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生殖細胞の形成を制御するNanos/Pumilio/mRNA複合体の構造解析

Publicly Offered Research

Project AreaStructural basis of cell-signalling complexes mediating signal perception, transduction and responses
Project/Area Number 23121524
Research Category

Grant-in-Aid for Scientific Research on Innovative Areas (Research in a proposed research area)

Allocation TypeSingle-year Grants
Review Section Biological Sciences
Research InstitutionYokohama City University

Principal Investigator

橋本 博  横浜市立大学, 生命ナノシステム科学研究科, 准教授 (40336590)

Project Period (FY) 2011-04-01 – 2013-03-31
Project Status Completed (Fiscal Year 2012)
Budget Amount *help
¥4,940,000 (Direct Cost: ¥3,800,000、Indirect Cost: ¥1,140,000)
Fiscal Year 2012: ¥2,470,000 (Direct Cost: ¥1,900,000、Indirect Cost: ¥570,000)
Fiscal Year 2011: ¥2,470,000 (Direct Cost: ¥1,900,000、Indirect Cost: ¥570,000)
KeywordsRNA複合体 / 結晶化 / 構造解析 / X線結晶構造解析 / 始原生殖細胞 / RNA / 翻訳 / 相互作用 / 複合体
Outline of Annual Research Achievements

生殖細胞は次世代に遺伝情報を伝え, 種の保存を実現する唯一の細胞であり, 次の世代をつくる全能性を有する. 生殖細胞の前駆細胞である始原生殖細胞の形成と維持にはNanosとPumilioが必要である. NanosとPumilioは, 高等真核生物で保存されたRNA結合タンパク質であり, mRNAの3' 非翻訳領域に結合し, 標的mRNAの翻訳を抑制する. しかし, その制御機構は不明であり, Nanos/Pumilio/mRNA複合体の原子レベルでの立体構造解析が求められている. 本研究は, 生殖細胞の基になる始原生殖細胞の形成と維持に必要なmRNAの翻訳制御因子, NanosとPumilioの構造生物学的研究であり, それらが機能している現場である複合体のX線結晶構造解析を目指す.
前年度にPumilioのC末端ドメイン(PumCTD), NanosのC末端ドメイン(NosCTD)とRNAとの複合体の結晶化に成功したが, 平成24年度は三者複合体のX線結晶構造解析を目指した試料調製を精力的に行った. 10-merと20-merの単鎖RNAを用いて, まずPumCTDとRNAを混合し, その後, NosCTDを添加しゲル濾過を行ったところ, 10-merのRNAでは三者複合体を形成しなかったが, 20-merのRNAを用いた場合に三者複合体としてゲル濾過カラムから溶出することに成功した. また,NosCTDとPumCTDはゲル濾過では相互作用が確認出来なかった. さらに, RNAに対する親和性は, NosCTDよりもPumCTDの方が高いという結果を得ている. これらの結果から, まずPumilioが配列特異的にRNAに結合し, その後NanosがPumilio-RNA複合体に結合し, 三者複合体が形成するモデルを提唱できる.

Research Progress Status

24年度が最終年度であるため、記入しない。

Strategy for Future Research Activity

24年度が最終年度であるため、記入しない。

Report

(2 results)
  • 2012 Annual Research Report
  • 2011 Annual Research Report
  • Research Products

    (34 results)

All 2013 2012 2011

All Journal Article (13 results) (of which Peer Reviewed: 13 results) Presentation (21 results) (of which Invited: 1 results)

  • [Journal Article] Crystal structure of aldoxime dehydratase and its catalytic mechanism involved in carbon-nitrogen triple-bond synthesis2013

    • Author(s)
      Nomura, J., Hashimoto, H., Ohta, T., Hashimoto, Y., Wada, K., Naruta, Y., Oinuma, K-I., & Kobayashi, M.
    • Journal Title

      Proc. Natl. Acad. Sci. USA

      Volume: 110 Issue: 8 Pages: 2810-2815

    • DOI

      10.1073/pnas.1200338110

    • Related Report
      2012 Annual Research Report
    • Peer Reviewed
  • [Journal Article] Mechanistic insights into the activation of Rad51-mediated strand exchange from the structure of a recombination activator, the Swi5-Sfr1 complex2013

    • Author(s)
      N.Kuwabara, Y.Murayama, H.Hashimoto, Y.Kokabu, M.Ikeguchi, M.Sato, K.Mayanagi, Y.Tsutsui, H.Iwasaki,and T.Shimizu
    • Journal Title

      Structure

      Volume: 20 Issue: 3 Pages: 440-449

    • DOI

      10.1016/j.str.2012.01.005

    • Related Report
      2012 Annual Research Report 2011 Annual Research Report
    • Peer Reviewed
  • [Journal Article] Structural basis for the inhibition of xyloglucan-specific endo-β-1,4-glucanase (XEG) by XEG- protein inhibitor2012

    • Author(s)
      Takuya Yoshizawa
    • Journal Title

      J Biol Chem

      Volume: 287 Issue: 22 Pages: 18710-18716

    • DOI

      10.1074/jbc.m112.350520

    • Related Report
      2012 Annual Research Report
    • Peer Reviewed
  • [Journal Article] Crystallization and X-ray diffraction analysis of the ternary complex of the C-terminal domain of human REV1 in complex with REV7 bound to a REV3 fragment involved in translesion DNA synthesis2012

    • Author(s)
      Sotaro Kikuchi
    • Journal Title

      Acta Crystallogr Sect F Struct Biol Cryst Commun

      Volume: 68 Issue: 8 Pages: 962-964

    • DOI

      10.1107/s1744309112032435

    • Related Report
      2012 Annual Research Report
    • Peer Reviewed
  • [Journal Article] Structural basis of recruitment of DNA polymerase z by interaction between REV1 and REV72012

    • Author(s)
      Sotaro Kikuchi
    • Journal Title

      J Biol Chem

      Volume: 287 Issue: 40 Pages: 33847-33852

    • DOI

      10.1074/jbc.m112.396838

    • Related Report
      2012 Annual Research Report
    • Peer Reviewed
  • [Journal Article] Identification of a small molecule PCNA inhibitor that disrupts interactions with PIP-Box proteins and inhibits DNA replication2012

    • Author(s)
      Chandanamali Punchihewa, Akira Inoue, Asami Hishiki, Yoshihiro Fujikawa, Michele Connelly, Benjamin Evison, Youming Shao, Richard Heath, Isao Kuraoka, Patrick Rodrigues, Hiroshi Hashimoto, Masanobu Kawanishi, Mamoru Sato, Takashi Yagi, Naoaki Fujii
    • Journal Title

      J Biol Chem

      Volume: 287 Issue: 17 Pages: 14289-14300

    • DOI

      10.1074/jbc.m112.353201

    • Related Report
      2012 Annual Research Report 2011 Annual Research Report
    • Peer Reviewed
  • [Journal Article] A novel heterotetrameric structure of the crenarchaeal PCNA2-PCNA3 complex2011

    • Author(s)
      A.Kawai, H.Hashimoto, S.Higuchi, M.Tsunoda c, M.Sato, K.T.Nakamura, S.Miyamoto
    • Journal Title

      J.Struct.Biol.

      Volume: 174 Issue: 1 Pages: 443-450

    • DOI

      10.1111/j.1574-6968.2011.02233.x

    • Related Report
      2011 Annual Research Report
    • Peer Reviewed
  • [Journal Article] A novel heterotetrameric structure of the crenarchaeal PCNA2-PCNA3 complex2011

    • Author(s)
      Akito Kawai, Hiroshi Hashimoto, Shigesada Higuchi, Masaru Tsunoda, Mamoru Sato, Kazuo T Nakamura, Shuichi Miyamoto
    • Journal Title

      J Struct Biol

      Volume: 174 Issue: 3 Pages: 443-450

    • DOI

      10.1016/j.jsb.2011.02.006

    • Related Report
      2011 Annual Research Report
    • Peer Reviewed
  • [Journal Article] Crystallization of Bg7S, Purification, crystallization and X-ray diffraction study of basic 7S globulin from soybean2011

    • Author(s)
      T.Yoshizawa, H.Hashimoto, T.Shimizu, M.Yamabe, N.Shichijo, K.Hanada, H.Hirano, M.Sato
    • Journal Title

      Acta Cryst.

      Volume: F67 Issue: 11 Pages: 87-89

    • DOI

      10.1111/j.1742-4658.2011.08111.x

    • Related Report
      2011 Annual Research Report
    • Peer Reviewed
  • [Journal Article] Purification, crystallization and X-ray diffraction study of extracellular dermal glycoprotein from carrot and the inhibition complex it forms with an endo-beta-glucanase from Aspergillus aculeatus2011

    • Author(s)
      T.Yoshizawa, T.Shimizu, H.Hirano, M.Sato, H.Hashimoto
    • Journal Title

      Acta Cryst.

      Volume: F67 Issue: 7 Pages: 830-832

    • DOI

      10.1107/s1744309111020045

    • Related Report
      2011 Annual Research Report
    • Peer Reviewed
  • [Journal Article] Structural basis for matrix metalloproteinase-2 (MMP-2)-selective inhibitory action of β-amyloid precursor protein-derived inhibitor2011

    • Author(s)
      H.Hashimoto, T.Takeuchi, K.Komatsu, K.Miyazaki, M.Sato, S.Higashi
    • Journal Title

      J.Biol.Chem.

      Volume: 286 Issue: 38 Pages: 33236-33243

    • DOI

      10.1074/jbc.m111.264176

    • Related Report
      2011 Annual Research Report
    • Peer Reviewed
  • [Journal Article] The Development of N-α-(2-Carboxyl)benzoyl-N5-(2-fluoro-1-iminoethyl)-L-The Development of N-α-(2-Carboxyl)benzoyl-N5-(2-fluoro-1-iminoethyl)-L-ornithine Amide (o-F-amidine) and N-α-(2-Carboxyl)benzoyl-N5-(2-chloro-1-iminoethyl)-L-ornithine Amide (o-Cl-amidine) As Second Generation Protein Arginine Deiminase (PAD) Inhibitors2011

    • Author(s)
      C.P.Causey, J.E.Jones, J.L.Slack, D.Kamei, L.E.Jones, Jr., V.Subramanian, B.Knuckley, P.Ebrahimi, A.A.Chumanevich, Y.Luo, H.Hashimoto, M.Sato, L.J.Hofseth P.R.Thompson
    • Journal Title

      J.Med.Chem.

      Volume: 54 Issue: 19 Pages: 6919-6935

    • DOI

      10.1021/jm2008985

    • Related Report
      2011 Annual Research Report
    • Peer Reviewed
  • [Journal Article] Fission yeast Swi5-Sfr1 protein complex, an activator of Rad51 recombinase, forms an extremely elongated dogleg-shaped structure2011

    • Author(s)
      Kokabu Y, Murayama Y, Kuwabara N, Oroguchi T, Hashimoto H, Tsutsui Y, Nozaki N, Akashi S, Unzai S, Shimizu T, Iwasaki H, Sato M, Ikeguchi M
    • Journal Title

      J Biol Chem

      Volume: 286 Issue: 50 Pages: 43569-43576

    • DOI

      10.1074/jbc.m111.303339

    • Related Report
      2011 Annual Research Report
    • Peer Reviewed
  • [Presentation] REV1-REV7-REV3三者複合体の結晶構造から明らかになったDNAポリメラーゼζのリクルート機構2013

    • Author(s)
      菊池 壮太郎
    • Organizer
      第30回PFシンポジウム
    • Place of Presentation
      つくば国際会議場, 茨城
    • Year and Date
      2013-03-14
    • Related Report
      2012 Annual Research Report
  • [Presentation] REV1-REV7-REV3三者複合体の結晶構造から明らかになったDNAポリメラーゼζのリクルート機構の構造基盤2012

    • Author(s)
      菊池 壮太郎
    • Organizer
      第35回 日本分子生物学会年会
    • Place of Presentation
      福岡国際会議場, 福岡
    • Year and Date
      2012-12-11
    • Related Report
      2012 Annual Research Report
  • [Presentation] Structural basis of recruitment of DNA polymerase zeta by interaction between REV1 and REV72012

    • Author(s)
      Sotaro Kikuchi
    • Organizer
      AsCA 12/CRYSTAL 28
    • Place of Presentation
      Adelaide Convention Centre, Australia
    • Year and Date
      2012-12-02
    • Related Report
      2012 Annual Research Report
  • [Presentation] Crystal structures of human PCNA in complex with small molecule inhibitors for cancer therapeutics2012

    • Author(s)
      Hiroshi Hashimoto
    • Organizer
      AsCA 12/CRYSTAL 28
    • Place of Presentation
      Adelaide Convention Centre, Australia
    • Year and Date
      2012-12-02
    • Related Report
      2012 Annual Research Report
  • [Presentation] Structural basis of recruitment of DNA polymerase ζ by interaction between REV1 and REV72012

    • Author(s)
      Sotaro Kikuchi
    • Organizer
      The 8th 3R Symposium
    • Place of Presentation
      Awaji Yumebutai International Conference Center, Hyogo
    • Year and Date
      2012-11-25
    • Related Report
      2012 Annual Research Report
  • [Presentation] 損傷乗り越えDNA合成に関わるREV1-REV7-REV3三者複合体の結晶構造解析2012

    • Author(s)
      菊池 壮太郎
    • Organizer
      日本結晶学会年会
    • Place of Presentation
      東北大学 片平キャンパス, 宮城
    • Year and Date
      2012-10-25
    • Related Report
      2012 Annual Research Report
  • [Presentation] REV1-REV7-REV3相互作用の構造基盤2012

    • Author(s)
      橋本 博
    • Organizer
      遺伝研研究集会
    • Place of Presentation
      国立遺伝学研究所, 静岡
    • Year and Date
      2012-10-03
    • Related Report
      2012 Annual Research Report
    • Invited
  • [Presentation] 抗がん剤併用薬開発に向けた PCNAと阻害剤との複合体構造2012

    • Author(s)
      菊池 壮太郎
    • Organizer
      第12回日本蛋白質科学会
    • Place of Presentation
      名古屋国際会議場, 愛知
    • Year and Date
      2012-06-20
    • Related Report
      2012 Annual Research Report
  • [Presentation] ニンジンEDGPの麹菌キシログルカン特異的エンドグルカナーゼ阻害機構2012

    • Author(s)
      吉澤拓也, 清水敏之, 平野久, 橋本博, 佐藤衛
    • Organizer
      第29回PFシンポジウム
    • Place of Presentation
      つくば国際会議場
    • Related Report
      2011 Annual Research Report
  • [Presentation] Structure of REV7 in complex with a REV3 fragment and structural implication of the interaction between DNA polymerase ζ and REV12011

    • Author(s)
      Hiroshi Hashimoto
    • Organizer
      SNY-YCU Bilateral Symposium on Structural Biology
    • Place of Presentation
      RIEKN Yokohama Institute, Yokohama, Japan
    • Year and Date
      2011-11-14
    • Related Report
      2011 Annual Research Report
  • [Presentation] 始原生殖細胞の維持に関わるNanosの構造生物学的研究2011

    • Author(s)
      橋本博, 原幸大, 菱木麻美, 川口茂太, 吉澤拓也, 雲財悟, 田丸浩, 清水敏之, 佐藤衛
    • Organizer
      第11回日本蛋白質科学会
    • Place of Presentation
      ホテル阪急エキスポパーク
    • Related Report
      2011 Annual Research Report
  • [Presentation] 始原生殖細胞の形成に関わるNanosの構造生物学的研究2011

    • Author(s)
      橋本博, 原幸大, 菱木麻美, 川口茂太, 七條直記, 中村敬至, 雲財悟, 田丸浩, 清水敏之, 佐藤衛
    • Organizer
      第28回PFシンポジウム
    • Place of Presentation
      つくば国際会議場
    • Related Report
      2011 Annual Research Report
  • [Presentation] 腸炎ビブリオ由来耐熱性溶血毒TDHの構造2011

    • Author(s)
      橋本博, 中平久美子, 山根努, 福井貴史, 大西紀陽久, 清水敏之, 本田武司, 佐藤衛, 池口満徳, 柳原格
    • Organizer
      第28回PFシンポジウム
    • Place of Presentation
      つくば国際会議場
    • Related Report
      2011 Annual Research Report
  • [Presentation] 制限酵素EcoO109IのDNA結合に関する構造研究2011

    • Author(s)
      渡部みなみ, 橋本博, 喜多恵子, 清水敏之, 佐藤衛
    • Organizer
      第28回PFシンポジウム
    • Place of Presentation
      つくば国際会議場
    • Related Report
      2011 Annual Research Report
  • [Presentation] MMP-2とAPP由来インヒビターペプチドとの複合体の結晶構造解析2011

    • Author(s)
      竹内友香, 橋本博, 宮崎香, 佐藤衛, 東昌市
    • Organizer
      第16回日本病態プロテアーゼ学会学術集会
    • Place of Presentation
      千里ライフサイエンスセンター
    • Related Report
      2011 Annual Research Report
  • [Presentation] Crystal structure of zinc-finger domain of Nanos and its functional implications2011

    • Author(s)
      Hiroshi Hashimoto, Kodai Hara, Asami Hishiki, Shigeta Kawaguchi, Takuya Yoshizawa, Satoru Unzai, Yutaka Tamaru, Toshiyuki Shimizu, Mamoru Sato
    • Organizer
      XXII Congress IUCr
    • Place of Presentation
      Municipal Conference Center, Madrid, Spain
    • Related Report
      2011 Annual Research Report
  • [Presentation] Structural basis of inhibition mechanism by carrot EDGP against GH12 The inhibitory structure of carrot EDGP against endoglucanase from A.aculeatus2011

    • Author(s)
      Takuya Yoshizawa, Toshiyuki Shimizu, Hisashi Hirano, Mamoru Sato, Hiroshi Hashimoto
    • Organizer
      XXII Congress IUCr
    • Place of Presentation
      Municipal Conference Center, Madrid, Spain
    • Related Report
      2011 Annual Research Report
  • [Presentation] Analysis on dynamical structure of intrinsically disordered protein Hef, using MD-SAXS method2011

    • Author(s)
      Tomotaka Oroguchi, Takashi Oda, Hiroshi Hashimoto, Yoshizumi Ishino, Mitsunori Ikeguchi, Mamoru Sato
    • Organizer
      第49回日本生物物理学会年会
    • Place of Presentation
      兵庫県立大学姫路書写キャンパス
    • Related Report
      2011 Annual Research Report
  • [Presentation] APP由来インヒビターペプチドによるMMP-2選択的阻害機構の解明2011

    • Author(s)
      竹内友香, 橋本博, 宮崎香, 佐藤衛, 東昌市
    • Organizer
      第84回日本生化学
    • Place of Presentation
      京都国際会館
    • Related Report
      2011 Annual Research Report
  • [Presentation] Structure of REV7 in complex with a REV3 fragment and structural implication of the interaction between DNA polymerase ζ and REV12011

    • Author(s)
      Hiroshi Hashimoto, Kodai Hara, Yoshiki Murakumo, Shunsuke Kobayashi, Toshiaki Kogame, Satoru Unzai, Satoko Akashi, Shunichi Takeda, Toshiyuki Shimizu, Mamoru Sato
    • Organizer
      Current Opinion in Structural Biology & DNA Repair
    • Place of Presentation
      NH Grand Hotel Krasnapolsky, Amsterdam, Netherlands
    • Related Report
      2011 Annual Research Report
  • [Presentation] ニンジンEDGPによる麹菌XEGの阻害複合体構造2011

    • Author(s)
      吉澤拓也, 橋本博, 清水敏之, 平野久, 佐藤衛
    • Organizer
      平成23年度日本結晶学会年会
    • Place of Presentation
      北海道大学学術交流会館
    • Related Report
      2011 Annual Research Report

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Published: 2011-04-06   Modified: 2018-03-28  

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