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Roles of cofilin in regulation of actin cytoskeleton dynamics

Research Project

Project/Area Number 17570152
Research Category

Grant-in-Aid for Scientific Research (C)

Allocation TypeSingle-year Grants
Section一般
Research Field Cell biology
Research InstitutionTohoku University

Principal Investigator

OHASHI Kazumasa  Tohoku University, Graduate School of Life Sciences, Associate Professor, 大学院生命科学研究科, 助教授 (10312539)

Project Period (FY) 2005 – 2006
Project Status Completed (Fiscal Year 2006)
Budget Amount *help
¥3,500,000 (Direct Cost: ¥3,500,000)
Fiscal Year 2006: ¥1,500,000 (Direct Cost: ¥1,500,000)
Fiscal Year 2005: ¥2,000,000 (Direct Cost: ¥2,000,000)
KeywordsActin cytoskeleton / cofilin / LIM-kinase / Slingshot / GFP / Phosphorylation / Inhibitor / イメージング / ダイナミクス
Research Abstract

Actin cytoskeletal dynamics and reorganization play essential roles in various intracellular events, including cell migration, cytokinesis, endocytosis, and polarity formation. Actin dynamics and reorganization are spatiotemporally regulated by a large number of actin-binding proteins and upstream signaling molecules. Cofilin, a key regulator of actin filament dynamics, stimulates depolymerization and severing of actin filaments. Cofilin is inactivated by phosphorylation of Ser-3 by LIM-kinase (LIMK). The Ser-3-phosphorylated cofilin (P-cofilin) is dephosphorylated and reactivated by the Slingshot phosphatase. The purposes of this project were to reveal the function of cofilin in actin cytoskeleton reorganization. In the present study, we have developed a new probe that detects the actin-binding activity of cofilin using a bimolecular fluorescence complementation (BiFC) method. The probe was improved higher intensity and sensitivity in this study. We expressed and purified these probe proteins in insect cells using a baculovirus expressing system, and developed methods to measure the actin-binding activity of cofilin, the kinase activity of LIMK, and the phosphatase activity of Slingshot with the fluorescence of complemented-GFP. We screened the inhibitors of cofilin, LIMK and Slingshot with over five hundred chemical compounds. As a consequence, LIMK was inhibited two compounds which was known as Src tyrosine kinase inhibitor. The compounds inhibited the hyper-polymerization of actin in LIMK overexpressing cells. It was reported that one of the compounds inhibited cell migration and neurite extension at the concentration of LIMK inhibition. We are tring to analyze the role of cofilin phosphorylation in the actin cytoskeleton reorganization using the compounds in living cells. We are going to screen more specific inhibitors for LIMK in derivative forms of the compounds. These inhibitors will be seeds of the drugs for inhibition of metastasis and inflammation.

Report

(3 results)
  • 2006 Annual Research Report   Final Research Report Summary
  • 2005 Annual Research Report
  • Research Products

    (23 results)

All 2007 2006 2005

All Journal Article (21 results) Book (2 results)

  • [Journal Article] Actin filaments-stabilizing and -bundling activities of cofilin-phosphatase Slingshot-12007

    • Author(s)
      Kurita, S. et al.
    • Journal Title

      Genes Cells 12・5(印刷中)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2006 Annual Research Report 2006 Final Research Report Summary
  • [Journal Article] Cofilin critically contributes to stimulus-induced lamellipodium formation by generating an abundant supply of actin monomers in the cytoplasm2007

    • Author(s)
      Kiuchi, T. et al.
    • Journal Title

      J. Cell Biol. 177・3

      Pages: 465-476

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2006 Final Research Report Summary
  • [Journal Article] LIM-kinase and slingshot are critical for neurite extension2007

    • Author(s)
      Endo, M. et al.
    • Journal Title

      J. Biol. Chem. 282・18

      Pages: 13692-13702

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2006 Final Research Report Summary
  • [Journal Article] Actin filaments-stabilizing and -bundling activities of cofilin-phosphatase Slingshot-1.2007

    • Author(s)
      Kurita, S. et al.
    • Journal Title

      Genes Cells (in press)

      Pages: 12-5

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2006 Final Research Report Summary
  • [Journal Article] Cofilin critically contributes to stimulus-induced lamellipodium formation by generating an abundant supply of actin monomers in the cytoplasm.2007

    • Author(s)
      Kiuchi, T. et al.
    • Journal Title

      J. Cell Biol. 177-3

      Pages: 465-476

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2006 Final Research Report Summary
  • [Journal Article] LIM-kinase and slingshot are critical for neurite extension.2007

    • Author(s)
      Endo, M. et al.
    • Journal Title

      J. Biol. Chem. 282-18

      Pages: 13692-13702

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2006 Final Research Report Summary
  • [Journal Article] Cofilin critically contributes to stimulus-induced lamellipodium formation by generating an abundant supply of actin monomers in the cytoplasm2007

    • Author(s)
      Kiuchi, T. et al.
    • Journal Title

      J. Cell Biol. 印刷中

    • Related Report
      2006 Annual Research Report
  • [Journal Article] LIM-kinase and slingshot are critical for neurite extension2007

    • Author(s)
      Endo, M. et al.
    • Journal Title

      J. Biol. Chem. 印刷中

    • Related Report
      2006 Annual Research Report
  • [Journal Article] Identification of multiple actin-binding sites in cofilin-phosphatase Slingshot-1L.2006

    • Author(s)
      Yamamoto M. et al.
    • Journal Title

      FEBS Lett. 580・7

      Pages: 1789-1794

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2006 Final Research Report Summary
  • [Journal Article] MAPKAPK-2-mediated LIM-kinase activation is critical for VEGF-induced actin remodeling and cell migration.2006

    • Author(s)
      Kobayashi M. et al.
    • Journal Title

      EMBO J. 25・4

      Pages: 713-726

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2006 Final Research Report Summary
  • [Journal Article] Actin filaments-stabilizing and -bundling activities of cofilin-phosphatase Slingshot-12006

    • Author(s)
      Yamamoto M. et al.
    • Journal Title

      FEBS Lett. 580-7

      Pages: 1789-1794

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2006 Final Research Report Summary
  • [Journal Article] Spatial and temporal regulation of cofilin activity by LIM kinase and Slingshot is critical for directional cell migration.2006

    • Author(s)
      Kobayashi M. et al.
    • Journal Title

      EMBO J. 25-4

      Pages: 713-726

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2006 Final Research Report Summary
  • [Journal Article] Spatial and temporal regulation of cofilin activity by LIM kinase and Slingshot is critical for directional cell migration.2006

    • Author(s)
      Nishita M. et al.
    • Journal Title

      J. Cell Biol. 171-2

      Pages: 349-359

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2006 Final Research Report Summary
  • [Journal Article] The signal transduction of actin cytoskeleton reorganization (ed. by Sugamura, K.)2006

    • Author(s)
      Ohashi, K., et al.
    • Journal Title

      Gene and Medicine MOOK 6

      Pages: 317-317

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2006 Final Research Report Summary
  • [Journal Article] Identification of multiple actin-binding sites in cofilin-phosphatase Slingshot-1L.2006

    • Author(s)
      Yamamoto M.et al.
    • Journal Title

      FEBS Lett. (In press)

    • Related Report
      2005 Annual Research Report
  • [Journal Article] MAPKAPK-2-mediated LIM-kinase activation is critical for VEGF-induced actin remodeling and cell migration.2006

    • Author(s)
      Kobayashi M.et al.
    • Journal Title

      EMBO J. 25・4

      Pages: 713-726

    • Related Report
      2005 Annual Research Report
  • [Journal Article] Spatial and temporal regulation of cofilin activity by LIM kinase and Slingshot is critical for directional cell migration.2005

    • Author(s)
      Nishita M. et al.
    • Journal Title

      J. Cell Biol. 171・2

      Pages: 349-359

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2006 Final Research Report Summary
  • [Journal Article] Alteration of phosphatidylinositol 3-kinase cascade in the multilobulated nuclear formation of adult T cell leukemia/lymphoma (ATLL).2005

    • Author(s)
      Fukuda R. et al.
    • Journal Title

      Proc. Natl. Acad. Sci. USA. 102・42

      Pages: 15213-15218

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2006 Final Research Report Summary
  • [Journal Article] Alteration of phosphatidylinositol 3kinase cascade in the multilobulated nuclear formation of adult T cell leukemia/lymphoma (ATLL).2005

    • Author(s)
      Fukuda R. et al.
    • Journal Title

      Proc. Natl. Acad. Sci. USA. 102-42

      Pages: 15213-15218

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2006 Final Research Report Summary
  • [Journal Article] Spatial and temporal regulation of cofilin activity by LIM kinase and Slingshot is critical for directional cell migration.2005

    • Author(s)
      Nishita M.et al.
    • Journal Title

      J.Cell Biol. 171・2

      Pages: 349-359

    • Related Report
      2005 Annual Research Report
  • [Journal Article] Alteration of phosphatidylinositol 3-kinase cascade in the multilobulated nuclear formation of adult T cell leukemia/lymphoma(ATLL).2005

    • Author(s)
      Fukuda R.et al.
    • Journal Title

      Proc.Natl.Acad.Sci.USA. 102・4

      Pages: 15213-15218

    • Related Report
      2005 Annual Research Report
  • [Book] アクチン骨格の再構築制御とシグナル伝達(遺伝子医学MOOK6シグナル伝達病を知る)(菅村和夫編)2006

    • Author(s)
      大橋一正, 水野健作(分担, 1項目)
    • Total Pages
      317
    • Publisher
      メディカルドゥ
    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2006 Final Research Report Summary
  • [Book] 遺伝子医学MOOK6 シグナル伝達病を知る「アクチン骨格の再構築制御とシグナル伝達」(菅村和夫編)2006

    • Author(s)
      大橋一正, 水野健作(分担、1項目)
    • Total Pages
      317
    • Publisher
      メディカルドゥ
    • Related Report
      2006 Annual Research Report

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Published: 2005-04-01   Modified: 2016-04-21  

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