the presequences and the conserved amino acids using transgenic Arabidopsis plants, which accumulate β-glucuronidase carrying the presequence of the peroxisomal proteins from plants. Immunological and immunocytochemical studies on the transgenic plants showed that a conserved sequence in the extrapeptides is essential for targeting to peroxisomes, and a cysteine residue at the cleavage site is involved in the processing of the presequence. These results suggest that the presequences of the peroxisomal proteins function as targeting signals, and are necessary for the recognition of the processing.

    DOI: 10.1385/CBB:32:1-3:269

    Scopus

    PubMed

    researchmap

  • Transport of peroxisomal profeins that synthesized as largy precursors in plauts.

    Cell Biochemistry and Biophysics   32 ( 269-275 )   2000

     More details

  • Oligomeric proteins containing N-terminal targeting signals are imported into peroxisomes in transgenic Arabidopsis

    A Kato, M Hayashi, M Nishimura

    PLANT AND CELL PHYSIOLOGY   40 ( 6 )   586 - 591   1999.6

     More details

    Language:English   Publisher:JAPANESE SOC PLANT PHYSIOLOGISTS  

    Employing transgenic Arabidopsis plants, we analyzed the mechanism for the translocation of peroxisomal proteins from the cytosol into the matrix that is mediated by the N-terminal targeting signal. A hybrid Arabidopsis variety was generated which accumulates two kinds of originally bacterial proteins, P-glucuronidase (GUS) and a GUS chimeric protein designated as CS-Delta C42-GUS, that carries the N-terminal targeting signal for glyoxysomal citrate synthase. Because the CS-Delta C42-GUS is targeted to peroxisomes but had never been observed to be processed to produce the mature protein, it can be distinguished from the GUS protein by its molecular size. Cell fractionation analyses showed that the native GUS protein, although lacking the targeting signal, was co-localized with the CS-Delta C42-GUS protein in the peroxisomes of the hybrid plant. It is suggested that the native GUS protein forms oligomeric structures with the peroxisome-targeted chimeric proteins and can therefore be transported into peroxisomes. Sucrose density gradient centrifugation revealed that the native GUS and the chimeric GUS indeed are present both as a dimer and a tetramer in the Arabidopsis hybrid variety.

    Web of Science

    researchmap

  • Molecular characterization of a glyoxysomal long chain acyl-CoA oxidase that is synthesized as a precursor of higher molecular mass in pumpkin

    H Hayashi, L De Bellis, K Yamaguchi, A Kato, M Hayashi, M Nishimura

    JOURNAL OF BIOLOGICAL CHEMISTRY   273 ( 14 )   8301 - 8307   1998.4

     More details

    Language:English   Publisher:AMER SOC BIOCHEMISTRY MOLECULAR BIOLOGY INC  

    A cDNA clone for pumpkin acyl-CoA oxidase (EC 1.3.3.6; ACOX) was isolated from a lambda gt11 cDNA library constructed from poly(A)(+) RNA extracted from etiolated cotyledons, The inserted cDNA clone contains 2313 nucleotides and encodes a polypeptide of 690 amino acids, Analysis of the amino-terminal sequence of the protein indicates that the pumpkin acyl-CoA oxidase protein is synthesized as a larger precursor containing a cleavable amino-terminal presequence of 45 amino acids. This presequence shows high similarity to the typical peroxisomal targeting signal (PTS2). Western blot analysis following cell fractionation in a sucrose gradient revealed that ACOX is localized in glyoxysomes, A partial purification of ACOX from etiolated pumpkin cotyledons indicated that the ACOX cDNA codes for a long chain acyl-CoA oxidase, The amount of ACOX increased and reached to the maximum activity by day 5 of germination but decreased about 4-fold on the following days during the subsequent microbody transition from glyoxysomes to leaf peroxisomes. By contrast, the amount of mRNA was already high at day 1 of germination, increased by about 30% at day 3, and faded com pletely by day 7. These data indicated that the expression pattern of ACOX was very similar to that of the glyoxysomal enzyme 3-ketoacyl-CoA thiolase, another marker enzyme of the beta-oxidation spiral, during germination and suggested that the expression of each enzyme of P-oxidation is coordinately regulated.

    DOI: 10.1074/jbc.273.14.8301

    Web of Science

    PubMed

    CiNii Article

    researchmap

  • Glyoxysomal malate dehydrogenase in pumpkin: Cloning of a cDNA and functional analysis of its presequence

    A Kato, Y Takeda-Yoshikawa, M Hayashi, M Kondo, Hara-Nishimura, I, M Nishimura

    PLANT AND CELL PHYSIOLOGY   39 ( 2 )   186 - 195   1998.2

     More details

    Language:English   Publisher:JAPANESE SOC PLANT PHYSIOLOGISTS  

    Glyoxysomal malate dehydrogenase (gMDH) is an enzyme of the glyoxylate cycle that participates in degradation of storage oil, We have cloned a cDNA for gMDH from etiolated pumpkin cotyledons that encodes a polypeptide consisting of 356 amino acid residues, The nucleotide and N-terminal amino acid sequences revealed that gMDH is synthesized as a precursor with an N-terminal extrapeptide, The N-terminal presequence of 36 amino acid residues contains two regions homologous to those of other microbody proteins, which are also synthesized as large precursors, To investigate the functions of the N-terminal presequence of gMDH, we generated transgenic Arabidopsis that expressed a chimeric protein consisting of beta-glucuronidase and the N-terminal region of gMDH. Immunological and immunocytochemical studies revealed that the chimeric protein was imported into microbodies such as glyoxysomes and leaf peroxisomes and was then subsequently processed, Site-directed mutagenesis studies showed that the conserved amino acids in the N-terminal presequence, Arg-10 and His-17, function as recognition sites for the targeting to plant microbodies, and Cys-36 in the presequence is responsible for its processing. These results correspond to those from the analyses of glyoxysomal citrate synthase (gCS), which was also synthesized as a large precursor, suggesting that common mechanisms that can recognize the targeting or the processing of gMDH and gCS function in higher plant cells.

    Web of Science

    researchmap

  • Protein import to microbodies in transgenic Arabidopsis.

    A Kato, M Hayashi, M Kondo, M Nishimura

    PLANT PHYSIOLOGY   114 ( 3 )   1197 - 1197   1997.7

     More details

    Language:English   Publishing type:Research paper, summary (international conference)   Publisher:AMER SOC PLANT PHYSIOLOGISTS  

    Web of Science

    researchmap

  • RECOGNITION OF THE N-TERMINAL TARGETING SIGNAL OF GLYOXYSOMAL CITRATE SYNTHASE PRECURSOR IN TRANSGENIC ARABIDOPSIS

    KATO Akira, HAYASHI Makoto, NISHIMURA Mikio

    38   s123   1997.3

     More details

  • cDNA CLONING AND EXPRESSION OF A GENE FOR THE ENZYMES OF β-OXIDATION CYCLE IN PUMPKIN COTYLEDONS

    HAYASHI Hiroshi, KATO Akira, HAYASHI Makoto, NISHIMURA Mikio

    38   s50   1997.3

     More details

  • Targeting and processing of a chimeric protein with the N-terminal presequence of the precursor to glyoxysomal citrate synthase

    A Kato, M Hayashi, M Kondo, M Nishimura

    PLANT CELL   8 ( 9 )   1601 - 1611   1996.9

     More details

    Language:English   Publisher:AMER SOC PLANT PHYSIOLOGISTS  

    Glyoxysomal citrate synthase in pumpkin is synthesized as a precursor that has a cleavable presequence at its N-terminal end. To investigate the role of the presequence in the transport of the protein to the microbodies, we generated transgenic Arabidopsis plants that expressed beta-glucuronidase with the N-terminal presequence of the precursor to the glyoxysomal citrate synthase of pumpkin. Immunogold labeling and cell fractionation studies showed that the chimeric protein was transported into microbodies and subsequently was processed. The chimeric protein was transported to functionally different microbodies, such as glyoxysomes, leaf peroxisomes, and unspecialized microbodies. These observations indicated that the transport of glyoxysomal citrate synthase is mediated by its N-terminal presequence and that the transport system is functional in all plant microbodies. Site-directed mutagenesis of the conserved amino acids in the presequence caused abnormal targeting and inhibition of processing of the chimeric protein, suggesting that the conserved amino acids in the presequence are required for recognition of the target or processing.

    DOI: 10.1105/tpc.8.9.1601

    Web of Science

    PubMed

    CiNii Article

    researchmap

  • Transport of chimeric proteins that contain a carboxy-terminal targeting signal into plant microbodies

    M Hayashi, M Aoki, A Kato, M Kondo, M Nishimura

    PLANT JOURNAL   10 ( 2 )   225 - 234   1996.8

     More details

    Language:English   Publisher:BLACKWELL SCIENCE LTD  

    Malate synthase is a glyoxysome-specific enzyme. The carboxy-terminal tripeptide of the enzyme is Ser-Arg-Leu (SRL), which is known to function as a peroxisomal targeting signal in mammalian cells. To analyze the function of the carboxy-terminal amino acids of pumpkin malate synthase in plant cells, a chimeric gene was con structed that encoded a fusion protein which consisted of beta-glucuronidase and the carboxyl terminus of the enzyme. The fusion protein was expressed and accumulated in transgenic Arabidopsis that had been transformed with the chimeric gene. Immunocytochemical analysis of the transgenic plants revealed that the carboxy-terminal five amino acids of pumpkin malate synthase were sufficient for transport of the fusion protein into glyoxysomes in etiolated cotyledons, into leaf peroxisomes in green cotyledons and in mature leaves, and into unspecialized microbodies in roots, although the fusion protein was no longer transported into microbodies when SRL at the carboxyl terminus was deleted. Transport of proteins into glyoxysomes and leaf peroxisomes was also observed when the carboxy-terminal amino acids of the fusion protein were changed from SRL to SKL, SRM, ARL or PRL. The results suggest that tripeptides with S, A or P at the -3 position, K or R at the -2 position, and L or M at the carboxyl terminal position can function as a targeting signal for three kinds of plant microbody.

    DOI: 10.1046/j.1365-313X.1996.10020225.x

    Web of Science

    researchmap

  • cDNA cloning and expression of a gene for 3-ketoacyl-CoA thiolase in pumpkin cotyledons

    A Kato, M Hayashi, Y Takeuchi, M Nishimura

    PLANT MOLECULAR BIOLOGY   31 ( 4 )   843 - 852   1996.7

     More details

    Language:English   Publisher:KLUWER ACADEMIC PUBL  

    A cDNA clone for 3-ketoacyl-CoA thiolase (EC 2.3.1.16) was isolated from a lambda gt11 cDNA library constructed from the poly(A)(+) RNA of etiolated pumpkin cotyledons. The cDNA insert contained 1682 nucleotides and encoded 461 amino acid residues. A study of the expression in vitro of the cDNA and analysis of the amino-terminal sequence of the protein indicated that pumpkin thiolase is synthesized as a precursor which has a cleavable amino-terminal presequence of 33 amino acids. The amino-terminal presequence was highly homologous to typical amino-terminal et proteins to microbodies. Immunoblot analysis showed that the amount of thiolase increased markedly during germination but decreased dramatically during the light-inducible transition of microbodies from glyoxysomes to leaf peroxisomes. By contrast, the amount of mRNA increased temporarily during the early stage of germination. In senescing cotyledons, the levels of the thiolase mRNA and protein increased again with the reverse transition of microbodies from leaf peroxisomes to glyoxysomes, but the pattern of accumulation of the protein was slightly different from that of malate synthase. These results indicate that expression of the thiolase is regulated in a similar manner to that of other glyoxysomal enzymes, such as malate synthase and citrate synthase, during seed germination and post-germination growth. By contrast, during senescence, expression of the thiolase is regulated in a different manner from that of other glyoxysomal enzymes.

    DOI: 10.1007/BF00019471

    Web of Science

    PubMed

    CiNii Article

    researchmap

  • THE N-TERMINAL REGION OF GLYOXYSOMAL MALATE DEHYDROGENASE FROM PUMPKIN FUNCTION IN TARGETING TO MICROBODIES

    KATO Akira, HAYASHI Makoto, KONDO Maki, NISHIMURA Mikio

    37   101 - 101   1996.3

     More details

  • MOLECULAR CHARACTERIZATION OF A GLYOXYSOMAL CITRATE SYNTHASE THAT IS SYNTHESIZED AS A PRECURSOR OF HIGHER MOLECULAR-MASS IN PUMPKIN

    A KATO, M HAYASHI, H MORI, M NISHIMURA

    PLANT MOLECULAR BIOLOGY   27 ( 2 )   377 - 390   1995.1

     More details

    Language:English   Publisher:SPRINGER  

    A cDNA clone for glyoxysomal citrate synthase (gCS) was isolated from a lambda gt11 cDNA library prepared from etiolated pumpkin cotyledons. The cDNA of 1989 bp consisted of a 1548 bp open reading frame that encoded 516 amino acid residues. The deduced amino acid sequence of gCS did not have a typical peroxisomal targeting signal at its carboxyl terminal. A study of expression in vitro of the cDNA and an analysis of the amino-terminal sequence of the citrate synthase indicated that gCS is synthesized as a larger precursor that has a cleavable amino-terminal presequence of 43 amino acids. The predicted amino-terminal sequence of pumpkin gCS was highly homologous to those of other microbody enzymes, such as 3-ketoacyl-CoA thiolase of rat and malate dehydrogenase of watermelon that are also synthesized as precursors of higher molecular mass.
    Immunoblot analysis showed that the level of gCS protein increased markedly during germination and decreased rapidly during the light-induced transition of microbodies from glyoxysomes to leaf peroxisomes. By contrast, the level of mRNA for gCS reached a maximum earlier than that of the protein and declined even in darkness. The level of the mRNA was low during the microbody transition. These results indicate that the accumulation of the gCS protein does not correspond to that of the mRNA and that degradation of gCS is induced during the microbody transition, suggesting that post-transcriptional regulation plays an important role in the microbody transition.

    Web of Science

    researchmap

  • CHARACTERIZATION OF INTRAVACUOLAR PIGMENTED STRUCTURES IN ANTHOCYANIN-CONTAINING CELLS OF SWEET-POTATO SUSPENSION-CULTURES

    M NOZUE, H KUBO, M NISHIMURA, A KATO, C HATTORI, N USUDA, T NAGATA, H YASUDA

    PLANT AND CELL PHYSIOLOGY   34 ( 6 )   803 - 808   1993.9

     More details

    Language:English   Publisher:JAPANESE SOC PLANT PHYSIOLOGISTS  

    Intravacuolar pigmented structures occurred in anthocyanin-producing cultured cells of sweet potato (Ipomoea batatas) were characterized. Formation of the pigmented structures in sweet potato cells was induced by transfer of callus cultured in 2,4-D containing agar medium into 2,4-D free liquid medium under continuous illumination. These structures were found in the vacuoles. The pigmented structures were isolated from the protoplasts by precipitation in 60% (w/w) sucrose after centrifugation. Electron microscopic observations of the anthocyanin-containing cultured cells showed these structures had neither membrane boundary nor internal structures, and were found as strongly osmiophilic globules in vacuoles. Numerous small osmiophilic globules were observed in central vacuoles at the early stage of anthocyanin accumulation, but not found in cytoplasm. Similar pigmented structures in vacuoles were also formed by treatment with neutral red. These observations indicate that these pigmented structure is the high density and insoluble globules highly concentrated with anthocyanin, which was synthesized in cytoplasm and transported to the central vacuoles.

    Web of Science

    researchmap

▶ display all

Works

  • 植物にみる環境刺激の受容から応答に至る過程の研究

    2002

     More details

  • Joint Research of sensing and response of plants for environmental stimuli

    2002

     More details

  • 植物ペルオキシソームに局在するsmall HSP様タンパク質の機能.

    2000

     More details

  • シロイヌナズナによる植物B酸化系の形態形成およびストレス応答への関与

    2000

     More details

  • Joint Research of Functions of small HSP-like proteins localized in plant peroxisomes.

    2000

     More details

  • Joint research on participation of B-oxidation pathway on morphogenesis and stress-response of plants.

    2000

     More details

  • ペルオキシソーム局在型低分子量熱ショックタンパク質ホモログの構造と機能

    1998
    -
    2001

     More details

  • Joint research on structure and functions of peroxisome-localized small HSP-like proteins.

    1998
    -
    2001

     More details

▶ display all

Research Projects

  • ペルオキシソームにおける分子シャペロンの機能

    Grant number:19039011

    2007 - 2008

    System name:科学研究費助成事業

    Research category:特定領域研究

    Awarding organization:日本学術振興会

    加藤 朗

      More details

    Grant amount:\4600000 ( Direct Cost: \4600000 )

    高等植物のペルオキシソームには複数の分子シャペロン様タンパク質が局在するが, それらの生理的機能は不明である。申請者は材料としてシロイヌナズナとその突然変異体を用い, 分子生物学, 生化学, 分子遺伝学の手法によって3つのシャペロン様ペルオキシソームタンパク質(HSP15.7, Lon2, Deg15)の生理機能解析を行った。その結果, 次のような結果が得られた。(1) 低分子量熱ショックタンパック質ホモログであるHSP15.7を欠損した変異シロイヌナズナでは, 42℃の高温ストレスで光合成活性が低下したことから, HSP15.7は高温下における光呼吸活性の維持に関与ずると推定された。(2) 分子シャペロン活性を持つ多機能プロテアーゼであるLon2を欠損した変異シロイヌナズナでは, 種子発芽時に2, 4-DB耐性を示すことがわかった。これはLon2が発芽時におけるβ酸化系の機能発現に関与することを意味する。(3) 大腸菌DegプロテアーゼのホモログであるDeg15を欠損したシロイヌナズナでは, ペルオキシソームタンパク質の高分子量前駆体が蓄積した。また発芽初期において, 脂肪代謝系酵素群の発現が転写ならびに翻訳レベルで変動し, さらにβ酸化系活性が低下した。これら, Deg15がペルオキシソームタンパク質の成熟化を担うプロセシングプロテアーゼであること, 発芽初期における脂肪酸代謝系酵素群の遺伝子発現制御に関与することを示している。以上の結果から, 高等植物におけるペルオキシソームの機能発現, 機能維持には複数のシャペロン様タンパク質が関与することが明らかとなった。

    researchmap

  • ペルオキシソームにおける環境ストレス応答および耐性の分子機構

    Grant number:17051010

    2005 - 2006

    System name:科学研究費助成事業

    Research category:特定領域研究

    Awarding organization:日本学術振興会

    加藤 朗

      More details

    Grant amount:\6000000 ( Direct Cost: \6000000 )

    1.シロイヌナズナ変異体によるAtHSP15.7の機能解析
    ペルオキシソームに局在する分子シャペロンAtHSP15.7の機能を明らかにするためにT-DNA挿入変異株hsp15.7の解析を行った。その結果,(1)hsp-15.7では光呼吸系酵素であるHPR活性が低い一方,GGT活性が高い傾向が認められた。しかし(2)高温ストレス下における脂肪代謝活性やカタラーゼ並びに光呼吸関連酵素の活性変動パターンに,野生型との大きな差異は認められなかった。また(3)カタラーゼ阻害剤である3-アミノトリアゾール(3-AT)で緑葉を処理して弱光下培養すると,hsp-15.7のみにおいて緑葉の枯死が誘導された。これは,AtHSP15.7がペルオキシソームにおける酸化ストレスの低減に寄与する可能性を示唆する。
    2.シロイヌナズナ変異体によるAtLon1lの機能解析
    RNA干渉によってAtLon1の発現抑制株lonliを作製した。発芽過程における塩ストレス感受性を野生型と比較したが,発芽のわずかな遅延以外に外観および生育過程における大きな差異は認められなかった。現在,lonliにおけるペルオキシソームタンパク質の網羅的解析を試みている。
    3.Lon1の普遍性
    AtLon1と高い相同性を示し,塩ストレスに応答するイネプロテアーゼホモログ遺伝子OsLon1を同定した。組換えタンパク質によるin virto解析の結果,OsLon1のLonドメインは,AtLon1のLonドメイン同様,シャペロン活性を有することが明らかになった。またOsLon1のLonドメインを強制発現させた組換えイネ培養細胞では,塩ストレスによる細胞死が抑制される傾向が観察された。
    4.新奇シャペロン様ペルオキシソームタンパク質の同定
    分子シャペロンドメインを持つ新奇ペルオキシソームタンパク質としてAtDegQを同定した。

    researchmap

  • Flowering induced by DNA demethylation in Perilla frutescens var. crispa

    Grant number:15570032

    2003 - 2004

    System name:Grants-in-Aid for Scientific Research

    Research category:Grant-in-Aid for Scientific Research (C)

    Awarding organization:Japan Society for the Promotion of Science

    WADA Kiyotoshi, IWASAKI Toshisuke, KATO Akira

      More details

    Grant amount:\3600000 ( Direct Cost: \3600000 )

    The long-lasting effect of low temperature in vernalization is regulated by DNA demethylation. In photoperiodic flowering, the effect of photoperiodic cycle disappears if the plants were moved to unsuitable photoperiodic condition. However, a short-day plant, Perilla frutescens var. crispa is an exception. Once-induced flowering state of lasts long in this species. This suggests a possible involvement of DNA demethylation in the flowering of P.frutescens. Accordingly, the possible involvement of DNA demethylation in the regulation of flowering in P.frutescens was studied.
    1.A DNA demethylating reagent, 5-azacytidine was treated to a short-day plant, Perilla frutescens var. crispa under a long-day condition. The treatment caused flowering, suggesting that DNA demethylation induced flowering.
    2.The genomic DNAs were isolated from the leaves, digested with a methylation-sensitive restriction enzyme, Hpa II, and then Southern hybridization analysis was performed with a probe, 25S-18S rDNA intergenic spacer region cloned from P.frutescens. Lower molecular fragments were detected only in the 5-azacytidine-treated plants. The result indicates that the 5-azacytidine treatment caused demethylation of the genomic DNA.
    3.The treatment of shoot apical meristem was performed when the leaves at the fourth node were unfolded. Floral inflorescences were induced even at the lower leaf axils. Such an indirect may be caused by flowering stimulus transported from the leaves at the higher nodes which were affected by 5-azacytidine.
    4.The treatment with 5-azacytidine induced flowering also in Silene armeria. of which flowering state lasted Ions The induction of flowering may be correlated with the long-lasting memory of photoperiodic effect.

    researchmap

  • 高等植物におけるペルオキシソーム及びペルオキシソームタンパク質の新奇な機能の探索

    Grant number:13740454

    2001 - 2002

    System name:科学研究費助成事業

    Research category:若手研究(B)

    Awarding organization:日本学術振興会

    加藤 朗

      More details

    Grant amount:\2100000 ( Direct Cost: \2100000 )

    1.ペルオキシソームに局在するプロテアーゼAtLon1の解析
    ペルオキシソームに局在する新奇なプロテアーゼとして同定したAtLon1タンパク質の変動および組織特異的蓄積をシロイヌナズナを用いて解析した。その結果AtLon1タンパク質は、発芽初期の芽生えで多く蓄積すること、光照射によって蓄積量が減少すること。ロゼット葉、茎生葉、茎では蓄積が見られないが、根、つぼみ、花ではわずかに蓄積することが確認された。現在、mRNAレベルでの発現解析を行っている。
    2.シロイヌナズナの新奇低分子量熱ショックタンパク質HSP15.7の同定
    ペルオキシソーム輸送シグナルPTS1を有する新奇な低分子量熱ショックタンパク質HSP15.7をシロイヌナズナにおいて同定した。HSP15.7の遺伝子発現は、熱ストレスによって誘導され、また花やつぼみで顕著であるという特徴がみられた。さらに組換えタンパク質を用いた実験から、HSP15.7は分子シャペロンとして機能する新奇なペルオキシソームタンパク質である可能性が強く示唆された。現在特異抗体を調製中であり、今後はHSP15.7の細胞内局在性、タンパク質レベルでの発現解析を行う予定である。
    3.シロイヌナズナトランスポゾン挿入変異体を用いたペルオキシソーム酵素遺伝子の機能解析
    理化学研究所から供与を受けたシロイヌナズナトランスポゾン挿入変異体を用いて、ペルオキシソーム酵素の遺伝子であるMFP2(多機能酵素;β酸化系酵素)、およびMDH5(リンゴ酸脱水素酵素;グリオキシル酸回路酵素)の機能解析を行った。両遺伝子の変異体であるmfp2およびmdh5の表現型解析、野生型におけるMFP2とMDH5の発現解析の結果、MFP2およびMDH5はともにシロイヌナズナの発芽初期における脂肪酸代謝に必須であることが示唆された。

    researchmap

  • マイクロボディに局在する低分子量熱ショック蛋白質ホモログHSP27の構造と機能

    Grant number:11740439

    1999 - 2000

    System name:科学研究費助成事業

    Research category:奨励研究(A)

    Awarding organization:日本学術振興会

    加藤 朗

      More details

    Grant amount:\2300000 ( Direct Cost: \2300000 )

    トマトホモログ遺伝子の単離と発現解析
    今年度はシロイヌナズナHSP27の相同クローンとして,トマト緑葉からLeMP26cDNAを単離し,その一次構造を明らかにし,さらに遺伝子発現パターンの解析を行った。単離したLeMP26cDNAから予想されるポリペプチドは238アミノ酸からなり,C末端側には低分子量熱ショックタンパク質に共通する保存配列(α-クリスタリンドメイン)が,N末端にはペルオキシソームへの輸送シグナルであるPTS2配列との相同配列が見い出された。また,25℃で育成したトマト緑葉に38℃の熱ショックを与え,熱ショック後のLeMP26mRNAの変動をRT-PCRで検討した。その結果,緑葉では既にmRNAの蓄積が観察されること,熱ショックによって急激にmRNA量が減少し,その後,時間の経過とともに発現量が回復することが明らかになった。これは,LeMP26が熱ショック応答遺伝子ではないことを示している。
    組換えHSP27タンパク質を用いた機能解析
    シロイヌナズナHSP27の組換えタンパク質を大腸菌で合成,精製し,in vitroにおける機能と分子構造の解析を試みた。しかし,成熟型HSP27タンパク質を大腸菌で合成することはできなかった。そこで,他の低分子量熱ショックタンパク質とのホモロジーが低いN末端側102アミノ酸を切除し,α-クリスタリンドメインを含むC末端側147アミノ酸のみからなる組換えタンパク質(ΔN-HSP27)を合成した。精製したΔN-HSP27の分子量をショ糖密度勾配遠心法によって推定したところ,約21kDであった。従ってΔN-HSP27は,溶液中ではモノマーとして存在すると考えられる。また精製標品のシャペロン活性を測定したが,タンパク質の熱変成を阻害する活性,変成タンパク質のrefoldingを促進する活性,いずれも検出されなかった。切除した領域の機能は全く不明であるので,今後は真核細胞発現系を使用して全長HSP27の合成を行い,改めて活性測定を行う予定である。

    researchmap

  • 植物におけるペルオキシソームの形成と機能発現

    1999

      More details

    Grant type:Competitive

    researchmap

  • Biogenesis and functions of peroxisomes in plants

    1999

      More details

    Grant type:Competitive

    researchmap

▶ display all

 

Teaching Experience

  • 生物学実習I

    2022
    Institution name:新潟大学

  • 生物学実習II

    2022
    Institution name:新潟大学

  • 生物学実験

    2022
    Institution name:新潟大学

  • 基礎生物科学実習I

    2022
    Institution name:新潟大学

  • 植物機能制御論II

    2022
    Institution name:新潟大学

  • 理学スタディ・スキルズ

    2022
    Institution name:新潟大学

  • 基礎生物科学実習II

    2022
    Institution name:新潟大学

  • 生物学基礎演習

    2022
    Institution name:新潟大学

  • 生命科学への招待(生物学学習法)

    2022
    Institution name:新潟大学

  • 植物生理学実習

    2021
    Institution name:新潟大学

  • 生物学基礎A

    2021
    Institution name:新潟大学

  • 自然科学基礎実験

    2021
    Institution name:新潟大学

  • 生物学基礎実習a

    2021
    Institution name:新潟大学

  • 生物学特論 III

    2021
    Institution name:新潟大学

  • 植物生理学特論II

    2021
    Institution name:新潟大学

  • 日本事情自然系A

    2021
    Institution name:新潟大学

  • 専門力アクティブ・ラーニング

    2019
    -
    2020
    Institution name:新潟大学

  • 課題研究II(生物学)

    2018
    Institution name:新潟大学

  • 遺伝学

    2018
    Institution name:新潟大学

  • 生物学基礎実習b

    2017
    Institution name:新潟大学

  • 生物学総合演習

    2017
    Institution name:新潟大学

  • 課題研究I(生物学)

    2017
    Institution name:新潟大学

  • 理学スタディ・スキルズ

    2017
    -
    2022
    Institution name:新潟大学

  • 自然科学基礎実験

    2017
    Institution name:新潟大学

  • 基礎植物学

    2017
    Institution name:新潟大学

  • 自然科学総論Ⅳ

    2017
    Institution name:新潟大学

  • 生物学特論II

    2017
    Institution name:新潟大学

  • 植物生理学実習

    2016
    -
    2019
    Institution name:新潟大学

  • 植物生理学特論Ⅱ

    2015
    -
    2017
    Institution name:新潟大学

  • 生物学-植物A-

    2015
    Institution name:新潟大学

  • 生物学-植物A-

    2014
    Institution name:新潟大学

  • 生物学特論Ⅴ

    2014
    Institution name:新潟大学

  • 植物機能制御論Ⅱ

    2014
    Institution name:新潟大学

  • 細胞生物学II

    2012
    Institution name:新潟大学

  • 課題研究I(生物学科)

    2012
    -
    2016
    Institution name:新潟大学

  • 生命・食料科学特定研究BⅠ

    2012
    -
    2015
    Institution name:新潟大学

  • 研究発表演習(中間発表)

    2012
    -
    2015
    Institution name:新潟大学

  • 文献詳読Ⅰ

    2012
    -
    2015
    Institution name:新潟大学

  • 生命・食料科学セミナーBⅠ

    2012
    -
    2015
    Institution name:新潟大学

  • 生命・食料科学セミナーBⅡ

    2012
    -
    2014
    Institution name:新潟大学

  • 文献詳読Ⅱ

    2012
    -
    2014
    Institution name:新潟大学

  • 生命・食料科学特定研究BⅡ

    2012
    -
    2014
    Institution name:新潟大学

  • 研究発表

    2012
    Institution name:新潟大学

  • 生物学特論V B

    2011
    Institution name:新潟大学

  • 生物学基礎演習

    2010
    -
    2022
    Institution name:新潟大学

  • 科学・技術と社会

    2010
    -
    2012
    Institution name:新潟大学

  • 日本事情自然系A

    2009
    -
    2021
    Institution name:新潟大学

  • 基礎細胞生物学

    2008
    Institution name:新潟大学

  • 植物生理学演習

    2007
    Institution name:新潟大学

  • 生命科学への招待(生物学学習法)

    2007
    -
    2022
    Institution name:新潟大学

  • 基礎生物科学実習I

    2007
    -
    2022
    Institution name:新潟大学

  • 生物学基礎A

    2007
    -
    2019
    Institution name:新潟大学

  • 生物学実験 I

    2007
    -
    2019
    Institution name:新潟大学

  • 課題研究II

    2007
    -
    2016
    Institution name:新潟大学

  • 生物化学実習

    2007
    -
    2015
    Institution name:新潟大学

  • 植物形態発生学実習

    2007
    -
    2015
    Institution name:新潟大学

  • オルガネラ分子生物学特論

    2007
    -
    2013
    Institution name:新潟大学

  • 植物細胞生物学特論

    2007
    -
    2013
    Institution name:新潟大学

  • 課題研究I

    2007
    -
    2011
    Institution name:新潟大学

  • 植物細胞生物学

    2007
    -
    2011
    Institution name:新潟大学

  • 植物生理学I

    2007
    Institution name:新潟大学

▶ display all