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Functional design of enzymes for glutamate synthesis and factors involved in cooperative assimilation of carbon and nitrogen

Research Project

Project/Area Number 10640630
Research Category

Grant-in-Aid for Scientific Research (C)

Allocation TypeSingle-year Grants
Section一般
Research Field 植物生理
Research InstitutionOsaka University

Principal Investigator

HASE Toshiharu  Osaka University, Institute for Protein Research, Professor, たんぱく質研究所, 教授 (00127276)

Co-Investigator(Kenkyū-buntansha) ARIGA Yoko  Osaka University, Institute for Protein Research, Instructor, たんぱく質研究所, 助手 (60255429)
FUJITA Yuichi  Osaka University, Institute for Protein Research, Instructor, たんぱく質研究所, 助手 (80222264)
Project Period (FY) 1998 – 2000
Project Status Completed (Fiscal Year 2000)
Budget Amount *help
¥3,500,000 (Direct Cost: ¥3,500,000)
Fiscal Year 2000: ¥700,000 (Direct Cost: ¥700,000)
Fiscal Year 1999: ¥700,000 (Direct Cost: ¥700,000)
Fiscal Year 1998: ¥2,100,000 (Direct Cost: ¥2,100,000)
KeywordsCarbon assimilation / Nitrogen assimilation / Ferredoxin / Glutamate Synthase / Ferredoxin-NADP^+ reductase / Electron transfer complex / フェレドキシンNADP^+還元酵素 / 炭酸固定 / シアノバクテリア / ラン藻 / 炭素同化
Research Abstract

Assimilation of carbon and nitrogen in photosynthetic organisms is dependent on the supply of reductant and ATP and the regulation of the metabolic cross-talk of these metabolic processes is crucial for cell growth. This project focused on enzymes involved in glutamate synthesis, which is the first product of C and N assimilation, and reductant supply system for catalysis. Following results have been obtained.
1. Genes for Fd- and NADPH-dependent glutamate synthase were cloned from the cyanobacteria Plectonema boryanum. Mutants deficient of each gene is viable, but only the strain lacking Fddependent enzyme is suffered from nitrogen deficiency under an efficient carbon assimilation condition. In this strain, change of cellular amino acid pool, decrease in the transcript level of phycobiliporoteins and efficient reduction from nitrate to ammonia were observed in response to the nitrogen deficiency.
2. Mesophyll and bundle sheath cells of maize leaves, which are differentiated in C and N assimilation, possess unique ferredoxin isoproteins (FdI and FdII, respectively). Using cyanobacterial cells whose endogenous Fd gene was replaced by each maize Fd gene, it was found that FdI functions predominantly as an electron carrier for the non-cyclic photosynthetic electron transfer and that FdII was superior in the cyclic electron transfer, resulting in a weak electron donation to the N-assimilation pathway. These results suggest that Fd is a key molecule for the coordination of C and N assimilation.
3.X-ray crystal structure of the complex between Fd and Fd-NADP+ reductase has been determined. Structural information on topology of the prosthetic groups and interaction of the two proteins are obtained. To obtain structural basis of the cross-talk of C, N and S assimilation, this study will be extended to investigate molecular designs of Fd for interaction with GOGAT and other S assimilatory redox enzymes.

Report

(4 results)
  • 2000 Annual Research Report   Final Research Report Summary
  • 1999 Annual Research Report
  • 1998 Annual Research Report
  • Research Products

    (23 results)

All Other

All Publications (23 results)

  • [Publications] T.Matsumura: "cDNA cloning and characterization of ferredoxin localized in bundle-sheath cell of maize leaves"Plant Physiology. 199. 491-498 (1999)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] H.Okuhara: "Cloning and inactivation of genes encoding ferredoxin- and NADH-dependent glutamate synthase in the cyanobacterium Plectonema boryanum : imbalances in nitrogen and carbon assimilations caused by the deficiency of the ferredoxin-dependent enzyme."Plant Physiology. 120. 33-41 (1999)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] T.Akashi: "Comparison of the electrostatic binding sites on the surface of ferredoxin for two ferredoxin-dependent enzymes, ferredoxin-NADP^+oxidoreductase and sulfite reductase."Journal of Biological Chemistry. 274. 29399-29405 (1999)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] K.Youekura-Sakahi: "Analysis of reductant supply systems for ferredoxin dependent sulfite reductase in photosynthetic and nonphotosynthetic organs of maize."Plant Physiology. 122. 887-894 (2000)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] Y.Onda: "Differential interaction of maize root ferredoxin-NADP^+oxidoreductase with photosynthetic and nonphotosynthetic ferredoxin isoproteins."Plant Physiology. 123. 1037-1045 (2000)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] Y.Kimata-Ariga: "Differential electron flow around photosystem I by two C4-photosynthetic-cell-specific ferredoxins."EMBO Journal. 19. 5041-5050 (2000)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] M.Nakayama: "Plant sulfite reductase : molecular structure, catalytic function and interaction with ferredoxin."Journal of Inorganic Biochemistry. 82. 27-32 (2000)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] G.Kurisu: "Structure of the electron transfer complex between ferredoxin and ferredoxin-NADP^+ reductase."Nature Structural Biology. 8. 117-121 (2001)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] Matsumura T., Kimata-Ariga Y., Sakakibara H., Sugiyama T., Murata H., Takao T., Shimonishi Y.and Hase T.: "cDNA cloning and characterization of ferredoxin localized in bundle-sheath cell of maize leaves."Plant Physiol. 199. 491-498 (1999)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] Okuhara H., Matsumura T., Fujita Y.and Hase T.: "Cloning and inactivation of genes encoding ferredoxin- and NADH-dependent glutamate synthase in the cyanobacterium Plectonema boryanum : imbalances in nitrogen and carbon assimilations caused by the deficiency of the ferredoxin-dependent enzyme."Plant Physiol. 120. 33-41 (1999)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] Akashi T., Matsumura T., Ideguchi T, Iwakiri K., Kawakatsu T., Taniguchi I.and Hase T.: "Comparison of the electrostatic binding sites on the surface of ferredoxin for two ferredoxin-dependent enzymes, ferredoxin-NADP^+ oxidoreductase and sulfite reductase."J.Biol.Chem.. 274. 29399-29405 (1999)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] Yonekura-Sakakibara K., Onda Y., Ashikari T., Tanaka Y., Kusumi T.and Hase T.: "Analysis of reductant supply systems for ferredoxin dependent sulfite reductase in photosynthetic and nonphotosynthetic organs of maize."Plant Physiol.. 122. 887-894 (2000)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] Onda Y., Matsumura T., Kimata-Ariga Y., Sakakibara H., Sugiyama T.and Hase T.: "Differential interaction of maize root ferredoxin-NADP^+ oxidoreductase with photosynthetic and nonphotosynthetic ferredoxin isoproteins."Plant Physiol.. 123. 1037-1045 (2000)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] Kimata-Ariga Y., Matsumura T., Kada S., Fujimoto H., Fujita Y., Endo T., Mano J., Sato F.and Hase T.: "Differential electron flow around photosystem I by two C4-photosynthetic-cell-specific ferredoxins."EMBO J. 19. 5041-5050 (2000)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] Nakayama M., Akashi T.and Hase T.: "Plant sulfite reductase : molecular structure, catalytic function and interaction with ferredoxin."J.Inorg.Biochem.. 82. 27-32 (2000)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] Kurisu G., Kusunoki M., Katoh E., Yamazaki T., Teshima K., Onda Y., Kimata-Ariga Y.and Hase T.: "Structure of the electron transfer complex between ferredoxin and ferredoxin-NADP^+ reductase."Nature Struct.Biol.. 8. 117-121 (2001)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] Yayoi Onda: "Differential interaction of maige root ferredoxin-NADP^+ reductase with photasynthetic and nonphotosynthetic ferredoxin isoproteins"Plant Physiol.. 123. 1037-1045 (2000)

    • Related Report
      2000 Annual Research Report
  • [Publications] Yoko Kimata.Ariga: "Differential electron flow around photosystem I by two C_4-photosynthefic-cell-specific ferredoxins"EMBO J.. 19. 5041-5050 (2000)

    • Related Report
      2000 Annual Research Report
  • [Publications] Genji Kurisu: "Structure of the electron transfer complex between ferredoxin and ferredoxin-NADP^+ reductase"Nature Struct.Biol.. 8. 117-121 (2001)

    • Related Report
      2000 Annual Research Report
  • [Publications] Hiroaki Okuhara: "Cloning and inactivation of gens encoding ferredoxin-and NADPH-dependent glutamate synthase in the cyanobacterium Plectonema boryanum"Plant Physiology. Vol.120. 33-41 (1999)

    • Related Report
      1999 Annual Research Report
  • [Publications] Tetsuyuki Akashi: "Comparison of the electrostatic binding sites on the surface of ferredoxin for two ferredoxin-dependent enzyme, ferredoxin-NADP^+ reductase"The Journal of Biological Chemistry. Vol.274. 29399-29405 (1999)

    • Related Report
      1999 Annual Research Report
  • [Publications] Tomohiro Matsumura: "Complementary DNAcloning and characterigation of rerredoxin localiged in Bundle-sheath cells of maige leaves" Plant Physiology. 119. 481-488 (1999)

    • Related Report
      1998 Annual Research Report
  • [Publications] Hiroaki Okuhara: "cloning and inactivation of genes encoding ferredoxin-and NADH-dependent glutamate synthase in the cyanobocterium Plectonema boryanum" Plant Physiology. 120(in Press). (1999)

    • Related Report
      1998 Annual Research Report

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Published: 1998-04-01   Modified: 2016-04-21  

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