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Analysis of mRNP remodeling during NMD

Research Project

Project/Area Number 19770151
Research Category

Grant-in-Aid for Young Scientists (B)

Allocation TypeSingle-year Grants
Research Field Molecular biology
Research InstitutionYokohama City University

Principal Investigator

山下 暁朗  Yokohama City University, 医学研究科, 客員准教授 (20405020)

Project Period (FY) 2007 – 2008
Project Status Completed (Fiscal Year 2008)
Budget Amount *help
¥3,820,000 (Direct Cost: ¥3,400,000、Indirect Cost: ¥420,000)
Fiscal Year 2008: ¥1,820,000 (Direct Cost: ¥1,400,000、Indirect Cost: ¥420,000)
Fiscal Year 2007: ¥2,000,000 (Direct Cost: ¥2,000,000)
Keywords遺伝情報複製 / 転写装置 / 再編 / 制御
Research Abstract

本研究により、動物細胞内からの高精度mRNP複合体精製システムを新たに構築することに成功した。具体的には、mRNP精製のためのタグであるBoxB配列を挿入したbeta-globin遺伝子由来のmRNA(翻訳型mRNA)とその5'非翻訳領域に翻訳開始を阻害する強固なステムループを形成する配列を挿入した遺伝子由来のmRNA(非翻訳型mRNA)をmRNP精製アフィニティータグを用いて精製した。mRNP精製アフィニティータグとして、BoxB結合ペプチドであるバクテリオファージNタンパク質(Nペプチド)にStreptavidin binding peptide(SBP)とGSTとFlag epitope tagを融合した物を使用した。この解析により、動物細胞内で翻訳中のリボソームが結合したmRNP(リボソーム、翻訳伸張因子を含む、スプライシング依存的mRNA結合タンパク質複合体であるEJCを含まない)と翻訳されていないmRNP(EJCをふくむ、リボソーム翻訳伸張因子を含まない)の構成因子が大きく異なることを生化学的に明らかとした。この結果は、動物細胞内で翻訳依存的にmRNPの大きな再構成が起こるということを初めて示した物である。

Report

(3 results)
  • 2008 Annual Research Report   Final Research Report ( PDF )
  • 2007 Annual Research Report
  • Research Products

    (18 results)

All 2009 2008 2007

All Journal Article (8 results) (of which Peer Reviewed: 4 results) Presentation (10 results)

  • [Journal Article] SMG-8 and SMG-9, two novel subunits of the SMG-1 complex, regulate remodeling of the mRNA surveillance complex during nonsense-mediated mRNA decay.2009

    • Author(s)
      Yamashita A, Izumi N, Kashima I, Ohnishi T, Saari B, Katsuhata Y, Muramatsu R, Morita T, Iwamatsu A, Kurata R, Hachiya T, Hirano H, Anderson P, Ohno S
    • Journal Title

      Genes Dev 23(9)(in press)

    • Related Report
      2008 Final Research Report
  • [Journal Article] SMG-8 and SMG-9, two novel subunits of the SMG-1 complex, regulate remodeling ofthe mRNA surveillance complex during nonsense-mediated mRNA decay2009

    • Author(s)
      Akio Yamashita, Natsuko Izumi, Isao Kashima, Tetsuo Ohnishi, Bonnie Saari, Yukiko Katsuhata, Reiko Muramatsu, Tomoko Morita, Akihiro Iwamatsu, Rie Kurata, Takahisa Hachiya, Hisashi Hirano, Philip Anderson. and Shigeo Ohno
    • Journal Title

      Genes and development 23

    • Related Report
      2008 Annual Research Report
    • Peer Reviewed
  • [Journal Article] Versatile applications of transcriptional pulsing to study mRNA turnover in mammalian cells2007

    • Author(s)
      Chen CY, Yamashita Y, Chang TC, Yamashita A, Zhu W, Zhong Z, Shyu AB
    • Journal Title

      RNA (10)

      Pages: 1775-1786

    • Related Report
      2008 Final Research Report
  • [Journal Article] Human TOB, an antiproliferative transcription factor, is a poly(A)-binding protein-dependent positive regulator of cytoplasmic mRNA deadenylation.2007

    • Author(s)
      Ezzeddine N, Chang TC, Zhu W, Yamashita A, Chen CY, Zhong Z, Yamashita Y, Zheng D, Shyu AB
    • Journal Title

      Mol. Cell. Biol. 27(22)

      Pages: 7791-7801

    • Related Report
      2008 Final Research Report
  • [Journal Article] Distant N- and C-terminal domains are required for intrinsic kinase activity of SMG-1, a critical component of nonsense-mediated mRNA decay.2007

    • Author(s)
      Morita T, Yamashita A, Kashima I, Ogata K, Ishiura S, Ohno S.
    • Journal Title

      J Biol Chem. 16 ; 282(11)

      Pages: 7799-7808

    • Related Report
      2008 Final Research Report
  • [Journal Article] Human TOB,an antiproliferative transcription factor,is apoly(A)-binding protein-dependent positive regulator of cyto plasmic mRNA deadenylation.2007

    • Author(s)
      Ezzeddine N, Chang TC, Zhu W, Yamashita A, Chen CY, Zhong Z, Yamashita Y, Zheng D, Shyu AB
    • Journal Title

      Mol.Cell.Biol 27

      Pages: 7791-7801

    • Related Report
      2007 Annual Research Report
    • Peer Reviewed
  • [Journal Article] Versatile applications of transcriptional pulsing to study m RNA turnover in mammalian cells2007

    • Author(s)
      Chen CY, Yamashita Y, Chang TC, Yamashita A, Zhu W, Zhong Z, Shyu AB
    • Journal Title

      RNA 13

      Pages: 1775-1786

    • Related Report
      2007 Annual Research Report
    • Peer Reviewed
  • [Journal Article] Distant N-andC-terminal domatns are required fbr intrinsic kinase activity of SMG-1,a critical component of nonsense-mediated mRNA decay2007

    • Author(s)
      Morita T, Yamashita A, Kashima I, Ogata K, Ishiura S, Ohno S
    • Journal Title

      J.Biol., Chem 282

      Pages: 7799-7808

    • Related Report
      2007 Annual Research Report
    • Peer Reviewed
  • [Presentation] TRANSLATION TERMINATION COMPLEX DURING NONS ENSE- MEDIATED MRNA DECAY2008

    • Author(s)
      Yamashita A, Izumi N, Kashima I, Katsuhata Y, Muramatsu R. Ohno S
    • Organizer
      2008 Cold spring harver labatory meeting on translational control
    • Place of Presentation
      New York, USA
    • Year and Date
      2008-09-03
    • Related Report
      2008 Annual Research Report
  • [Presentation] The formation of SMG-1 : Upf1 : eRF1 : eRF3 complex (SURF) on ribosomes precedes translation dependent Upf1 phosphorylation during nonsense-mediated mRNA decay2008

    • Author(s)
      Yamashita A, Kashima I, Izumi N, Katsuhata Y, Muramatsu R, Ohno S
    • Organizer
      THE 21st NAITO CONFERENCE Nuclear Dynamics and RNA [I]
    • Place of Presentation
      Yamanashi
    • Year and Date
      2008-06-26
    • Related Report
      2008 Final Research Report
  • [Presentation] The formation of SMG-1 : Upfl : eRF1 : eRF3 complex (SURF) on ribosomes precedes translation-dependent-Upfl phosphorylatio n during nonsense-mediated mRNA decay2008

    • Author(s)
      Yamashita A, Kashima I, Izumi N, Katsuhata Y, Muramatsu R, Ohno S
    • Organizer
      THE 21st NAITO CONFERENCE "Nuclea Dynamics and RNA [I]
    • Place of Presentation
      Yamanashi, Japan
    • Year and Date
      2008-06-26
    • Related Report
      2008 Annual Research Report
  • [Presentation] TRANSLATION TERMINATION COMPLEX DURING NONSENSE- MEDIATED MRNA DECAY.2008

    • Author(s)
      Yamashita A, Izumi N, Kashima I, Katsuhata Y, Muramatsu R, Ohno S.
    • Organizer
      Cold spring harbor laboratory meeting 2008 Translational control
    • Place of Presentation
      New York
    • Related Report
      2008 Final Research Report
  • [Presentation] 翻訳依存的なmRNA監視複合体形成および再構成がUpf1リン酸化を誘導する2007

    • Author(s)
      山下 暁朗, 泉 奈津子, 鹿島 勲, 森田 智子, 村松 玲子
    • Organizer
      第30回日本分子生物学会年会〜第80回日本生化学会大会合同大会
    • Place of Presentation
      パシフィコ横浜
    • Year and Date
      2007-12-15
    • Related Report
      2007 Annual Research Report
  • [Presentation] The SMG-1 : SMG-8 : p60 Complex Regulated Phosphorylation Of Upf1 Is Required For The Dissociation Of eRFs From mRNA Surveillance Complex During Nonsense-mediated mRNA decay.2007

    • Author(s)
      Yamashita A, Izumi N, Kashima I, Ohnishi T, Anderson P, Ohno S
    • Organizer
      第9回日本RNA学会年会
    • Place of Presentation
      名古屋国際会議場
    • Year and Date
      2007-07-31
    • Related Report
      2008 Final Research Report
  • [Presentation] The SMG-1:SMG-8:p60 Complex Regulated Phosphorylation Of Upfl Is Required For The Dissociation Of eRFs From mRNA Surveillance Complex During Nonsense-mediated mRNA decay2007

    • Author(s)
      Yamashita A, Izumi N, Kashima I, Ohnishi T, Anderson P, Ohno S
    • Organizer
      第9回日本RNA学会年会
    • Place of Presentation
      名古屋国際会議場
    • Year and Date
      2007-07-31
    • Related Report
      2007 Annual Research Report
  • [Presentation] The SMG-1:SMG-8:p60 Complex Regulated Phosphorylation Of Upfl Is Required For The Dissociation Of eRFs From mRNA Surveillance Comp lex During Nonsense-mediated mRNA decay2007

    • Author(s)
      Yamashita A, Kashima I, Izumi N, Ohnishi T, Anderson P, Ohno S
    • Organizer
      第12回国際RNA学会年会
    • Place of Presentation
      University of Wisconsin,Madison
    • Year and Date
      2007-06-03
    • Related Report
      2007 Annual Research Report
  • [Presentation] 翻訳依存的なmRNA監視複合体形成および再構成がUpf1リン酸化を誘導する2007

    • Author(s)
      山下暁朗,泉奈津子,鹿島勲,森田智子,村松玲子,大野茂男
    • Organizer
      BMB2007-第30回日本分子生物学会年会・第80回日本生化学会大会合同大会
    • Place of Presentation
      横浜
    • Related Report
      2008 Final Research Report
  • [Presentation] The SMG-1 : SMG-8 : p60 Complex Regulated Phosphorylation Of Upf1 Is Required For The Dissociation Of eRFs From mRNA Surveillance Complex During Nonsense-mediated mRNA decay.2007

    • Author(s)
      Yamashita A, Kashima I, Izumi N, Ohnishi T, Anderson P, Ohno S.
    • Organizer
      第12回国際RNA学会年会
    • Place of Presentation
      Wisconsin
    • Related Report
      2008 Final Research Report

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Published: 2007-04-01   Modified: 2016-04-21  

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