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Regulation of gene expression by visible light

Research Project

Project/Area Number 04454610
Research Category

Grant-in-Aid for General Scientific Research (B)

Allocation TypeSingle-year Grants
Research Field 分子遺伝学・分子生理学
Research InstitutionKYOTO UNIVERSITY

Principal Investigator

INOKUCHI Hachiro  Faculty of Science, Department of Biophysics, Associate Professor, 理学部, 助教授 (20028195)

Project Period (FY) 1992 – 1993
Project Status Completed (Fiscal Year 1993)
Budget Amount *help
¥6,200,000 (Direct Cost: ¥6,200,000)
Fiscal Year 1993: ¥2,400,000 (Direct Cost: ¥2,400,000)
Fiscal Year 1992: ¥3,800,000 (Direct Cost: ¥3,800,000)
Keywordslight-sensitive mutant of E.coli / biosynthesis of heme / biosynthesis of chlorophyll / hemK / protoporphyrin IX / active species of oxygen / ferrochelatase / cDNA clones of the hemA and hemH from barley and cucumber / 光感受性変異株 / ΔvisA(=ΔhemH) / hemE / hemG / hemK / 大腸菌 / 可視光感受性 / ヘム合成系 / visA遺伝子 / visB遺伝子 / ユビキノン生合成系
Research Abstract

Some intermediates in the biosynthetic pathway generate an active pecies of oxygen via photochemical reactions upon illumination by visible light. For example, it is known that protoporphyrin IX, a intermediate in the biosynthesis of heme, is such a photosensitizer. Recently, Several light-sensitive mutant of E.coli have been isolated and characterized in my laboratory. The cells with mutantions in the visA (hemH) gene encoding ferrochelatase, the enzyme that catalyzes the final step in the biosynthesis of heme, are killed by illumination with visible light. The photoresistant mutants from the visA (hemH)-deleted strain have been isolated. Since on true revertant of visA (hemH) were expected, the revertats should be double mutants in the visA gene and in related genes that is involved in the heme biosynthetic pathway at a step before the reaction catayzed by ferrochelatase. In this research project, the studies on the genes involved in the biosynethsis of heme have been done in connection with light-responses of the cells.
(1) Using mutants isolated, the hemE and hemG gene of E.coli were cloned and sequenced.
(2) Two new genes involved in the biosynthesis of heme were identified. A new gene, designated hemK, composes a hemA-prfA-hemK operon at 27 min on the likage map. The hemK encodes 225 amino acids protein. Because the mutant cells accumulate protoporphyrinogen, the defect may locate in a gene functionally equvalent with the hemG gene.
(3) The cDNA clones of the hemA and hemH from barley and cucumber were isolated by a method of complementation with the visA-deleted or the hemA-deleted mutant of E.coli.
(4) A high expression system of the enzyme ferrochelatase was constructed and a large amount of the enzyme were prepared. The physical and biochemical natures of the purified enzyme were investigated. The crystallographic studies of this enzyme are under way.

Report

(3 results)
  • 1993 Annual Research Report   Final Research Report Summary
  • 1992 Annual Research Report
  • Research Products

    (22 results)

All Other

All Publications (22 results)

  • [Publications] K.Nishimura: "Cloning and Sequencing of the hemE gene encoding uroporphyrinogen III decarboxylase of E.coliK-12" Gene. 133. 109-113 (1993)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1993 Final Research Report Summary
  • [Publications] M.Kitabatake: "A simplified method for generating stepwise deletions using PCR" Gene. 123. 59-61 (1993)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1993 Final Research Report Summary
  • [Publications] K.Nishimura: "Location of the ubiA gene on the physical map of Escherichia coli" J.Bacteriol.174. 5762 (1992)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1993 Final Research Report Summary
  • [Publications] K.Nakahigashi: "Isolation and characterization of a light-sensitive mutant of E.coliK12 with a mutation in a gene that is required for the biosynthesis of Ubiquinone" J.Bacteriol.174. 7352-7359 (1992)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1993 Final Research Report Summary
  • [Publications] T.Masuda: "Sequence and expression of tRNA^<Glu> gene of cucumber chloroplast genome" Plant Physiol.Biochem.30. 235-243 (1992)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1993 Final Research Report Summary
  • [Publications] K.Miyamoto: "Accumulation of protoporphyrin IX in light-sensitive mutants of Escherichia coli" FEBS Letters. 310. 246-248 (1992)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1993 Final Research Report Summary
  • [Publications] K.Nishimura et al.: "Cloning and Sequencing of the hemE gene encoding uroporphyrinogen III decarboxylase of E.coli K12" Gene. 133. 109-113 (1993)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1993 Final Research Report Summary
  • [Publications] M.Kitabatake & H.Inokuchi: "A Simplefied method for generating step-wise deletions using PCR" Gene. 123. 57-61 (1993)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1993 Final Research Report Summary
  • [Publications] M.Nishimura et al.: "Location of the ubiA gene on the physical map of Escherichia coli" J.Bacteriol. 174. 5762 (1992)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1993 Final Research Report Summary
  • [Publications] K.Nakahigashi et al.: "Isolatioon and characterization of a light-sensitive mutant of E.coli K12 with a mutation in a gene that is required for the biosynthesis of ubiquinone" J.Bacteriol. 174. 7352-7359 (1992)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1993 Final Research Report Summary
  • [Publications] T.Musuda et al.: "Sequence and expression of tRNA^<Glu> gene of cucumber chloroplast genome" Plant Physiol.Biochem. 30. 235-243 (1992)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1993 Final Research Report Summary
  • [Publications] K.Miyamoto et al.: "Accumlation of protoporphyrin IX in light-sensitive mutants of Escherichia coli" FEBS. 310. 236-248 (1992)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1993 Final Research Report Summary
  • [Publications] K.Nishimura: "Location of the ubiA gene on the physical map of Escherichia coli" J.Bacteriol. 174. 5762 (1992)

    • Related Report
      1993 Annual Research Report
  • [Publications] K.Nakahigashi: "Isolation and characterization of a light-sensitive mutant of E.coli K12 with a mutation in a gene that is required for the biosynthesis of ubiquinone" J.Bacteriol. 174. 7352-7359 (1992)

    • Related Report
      1993 Annual Research Report
  • [Publications] T.Masuda: "Sequence and expression of tRNA^<Glu>gene of cuccumber chloroplast genome" Plant Physiol.Biochem.30. 235-243 (1992)

    • Related Report
      1993 Annual Research Report
  • [Publications] K.Miyamoto: "Accumulation of protoporphyrin IX in light-Sensitive mutants of Escherichia coli" FEBS Letters. 310. 246-248 (1992)

    • Related Report
      1993 Annual Research Report
  • [Publications] K.Nishimura: "Cloning and Sequencing of the hemE gene encoding uroporphyrinogen III decarbexylase of E.coli K-12" Gene. 133. 109-113 (1993)

    • Related Report
      1993 Annual Research Report
  • [Publications] M.Kitabatake: "A simplified method for generating step-wise deletions using PCR" Gene. 123. 59-61 (1993)

    • Related Report
      1993 Annual Research Report
  • [Publications] K.Nakahigashi,K.Miyamoto,K.Nishimura & H.Inokuchi: "Isolation and Characterigation of a light-sensitive mutant of Escherichia Coli K-12 with a mutation in a gone that is rdquired for the biosynthesis of Uiquinono" Journal of Bacteriology. 174. 7352-7359 (1992)

    • Related Report
      1992 Annual Research Report
  • [Publications] K.Miyamoto,K.Nishimura,T.Masuda,H.Tsuji & H.Inokuchi: "Accumulation of piotoporphyrin IX in light-sensitive mutants of Escherichia coli" FEBS Letters. 310. 246-248 (1992)

    • Related Report
      1992 Annual Research Report
  • [Publications] K.Nishimura,T.Nakayashiki,& H.Inokuchi: "Cloning and sequencing of the hem E gene encoding uroporphyrinogen3 decarboxylase of Escherichia Coli K12"

    • Related Report
      1992 Annual Research Report
  • [Publications] H.Yang,H.Inokuchi & J.Adler: "Behavioral responses to light of an Escherichia Coli mutant accumulating Protoporphyrin IX"

    • Related Report
      1992 Annual Research Report

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Published: 1992-04-01   Modified: 2016-04-21  

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