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種子貯蔵タンパク質の集積機構の解析

Research Project

Project/Area Number 14740438
Research Category

Grant-in-Aid for Young Scientists (B)

Allocation TypeSingle-year Grants
Research Field 植物生理
Research InstitutionKyoto University

Principal Investigator

嶋田 知生  京都大学, 大学院・理学研究科, 助手 (20281587)

Project Period (FY) 2002 – 2004
Project Status Completed (Fiscal Year 2004)
Budget Amount *help
¥4,000,000 (Direct Cost: ¥4,000,000)
Fiscal Year 2004: ¥900,000 (Direct Cost: ¥900,000)
Fiscal Year 2003: ¥1,300,000 (Direct Cost: ¥1,300,000)
Fiscal Year 2002: ¥1,800,000 (Direct Cost: ¥1,800,000)
Keywords貯蔵タンパク質 / 種子 / 選別輸送レセプター / 液胞 / プロテインボディ / 細胞内輸送 / シロイヌナズナ / PAC小胞 / AtVSR / PV72 / 液胞プロセシング酵素 / VPE / プロテインボデイ / 液胞輸送レセプター / タンパク質蓄積型液胞
Research Abstract

植物の種子には発芽後の成長のために,大量の種子貯蔵タンパク質が液胞内に蓄積している.本研究では,貯蔵タンパク質が短期間に大量集積するメカニズムの解析を行った.貯蔵タンパク質はまず前駆体として小胞体で合成され,液胞に輸送された後にプロセシングされ成熟型として蓄積する.シロイヌナズナ変異体を用いた解析で液胞プロセシング酵素(VPE)が実際に貯蔵タンパク質の成熟化に関っていることをin plantaで初めて証明した.VPEを欠損する変異体では貯蔵タンパク質の成熟化が完全に阻害されていた.また,同様にシロイヌナズナ変異体を用いて,液胞タンパク質の輸送に機能する選別輸送レセプター(VSR)を初めて特定した.AtVSR1欠損シロイヌナズナでは貯蔵タンパク質が前駆体として蓄積し,細胞外へ分泌されていた.さらに,貯蔵タンパク質の輸送に異常を示すシロイヌナズナ変異体を単離し解析した.免疫電子顕微鏡観察の結果,貯蔵タンパク質を細胞外に分泌する系統と,分泌せず細胞内に異常な構造物が形成する系統が見つかった.前者はAtVSR1変異体と同様に貯蔵タンパク質の液胞への選別過程に欠損があると考えられた.後者の構造物は小胞体に由来すると考えられることから,輸送過程の初期に欠損があると考えられた.一部の変異体については原因遺伝子の特定を行い,分裂酵母の液胞タンパク質輸送に関る因子と相同性が見られた。これらの解析により,貯蔵タンパク質の生合成に関わる中心的な因子とともに,その全体像についても明らかにすることができた.

Report

(3 results)
  • 2004 Annual Research Report
  • 2003 Annual Research Report
  • 2002 Annual Research Report
  • Research Products

    (14 results)

All 2005 2004 Other

All Journal Article (3 results) Publications (11 results)

  • [Journal Article] Endosomal proteases facilitate the fusion of endosomes with vacuoles at the final step of the endocytotic pathway.2005

    • Author(s)
      Kenji Yamada
    • Journal Title

      Plant Journal 41(6)

      Pages: 888-898

    • Related Report
      2004 Annual Research Report
  • [Journal Article] Diversity and formation of endoplasmic reticulum-derived compartments in plants. Are these compartments specific to plant cells?2004

    • Author(s)
      Ikuko Hara-Nishimura
    • Journal Title

      Plant Physiology 136(3)

      Pages: 3435-3439

    • Related Report
      2004 Annual Research Report
  • [Journal Article] Endoplasmic reticulum-resident proteins are constitutively transported to vacuoles for degradation.2004

    • Author(s)
      Kentaro Tamura
    • Journal Title

      Plant Journal 39(3)

      Pages: 393-402

    • Related Report
      2004 Annual Research Report
  • [Publications] Etsuko Watanabe: "An ER-Localized Form of PV72, a Seed-Specific Vacuolar Sorting Receptor, Interferes the Transport of an NPIR-Containing Proteinase in Arabidopsis Leaves."Plant Cell Physiology. 45・1. 9-17 (2004)

    • Related Report
      2003 Annual Research Report
  • [Publications] Tomoo Shimada: "Vacuolar sorting receptor for seed storage proteins in Arabidopsis thaliana."Proc.Natl.Acad.Sci.USA.. 100・26. 16095-16100 (2003)

    • Related Report
      2003 Annual Research Report
  • [Publications] Takashi Okamoto: "C-terminal KDEL sequence of a KDEL- tailed cysteine proteinase (sulfhydryl-endopeptidase) is involved in formation of KDEL vesicle and in efficient vacuolar transport of sulfhydryl-endopeptidase."Plant Physiology. 132・4. 1892-1900 (2003)

    • Related Report
      2003 Annual Research Report
  • [Publications] Kentaro Tamura: "Why green fluorescent fusion proteins have not been observed in the vacuoles of higher plants."Plant J.. 35・4. 545-555 (2003)

    • Related Report
      2003 Annual Research Report
  • [Publications] Ryo Matsushima: "The ER body, a novel endoplasmic reticulum-derived structure in Arabidopsis."Plant Cell Physiology. 44・7. 661-666 (2003)

    • Related Report
      2003 Annual Research Report
  • [Publications] Tomoo Shimada: "Vacuolar processing enzymes are essential for proper processing of seed storage proteins in Arabidopsis thaliana."J Biol.Chem.. 278・34. 32292-32299 (2003)

    • Related Report
      2003 Annual Research Report
  • [Publications] 嶋田 知生: "細胞工学「VPE/legumainによる液胞・リソソームタンパク質の成熟化」"秀潤杜. 2 (2003)

    • Related Report
      2003 Annual Research Report
  • [Publications] Shimada: "A vacuolar sorting receptor PV72 on the membrane of vesicle that accumulate precursors of seed storage proteins (PAC vesicles)"Plant and Cell Physiology. 43・10. 1086-1095 (2002)

    • Related Report
      2002 Annual Research Report
  • [Publications] Matsushima: "An endoplasmic reticulum-derived structure that is induced under stress conditions in Arabidopsis"Plant Physiology. 130. 1807-1814 (2002)

    • Related Report
      2002 Annual Research Report
  • [Publications] Watanabe: "Calcium-mediated association of a putative vacuolar sorting receptor PV72 with a prpeptide of 2S albumin"J. Biological Chemistry. 277. 8708-8715 (2002)

    • Related Report
      2002 Annual Research Report
  • [Publications] 嶋田知生, 西村いくこ: "朝倉植物生理学講座 1 植物細胞「液胞」"朝倉書店・西村幹夫編. 10 (2002)

    • Related Report
      2002 Annual Research Report

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Published: 2002-04-01   Modified: 2016-04-21  

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