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Roles of Chk1 phosphorylation in DNA damage checkpoint

Research Project

Project/Area Number 20790096
Research Category

Grant-in-Aid for Young Scientists (B)

Allocation TypeSingle-year Grants
Research Field Biological pharmacy
Research InstitutionAichi Cancer Center Research Institute

Principal Investigator

KOUSUKE Kasahara  Aichi Cancer Center Research Institute, 発がん制御研究部, 研究員 (90455535)

Project Period (FY) 2008 – 2009
Project Status Completed (Fiscal Year 2009)
Budget Amount *help
¥4,290,000 (Direct Cost: ¥3,300,000、Indirect Cost: ¥990,000)
Fiscal Year 2009: ¥2,080,000 (Direct Cost: ¥1,600,000、Indirect Cost: ¥480,000)
Fiscal Year 2008: ¥2,210,000 (Direct Cost: ¥1,700,000、Indirect Cost: ¥510,000)
Keywordsチェックポイント / DNA損傷 / リン酸化シグナル伝達 / 細胞周期 / 分指標的治療薬 / Chk1 / DNAダメージ / キナーゼ / リン酸化修飾 / リン酸化
Research Abstract

Chk1は、DNA障害チェックポイントの際にATRキナーゼによってセリン317及びセリン345がリン酸化される。このリン酸化によって、Chk1は立体構造が変化し、Chk1のキナーゼドメインと基質間の相互作用が亢進すると考えられているが、Chk1の詳細な活性制御メカニズムは未だ不明な点が多い。今回、我々は機能未知であるChk1のリン酸化反応からChk1の機能制御にアプローチし、以下のことを明らかにした。
1. DNA障害チェックポイントにおいて、ATRによってリン酸化されたChk1は活性化し、その後、自身のセリン296をリン酸化(自己リン酸化)することを明らかにした。また、セリン296がリン酸化されたChk1がドッキング蛋白質14-3-3のγサブタイプと直接結合すること、そして2量体形成した14-3-3がさらにCdc25Aと結合することで、Chk1/14-3-3γ/Cdc25Aの3者複合体が形成されることを見出した。この複合体形成により、Chk1はCdc25Aのセリン76をリン酸化し、Cdc25Aの蛋白質分解を誘導することが判明した。
2. 分裂期においてChk1はセリン286及びセリン301をサイクリン依存性キナーゼ1(CDK1)にリン酸化されることを同定した。さらに、このリン酸化は、分裂前期(prophase)においてChk1を核から細胞質へ排出を促すことを明らかにした。この機構により、細胞の分裂期進行が促進される。また、Chk1のセリン286及びセリン301はDNA障害チェックポイントにおいては、CDK2によってリン酸化されることも見出した。

Report

(3 results)
  • 2009 Annual Research Report   Final Research Report ( PDF )
  • 2008 Annual Research Report
  • Research Products

    (39 results)

All 2010 2009 2008 Other

All Journal Article (18 results) (of which Peer Reviewed: 8 results) Presentation (17 results) Book (4 results)

  • [Journal Article] Withaferin A targets intermediate filaments GFAP and vimentin in A model of retinal gliosis.2010

    • Author(s)
      P. Bargagna-Mohan, R.R. Paranthan, A. Hamza, N. Dimova, B. Trucchi, C. Srinivasan, G.I. Elliott, C.G. Zhan, D.L. Lau, H. Zhu, K. Kasahara, M. Inagaki, F. Cambi, R. Mohan
    • Journal Title

      J Biol Chem. 285

      Pages: 7657-7669

    • Related Report
      2009 Final Research Report
  • [Journal Article] Withaferin a targets intermediate filaments glial fibrillary acidic protein and vimentin in a model of retinal gliosis2010

    • Author(s)
      P.Bargagna-Mohan, et al.
    • Journal Title

      The Journal of Biological Chemistry VOL.285

      Pages: 7657-7669

    • Related Report
      2009 Annual Research Report
    • Peer Reviewed
  • [Journal Article] A novel positive feedback loop between Cycline-dependent kinase 1 (CDK1) ad CHK1 in the nucleus during the G2/M transition.2009

    • Author(s)
      M. Enomoto, H. Goto, Y. Tomono, K. Kasahara, K. Tsujimura, T. Kiyono, M. Inagaki
    • Journal Title

      J. Biol. Chem. 284

      Pages: 34223-3433

    • Related Report
      2009 Final Research Report
  • [Journal Article] Requirement of the SH4 and tyrosine-kinase domains but not the kinase activity of Lyn for its biosynthetic targeting to caveolin-positive Golgi membranes.2009

    • Author(s)
      K. Ikeda, Y. Nakayama, M. Ishii, Y. Obata, K. Kasahara, Y. Fukumoto, N. Yamaguchi
    • Journal Title

      Biochim. Biophys. Aca. 1790

      Pages: 1345-1352

    • Related Report
      2009 Final Research Report
  • [Journal Article] Differential trafficking of c-Src, Lyn, c-Yes, and Fyn is specified by the state of palmitoylation in the SH4 domain.2009

    • Author(s)
      I. Sato, Y. Obata, K. Kasahara, Y. Nakayama, Y. Fukumoto, T. Yamasaki, K.K. Yokoyama, N. Yamaguchi
    • Journal Title

      J. Cell Sci. 122

      Pages: 965-975

    • Related Report
      2009 Final Research Report
  • [Journal Article] Nuclear localization of Src-family tyrosine kinase is required for growth factor-induced euchromatinization.2009

    • Author(s)
      A. Takahashi, Y. Obata, Y. Fukumoto, Y, Nakayama, K. Kasahara, T. Kuga, Y. Higashiyama, T. Saito, K.K. Yokoyama, N. Yamaguchi
    • Journal Title

      Exp. Cell. Re. 315

      Pages: 1117-1141

    • Related Report
      2009 Final Research Report
  • [Journal Article] Novel positive feedback between Cdk1 and Chk1 in the nucleus during G2/M transition2009

    • Author(s)
      Masato Enomoto, et al.
    • Journal Title

      The Journal of Biological Chemistry VOL.284

      Pages: 34223-34030

    • Related Report
      2009 Annual Research Report
    • Peer Reviewed
  • [Journal Article] Differential traffcking of Ser, Lyn, Yes and Fyn is specified by the palmitoylation in the SH4 domain2009

    • Author(s)
      Izumi Sato, et al.
    • Journal Title

      Journal of Cell Science VOL.122

      Pages: 965-975

    • Related Report
      2009 Annual Research Report
    • Peer Reviewed
  • [Journal Article] Requirement of the SH4 and tyrosine-kinase domains but not the kinase activity of Lyn for its biosynthetic targeting to caveolin-positive Golgi membrane2009

    • Author(s)
      Kikuko Ikeda, et al.
    • Journal Title

      Biochimica et Biophysica Acta (BBA) VOL.1790

      Pages: 1345-1352

    • Related Report
      2009 Annual Research Report
    • Peer Reviewed
  • [Journal Article] Nuclear localization of Src-family tyrosine kinase is required for growth factor-induced euchromatinization2009

    • Author(s)
      Akinori Takahashi, et al.
    • Journal Title

      Experimental Cell Research VOL.315

      Pages: 1117-1141

    • Related Report
      2009 Annual Research Report
    • Peer Reviewed
  • [Journal Article] Chk1 phosphorylation at Ser286 and Ser301 occurs with both stalled DNA replication and damage checkpoint stimulation.2008

    • Author(s)
      Y. Ikegami, H. Goto, T. Kiyono, M. Enomoto, K Kasahara, Y. Tomono, K. Tozawa, A. Morita, K. Kohri, M. Inagaki
    • Journal Title

      Biochem. Biophys. Res. Commun. 377

      Pages: 1227-1231

    • Related Report
      2009 Final Research Report
  • [Journal Article] Nuclear localization of Lyn tyrosine kinase mediated by inhibition of its kinase activity.2008

    • Author(s)
      K. Ikeda, Y. Nakayama, Y. Togashi, Y. Obata, T. Kuga, K. Kasahara, Y. Fukumoto, N. Yamaguchi.
    • Journal Title

      Exp. Cell Re. 314

      Pages: 3392-3404

    • Related Report
      2009 Final Research Report
  • [Journal Article] v-Src and c-Src, nonpalmitoylated Src-family kinases, control lysosome distribution via Rab7 in an SH2 domain-dependent manner.2008

    • Author(s)
      K. Kasahara, Y. Nakayama, N. Yamaguchi.
    • Journal Title

      Caner Lett. 262

      Pages: 19-27

    • Related Report
      2009 Final Research Report
  • [Journal Article] Nuclear localization of magphinins, alternative splicing products of the human trophinin gene.2008

    • Author(s)
      J. Aoyama, Y. Akazawa, K. Kasahara, Y. Higashiyama, I. Kikuchi, Y. Fukumoto, S. Saburi, Y. Nakayama, M.N. Fukuda, N. Yamaguchi.
    • Journal Title

      J. Cel. Biochem. 103

      Pages: 765-777

    • Related Report
      2009 Final Research Report
  • [Journal Article] Role of Src-family kinases in formation of the cortical actin cap at the dorsal cell surface.2008

    • Author(s)
      T. Kuga, M. Hoshino, Y. Nakayama, K. Kasahara, K. Ikeda, Y. Obata, A. Takahashi, Y. Higashiyama, Y, Fukumoto, N. Yamaguchi
    • Journal Title

      Exp. Cell Re. 314

      Pages: 2040-2054

    • Related Report
      2009 Final Research Report
  • [Journal Article] v-Src and c-Src, nonpalmitoylated Src-family kinases, control lysosome distribution via Rab7 in SH2 domain-dependent manner2008

    • Author(s)
      Kousuke Kasahara
    • Journal Title

      Cancer Letters 262

      Pages: 19-27

    • Related Report
      2008 Annual Research Report
    • Peer Reviewed
  • [Journal Article] hk1 phosphorylation at Ser286 and Ser301 occurs with both stalled DNA replication and damage checkpoint stimulation2008

    • Author(s)
      Yosuke Ikegami
    • Journal Title

      Biochemical Biophysical Research Communications 377

      Pages: 1227-1231

    • Related Report
      2008 Annual Research Report
    • Peer Reviewed
  • [Journal Article] Nuclear localization of Lyn tyrosine kinase mediated by inhibition of its kinase activity2008

    • Author(s)
      Kikuko Ikeda
    • Journal Title

      Experimental Cell Research 314

      Pages: 3392-3404

    • Related Report
      2008 Annual Research Report
    • Peer Reviewed
  • [Presentation] G2/M期におけるCdk1によるChk1の制御機構の解析2009

    • Author(s)
      榎本将人
    • Organizer
      第81回日本生化学会大会
    • Place of Presentation
      神戸・神戸ポートアイランド
    • Year and Date
      2009-10-27
    • Related Report
      2009 Annual Research Report
  • [Presentation] 多彩なリン酸化によってチェックポイントキナーゼ1(Chk1)は機能修飾される2009

    • Author(s)
      笠原広介
    • Organizer
      第68回日本癌学会学術総会
    • Place of Presentation
      横浜・パシフィコ横浜
    • Year and Date
      2009-10-02
    • Related Report
      2009 Annual Research Report
  • [Presentation] The association between autophosphorylated Chk1 and 14-3-3 gamma is required for Chk1 function in DNA damage checkpoint2009

    • Author(s)
      笠原広介
    • Organizer
      第61回日本細胞生物学会大会
    • Place of Presentation
      名古屋・名古屋国際会議場
    • Year and Date
      2009-06-03
    • Related Report
      2009 Annual Research Report
  • [Presentation] 多彩なリン酸化によってチェックポイントキナーゼ1(Chk1)は機能修飾される2009

    • Author(s)
      笠原広介、後藤英仁、稲垣昌樹
    • Organizer
      第68回日本癌学会学術総会
    • Place of Presentation
      横浜
    • Related Report
      2009 Final Research Report
  • [Presentation] Regulatory mechanism of Chk1 by its autophosphoryaltion in checkpoint response2009

    • Author(s)
      Kasahara, K.
    • Organizer
      Global Center of Excellence (COE) Program 1st International Symposium "Signaling of Cancer Cells"
    • Place of Presentation
      Nagoya
    • Related Report
      2009 Final Research Report
  • [Presentation] 14-3-3γmediates Cdc25A proteolysis to induce S and G2 arrest after DNA damage.2009

    • Author(s)
      Kasahara, K.
    • Organizer
      Global Center of Excellence (COE) Program 2nd International Symposium "Signaling of Cancer Cells"
    • Place of Presentation
      Nagoya
    • Related Report
      2009 Final Research Report
  • [Presentation] 14-3-3γmediates Cdc25A proteolysis to induce S and G2 arrest after DNA damage.2009

    • Author(s)
      Goto, H., Kasahara, K., Tomono, Y., Enomoto, M., Kiyono, T., Inagaki, M.
    • Organizer
      The 49th Annual Meeting of the American Society for Cell Biology
    • Place of Presentation
      San Diego
    • Related Report
      2009 Final Research Report
  • [Presentation] The association between autophosphorylated Chk1 and 14-3-3gamma is required for Chk1 function in DNA damage checkpoint2009

    • Author(s)
      笠原広介, 後藤英仁, 榎本将人, 友野靖子, 清野透, 稲垣昌樹
    • Organizer
      第61回日本細胞生物学会大会
    • Place of Presentation
      名古屋
    • Related Report
      2009 Final Research Report
  • [Presentation] G2/M期におけるCdk1によるChk1の制御機構の解析2009

    • Author(s)
      榎本将人, 後藤英仁, 友野靖子, 笠原広介, 辻村邦夫, 清野透, 稲垣昌樹
    • Organizer
      第82回日本生化学大会
    • Place of Presentation
      神戸
    • Related Report
      2009 Final Research Report
  • [Presentation] Negative feedback regulation of Chk1 by its own kinase acitivity2008

    • Author(s)
      笠原広介、後藤英仁、榎本将人、池上要介、友野靖子、稲垣昌樹
    • Organizer
      2008年BMB2008(第31回日本分子生物学会年会・第81回日本生化学大会合同大会
    • Place of Presentation
      神戸
    • Related Report
      2009 Final Research Report
  • [Presentation] Negative feedback regulation of Chk1 by its own kinase activity2008

    • Author(s)
      笠原広介、後藤英仁、榎本将人、池上要介、友野靖子、稲垣昌樹
    • Organizer
      第31回日本分子生物学会年会・第81回日本生化学大会合同大会
    • Place of Presentation
      神戸
    • Related Report
      2009 Final Research Report 2008 Annual Research Report
  • [Presentation] Chk1 phosphorylaiton by Cyclin-dependent kinase 1 promotes mitotic entry2008

    • Author(s)
      Goto H., Enomoto M., Tomono Y., Kasahara K., Ikegami Y., Tsujimura K., Kiyono T., Inagaki M.
    • Organizer
      2008 American Society for Cell Biology
    • Related Report
      2009 Final Research Report
  • [Presentation] Ser-296 phosphorylation of Chk1 in checkpoint response2008

    • Author(s)
      笠原広介、後藤英仁、池上要介、榎本将人、友野靖子、稲垣昌樹
    • Organizer
      2008年第67回日本癌学会学術総会
    • Place of Presentation
      名古屋
    • Related Report
      2009 Final Research Report
  • [Presentation] Autophosphorylation of Chk1 in checkpoint response2008

    • Author(s)
      笠原広介、後藤英仁、池上要介、榎本将人、友野靖子、稲垣昌樹
    • Organizer
      2008年第60回日本細胞生物学会
    • Place of Presentation
      横浜
    • Related Report
      2009 Final Research Report
  • [Presentation] 細胞増殖因子刺激により誘導されるユークロマチン化における核局在Src型チロシンキナーゼの役割2008

    • Author(s)
      高橋明格、福本泰典、中山祐治、笠原広介、久家貴寿、小幡裕希、東山幸弘、山口直人
    • Organizer
      第31回日本分子生物学会年会・第81回日本生化学大会合同大会
    • Place of Presentation
      神戸
    • Related Report
      2009 Final Research Report
  • [Presentation] Chk1 phosphorylation by Cyclin-dependent kinases (Cdks)

    • Author(s)
      Enomoto, M., Goto, H., Ikegami, Y., Kasahara, K., Tomono, Y., Tsujimura, K., Kiyono, T., Inagaki, M.
    • Organizer
      International Symposium on Chromosome Dynamics in Ise
    • Place of Presentation
      Shima
    • Related Report
      2009 Final Research Report
  • [Presentation] Chk1 phosphorylation by Cyclin-dependent kinase 1 promotes mitotic entry

    • Author(s)
      Goto, H., Enomoto, M. Tomono, Y., Kasahara, K., Ikegami, Y., Tsujimura, K., Kiyono, T. Inagaki, M.
    • Organizer
      The 48th Annual Meeting of the American Society for Cell Biology
    • Place of Presentation
      San Francisco
    • Related Report
      2009 Final Research Report
  • [Book] リン酸化によるM期制御-染色体サイクルゲノム恒常性維持、継承とダイナミクス-正井久雄、升方久夫、釣本敏樹、仁木宏典、篠原彰編2009

    • Author(s)
      笠原広介、後藤英仁
    • Related Report
      2009 Final Research Report
  • [Book] 蛋白質・核酸・酵素染色体サイクル(2009年増刊号;54巻5号)2009

    • Author(s)
      笠原広介
    • Publisher
      共立出版
    • Related Report
      2009 Annual Research Report
  • [Book] 細胞の構造と分裂終期-図説分子病態学-一瀬白帝、鈴木宏治編2008

    • Author(s)
      笠原広介、後藤英仁、稲垣昌樹
    • Publisher
      中外医学社
    • Related Report
      2009 Final Research Report
  • [Book] 図説 分子病態学4版2008

    • Author(s)
      笠原広介
    • Publisher
      中外医学社
    • Related Report
      2008 Annual Research Report

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Published: 2008-04-01   Modified: 2016-04-21  

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