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種子植物の生殖に関わるRNA結合タンパク質PPRの分子進化と機能の解明

Research Project

Project/Area Number 11J00134
Research Category

Grant-in-Aid for JSPS Fellows

Allocation TypeSingle-year Grants
Section国内
Research Field Breeding science
Research InstitutionKyoto University

Principal Investigator

藤井 壮太  京都大学, 大学院理学研究科, 特別研究員(SPD)

Project Period (FY) 2011 – 2013
Project Status Completed (Fiscal Year 2013)
Budget Amount *help
¥7,500,000 (Direct Cost: ¥7,500,000)
Fiscal Year 2013: ¥2,500,000 (Direct Cost: ¥2,500,000)
Fiscal Year 2012: ¥2,500,000 (Direct Cost: ¥2,500,000)
Fiscal Year 2011: ¥2,500,000 (Direct Cost: ¥2,500,000)
Keywordsミトコンドリア / 共進化 / RNA代謝 / RNA制御
Research Abstract

本研究では35アミノ酸の縦列繰り返しモチーフであるpentatricopeptide repeat (PPR)タンパク質がRNAを認識するメカニズム, およびPPRファミリーの種子植物の生殖進化への関わりを明らかにすることを目的としている. 本研究ではPPRのうち, Rf-PPRと呼ばれるサブグループに着目した. Rf-PPRはPPRの中でも特に多様性が高く, ミトコンドリアのRNA配列に対応して進化してきている. アブラナ科のモデル植物シロイヌナズナを用いる事で, Rf-PPRの機能解析, 分子進化解析を行い, PPRとミトコンドリアゲノムの共進化の理解を目的とした.
平成25年度は, Rf-PPRのうち, シロイヌナズナRFL2のノックアウト系統を得て機能解析を行った. トランスクプトーム解析を用いて, rf12変異体ではミトコンドリアのorf291という遺伝子由来のRNAのみが異常に蓄積することを明らかにした. 転写末端解析により, RFL2はorf291のRNAを内部で切断する機構に関与していることが示された. RNA ge1-shift assayとPPRがRNAに結合する法則性より, RFL2が高い特異性を持っと思われるorf291上の15塩基のRNA配列を明らかにした. この配列はRNA切断サイトより上流約70塩基の位置にあったため, 直接切断を行う酵素は他に存在することが示唆された. 近年, 植物ではタンパク質性のRNase P (PRORP)が発見されており, この酵素が切断活性を持っている可能性が考えられた. 従ってミトコンドリア局在のPRORPのノックダウンを行ったところ, 予想通りorf291の切断活性が抑制された. これはアダプターとなるPPRが制限酵素PRORPをRNA切断サイトに呼び込んでいる機構を示唆しており, 二分子によるRNA代謝メカニズムの存在を明らかにした.

Current Status of Research Progress
Current Status of Research Progress

1: Research has progressed more than it was originally planned.

Reason

申請書に記載したPPRコードの解読は共同研究によって第一段階を達成することができた. さらに, RFL2が結合するRNA配列の最小単位を決定できただけに留まらず, 協調的に機能する可能性があるRNA切断酵素も見出す事ができた.

Strategy for Future Research Activity

(抄録なし)

Report

(3 results)
  • 2013 Annual Research Report
  • 2012 Annual Research Report
  • 2011 Annual Research Report
  • Research Products

    (19 results)

All 2013 2012 2011

All Journal Article (8 results) (of which Peer Reviewed: 8 results) Presentation (10 results) (of which Invited: 2 results) Book (1 results)

  • [Journal Article] Mutagenesis of Individual Pentatricopeptide Repeat Motifs Affects RNA Binding Activity and Reveals Functional Partitioning of Arabidopsis PROTONGRADIENT REGULATION32013

    • Author(s)
      Fujii S, Sato N, Shikanai T
    • Journal Title

      Plant Cell

      Volume: 25 Issue: 8 Pages: 1-11

    • DOI

      10.1105/tpc.113.112193

    • Related Report
      2013 Annual Research Report
    • Peer Reviewed
  • [Journal Article] Function of PPR proteins in plastid gene expression2013

    • Author(s)
      Shikanai T, Fujii S
    • Journal Title

      RNA Biology

      Volume: 10 Issue: 9 Pages: 1-11

    • DOI

      10.4161/rna.25207

    • Related Report
      2013 Annual Research Report
    • Peer Reviewed
  • [Journal Article] The E domains of pentatricopeptide repeat proteins from different organelles are not functionally equivalent for RNA editing.2013

    • Author(s)
      Chateigner-Boutin AL, Colas des francs-Small C, Fujii S. Okuda K, Tanz SK, Small I.
    • Journal Title

      Plant J.

      Volume: (印刷中) Issue: 6 Pages: 935-945

    • DOI

      10.1111/tpj.12180

    • Related Report
      2012 Annual Research Report
    • Peer Reviewed
  • [Journal Article] Rice MPR25 encodes a pentatricopeptide repeat protein and is essential for RNA editing of nad5 transcripts in mitochondria.2012

    • Author(s)
      戸田拓士
    • Journal Title

      The Plant Journal

      Volume: 72 Issue: 3 Pages: 450-460

    • DOI

      10.1111/j.1365-313x.2012.05091.x

    • Related Report
      2012 Annual Research Report
    • Peer Reviewed
  • [Journal Article] A combinatorial amino acid code for RNA recognition by pentatricopeptide repeat proteins.2012

    • Author(s)
      Barkan A, Rojas M, Fujii S, Yap A, Chong YS, Bond CS, Suall I.
    • Journal Title

      PLoS Genet

      Volume: 8 Issue: 8 Pages: e1002910-e1002910

    • DOI

      10.1371/journal.pgen.1002910

    • Related Report
      2012 Annual Research Report
    • Peer Reviewed
  • [Journal Article] Rampant gene loss in the underground orchid Rhizanthella gardneri highlights evolutionary constraints on plastid genomes2011

    • Author(s)
      Delannoy E, Fujii S, Colas des Francs-Small C, Brundrett M, Small
    • Journal Title

      Molecular Biology and Evolution

      Volume: 28 Issue: 7 Pages: 2077-2086

    • DOI

      10.1093/molbev/msr028

    • Related Report
      2011 Annual Research Report
    • Peer Reviewed
  • [Journal Article] The evolution of RNA editing and pentatricopeptide repeat genes2011

    • Author(s)
      Fujii S, Small I
    • Journal Title

      New Phytologist

      Volume: 191 Issue: 1 Pages: 37-47

    • DOI

      10.1111/j.1469-8137.2011.03746.x

    • Related Report
      2011 Annual Research Report
    • Peer Reviewed
  • [Journal Article] Transcriptome map of plant mitochondria reveals islands of unexpected transcribed regions2011

    • Author(s)
      Fujii S, Toda T, Kikuchi S, Suzuki R, Yokoyama K, Tsuchida H, Yano K
    • Journal Title

      BMC Genomics

      Volume: 12 Issue: 1 Pages: 279-279

    • DOI

      10.1186/1471-2164-12-279

    • Related Report
      2011 Annual Research Report
    • Peer Reviewed
  • [Presentation] Member of rapidly evolving PPR subfamily cleaves RNA transcribed from active mitochondrial rearrangement site2013

    • Author(s)
      藤井 壮太
    • Organizer
      International Conference of Plant Mitochondrial Biology
    • Place of Presentation
      Ros Tower Hotel(ロサリオ, アルゼンチン)
    • Year and Date
      2013-05-12
    • Related Report
      2013 Annual Research Report
    • Invited
  • [Presentation] 巨大RNA結合タンパク質ファミリーによる転写後制御システム2013

    • Author(s)
      藤井 壮太
    • Organizer
      日本植物生理学会
    • Place of Presentation
      富山大学(富山県)
    • Year and Date
      2013-03-17
    • Related Report
      2013 Annual Research Report
    • Invited
  • [Presentation] RNA転写後制御システムの進化から見た細胞内共生の形2012

    • Author(s)
      藤井壮太
    • Organizer
      日本育種学会
    • Place of Presentation
      京都産業大学(京都府)(招待講演)
    • Year and Date
      2012-09-14
    • Related Report
      2012 Annual Research Report
  • [Presentation] オルガネラ進化の色即是空と育種2012

    • Author(s)
      藤井壮太
    • Organizer
      日本育種学会第121回講演会
    • Place of Presentation
      宇都宮大学(宇都宮市)(招待講演)
    • Year and Date
      2012-03-31
    • Related Report
      2011 Annual Research Report
  • [Presentation] Pentatr icopeptide repeatタンパク質PGR3の機能分化2012

    • Author(s)
      藤井壮太
    • Organizer
      日本植物生理学会
    • Place of Presentation
      岡山大学(岡山県)
    • Year and Date
      2012-03-22
    • Related Report
      2012 Annual Research Report
  • [Presentation] Rapid evolution of pentatricopeptide repeat genes involved in organelle gene2012

    • Author(s)
      Sota Fujii
    • Organizer
      第53回日本植物生理学会
    • Place of Presentation
      京都産業大学(京都市)
    • Year and Date
      2012-03-16
    • Related Report
      2011 Annual Research Report
  • [Presentation] Evolution of pentatricopeptide repeat gene reveals genetic conflict between nucleus and mitochondria in plant reproduction2012

    • Author(s)
      Sota Fujii
    • Organizer
      Plant Reproduction for Food 2012
    • Place of Presentation
      メルボルン(オーストラリア)(招待講演)
    • Year and Date
      2012-02-16
    • Related Report
      2011 Annual Research Report
  • [Presentation] 種子植物におけるpentatricopeptide repeat遺伝子ファミリーの進化2011

    • Author(s)
      藤井壮太
    • Organizer
      日本遺伝学会第83回大会
    • Place of Presentation
      京都大学(京都市)
    • Year and Date
      2011-09-20
    • Related Report
      2011 Annual Research Report
  • [Presentation] Diversifying selection of pentatricopeptide repeat genes reveal a conflict between nuclear and mitochondrial genomes throughout angiosperm evolution2011

    • Author(s)
      Sota Fujii
    • Organizer
      Society for Molecular Biology and Evolution 2011
    • Place of Presentation
      京都大学(京都市)
    • Year and Date
      2011-07-26
    • Related Report
      2011 Annual Research Report
  • [Presentation] Diversifying selection of restorer-like genes reveal a conflict between nuclear and mitochondrial genomes throughout angiosperm history2011

    • Author(s)
      Sota Fujii
    • Organizer
      International Conference for Plant Mitochondrial Biology 2011
    • Place of Presentation
      ホーエンローダ(ドイツ)(招待講演)
    • Year and Date
      2011-05-13
    • Related Report
      2011 Annual Research Report
  • [Book] Advances in Botanical Research, Mitochondrial genome evolution2013

    • Author(s)
      Budar F, Fujii S
    • Total Pages
      446
    • Publisher
      Elsevier
    • Related Report
      2013 Annual Research Report

URL: 

Published: 2011-12-12   Modified: 2024-03-26  

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