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2005 Fiscal Year Final Research Report Summary

Mechanism of phytochrome signaling mediated by protein interaction

Research Project

Project/Area Number 16580076
Research Category

Grant-in-Aid for Scientific Research (C)

Allocation TypeSingle-year Grants
Section一般
Research Field Applied biochemistry
Research InstitutionKyoto University

Principal Investigator

KOHCHI Takayuki  Kyoto University, Graduate School of Biostudies, Professor, 生命科学研究科, 教授 (40202056)

Project Period (FY) 2004 – 2005
Keywordsphotoreceptor / phytochrome / chromophore / photomorphogenesis / Arabidopsis thaliana / protein interaction / phytochromobilin
Research Abstract

Sessile and photoautotrophic plants have developed very sensitive and sophisticated system to recognize light conditions properly for their survival and successful propagation. Phytochrome is a major photoreceptor in plants. However, the mechanisms of signal transduction mediated by phytochrome is largely unknown. The protein interaction should be a key mechanism for signaling for phytochrome since phytochrome have very specific character showing photoreversible conformation changes by illumination. To study protein interaction with phytochrome, the photoactive phytochrome with chromophore is required. By co-expression of heme oxygenase and various bilin reductase(s) in a single operon in conjunction with apophytochrome using two compatible plasmids, we developed a system to produce phytochromes with various chromophores in Escherichia coli. The metabolic engineering of bacteria for the production of various bilins for assembly into phytochromes will facilitate the molecular analysis of photoreceptors. We also generated transgenic plants that express epitope-tagged phytochromes. The introduced fusion genes were active and functional to rescue phytochrome mutant phenotype of phyA and phyB. Another approach we took to identify interacting proteins was yeast two-hybrid assays. A novel transcription factor containing zinc finger domain was identified. The specificity of interaction for wavelength was verified by using expressed proteins in vitro. The identified protein is a good candidate to mediated photobiological response since the knock-out mutant showed delayed flowering phenotype.
In addition to Arabidopsis works, we isolated several genes and corresponding mutants for heme oxygenase and phytochromobilin synthase for phytocrome chromophore biosynthesis from maize, pea and tomato. In particular, the tomato AUREA protein was expressed at higher levels in soluble form in E.coli.

  • Research Products

    (10 results)

All 2006 2005 2004

All Journal Article (10 results)

  • [Journal Article] Metabolic engineering to produce phytochromes with phytochromobilin, phycocyanobilin, or phycoerythrobilin chromophore in Escherichia coli.2006

    • Author(s)
      Mukougawa, K., Kanamotoa, H., Kobayashi, T., Yokota, A., Kohchi, T.
    • Journal Title

      FEBS Letters 580

      Pages: 1333-1338

    • Description
      「研究成果報告書概要(和文)」より
  • [Journal Article] Reconstitution of blue/green reversible photoconversion of a cyanobacterial photoreceptor, PixJl in phycocyanobilin producing E. coli.2006

    • Author(s)
      Yoshihara, S., Shimada, T., Matsuoka, D., Zikihara, K., Kohchi, T., Tokutomi, S.
    • Journal Title

      Biochemistry 45

      Pages: 3775-3781

    • Description
      「研究成果報告書概要(和文)」より
  • [Journal Article] Metabolic engineering to produce phytochromes with phytochromobilin, phycocyanobilin, or phycoerythrobilin chromophore in Escherichia coli.2006

    • Author(s)
      Mukougawa, K., Kanamotoa, H., Kobayashi, T., Yokota, A., Kohchi, T.
    • Journal Title

      FEBS Lett. 580

      Pages: 1333-1338

    • Description
      「研究成果報告書概要(欧文)」より
  • [Journal Article] Reconstitution of blue/green reversible photoconversion of a cyanobacterial photoreceptor. PixJ1 in phycocyanobilin producing E.coli.2006

    • Author(s)
      Yoshihara, S., Shimada, T., Matsuoka, D., Zikihara, K., Kohchi, T., Tokutomi, S.
    • Journal Title

      Biochem. 45

      Pages: 3775-3784

    • Description
      「研究成果報告書概要(欧文)」より
  • [Journal Article] Biosynthesis of chromophores for phytochrome and related photoreceptors in photosynthetic organisms2005

    • Author(s)
      Kohchi, T., Kataoka, H., Linley, P.J.
    • Journal Title

      Plant Biotechnology 22

      Pages: 409-413

    • Description
      「研究成果報告書概要(和文)」より
  • [Journal Article] The tomato photomorphogenetic mutant, aurea, is deficient in phytochromobilin synthase for phytochrome chromophore biosynthesis.2005

    • Author(s)
      Muramoto, T., Kami, C., Kataoka, H., Iwata, N., Linley, P.J., Mukougawa, K., Yokota, A., Kohchi, T.
    • Journal Title

      Plant Cell Physiology 46

      Pages: 409-413

    • Description
      「研究成果報告書概要(和文)」より
  • [Journal Article] The tomato photomorphogenetic mutant, aurea, is deficient in phytocbromobilin synthase for phytochrome chromophore biosynthesis.2005

    • Author(s)
      Muramoto, T., Kami, C., Kataoka, H., Iwata, N., Linley, P.J., Mukougawa, K., Yokota, A., Kohchi, T.
    • Journal Title

      Plant Cell Physiol. 46

      Pages: 661-665

    • Description
      「研究成果報告書概要(欧文)」より
  • [Journal Article] Biosynthesis of chromophores for phytoclirome and related photoreceptors in photosynthetic organisms.2005

    • Author(s)
      Kohchi, T., Kataoka, H., Linley, P.J.
    • Journal Title

      Plant Biotech. 22

      Pages: 409-413

    • Description
      「研究成果報告書概要(欧文)」より
  • [Journal Article] The Elml (ZmHy2) Gene of Maize Encodes a Phytochromobilin Synthase.2004

    • Author(s)
      Sawers, R.J., Linley, P.J., Gutierrez-Marcos, J.F., Delli-Bovi, T., Farmer, P.R., Kohchi, T., Terry, M.J., Brutnell, T.
    • Journal Title

      Plant Physiology 136

      Pages: 2771-2781

    • Description
      「研究成果報告書概要(和文)」より
  • [Journal Article] The Elm1 (ZmHy2) Gene of Maize Encodes a Phytochromobilin Synthase.2004

    • Author(s)
      Sawers, R.J., Linley, P.J., Gutierrez-Marcos, J.F., Delli-Bovi, T., Farmer, P.R., Kohchi, T., Terry M.J., Brutnell, T.P.
    • Journal Title

      Plant Physiol. 136

      Pages: 2771-2781

    • Description
      「研究成果報告書概要(欧文)」より

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Published: 2007-12-13  

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