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Study on mechanism of posttranslational regulation of ACC synthase by phosphorylation that involved in tomato fruit ripening

Research Project

Project/Area Number 17380020
Research Category

Grant-in-Aid for Scientific Research (B)

Allocation TypeSingle-year Grants
Section一般
Research Field Horticulture/Landscape architecture
Research InstitutionNagoya university

Principal Investigator

MORI Hitoshi  Nagoya Univ., Bioagricultural Sciences, Professor, 大学院生命農学研究科, 教授 (20220014)

Project Period (FY) 2005 – 2006
Project Status Completed (Fiscal Year 2006)
Budget Amount *help
¥15,800,000 (Direct Cost: ¥15,800,000)
Fiscal Year 2006: ¥4,500,000 (Direct Cost: ¥4,500,000)
Fiscal Year 2005: ¥11,300,000 (Direct Cost: ¥11,300,000)
Keywordsethylene biosynthesis / ACC synthase / turnover / protein phosphorylation / protein dephosphorylation / posttranslational regulation / タンパク質のリン酸化制御 / タンパク質脱リン酸化酵素
Research Abstract

ACC synthase (ACS) is a rate-limiting enzyme of ethylene biosynthesis pathway and each ACS isozyme expresses in response to various each stimulus. Recent studies suggested that ACS was regulated not only transcriptionally but also post-translationally. We found that LeACS2, a wound-inducible ACS in tomato (Lycopersicon esculentum Mill.), is immediately phosphorylated after translation at Ser-460 in the C-terminal region by calcium-dependent protein kinase (CDPK), and then acts in the phosphorylated form in the cell. Moreover, the results of treatments of kinase and phosphatase inhibitors showed that the half-life of phosphorylated LeACS2 was longer than that of non-phosphorylated LeACS2. These results suggest that phosphorylation/dephosphorylation regulates the turnover of LeACS2 protein in the cell and that dephosphorylation of LeACS2 causes degradation. Analyses of ethylene-overproducer (eto) mutants also supported our speculation. Thus, we attempted to identify the protein phosphatase involved in LeACS2 turnover. Biotin-tagged phospho-peptide (Biotinyl-KNNLRL(pS)FSKRMYD-CHO) based on the sequence in the neighborhood of Ser-460 of LeACS2 was synthesized. This peptide was incubated with the extract of wounded tomato fruit tissue. Proteins bound to the peptide were captured by streptavidin-bound Dynabeads and eluted by SDS. The eluate was subjected to SDS-PAGE and blotting, and detected with HRP conjugated streptavidin. As the results, 32 kDa protein was detected. The amount of the 32 kDa protein was dependent on the concentration of the phospho-peptide that was added to reaction mixture. Based on the molecular mass, we speculate the 32 kDa protein is a catalytic subunit of Ser/Thr protein phosphatase that is involved in dephosphorylation of LeACS2.

Report

(3 results)
  • 2006 Annual Research Report   Final Research Report Summary
  • 2005 Annual Research Report
  • Research Products

    (7 results)

All 2007 2006 2005

All Journal Article (6 results) Book (1 results)

  • [Journal Article] Subcellular localization and membrane topology of melon ethylene receptor CmERS1.2006

    • Author(s)
      Ma, B.
    • Journal Title

      Plant Physiology 141

      Pages: 587-597

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2006 Annual Research Report 2006 Final Research Report Summary
  • [Journal Article] Subcellular localization and membrane topology of melon ethylene receptor CmERS1.2006

    • Author(s)
      Ma, B.et al.
    • Journal Title

      Plant Physiology 141

      Pages: 587-597

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2006 Final Research Report Summary
  • [Journal Article] Auxin controls local cytokinin biosynthesis in the nodal stem in apical dominance.2006

    • Author(s)
      Tanaka, M.
    • Journal Title

      Plant Journal 45

      Pages: 1028-1036

    • Related Report
      2005 Annual Research Report
  • [Journal Article] Isolation, characterization and cloning of α-L-arabinofuranosidase expressed during fruit ripening of Japanese pear (Pyrus pyrifolia).2005

    • Author(s)
      Tateishi, A.
    • Journal Title

      Plant Physiology 138

      Pages: 1653-1664

    • Related Report
      2005 Annual Research Report
  • [Journal Article] An S-like ribonuclease gene is used to generate a trap-leaf enzyme in the carnivorous plant Drosera adelae.2005

    • Author(s)
      Okabe, T.
    • Journal Title

      FEBS Letters 579

      Pages: 5729-5733

    • Related Report
      2005 Annual Research Report
  • [Journal Article] Involvement of PPS3 phosphorylated by elicitor-responsive mitogen-activated protein kinases in the regulation of plant cell death.2005

    • Author(s)
      Katou, S.
    • Journal Title

      Plant Physiology 139

      Pages: 1914-1926

    • Related Report
      2005 Annual Research Report
  • [Book] 園芸生理学 分子生物学とバイオテクノロジー2007

    • Author(s)
      山木昭平 編
    • Total Pages
      314
    • Publisher
      文永堂出版
    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2006 Annual Research Report 2006 Final Research Report Summary

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Published: 2005-04-01   Modified: 2016-04-21  

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