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Direct observation of dynein-microtubule binding

Research Project

Project/Area Number 15370064
Research Category

Grant-in-Aid for Scientific Research (B)

Allocation TypeSingle-year Grants
Section一般
Research Field Biophysics
Research InstitutionThe University of Tokyo

Principal Investigator

KAMIYA Ritsu  The University of Tokyo, Department of Biological Sciences, Professor, 大学院・理学系研究科, 教授 (10124314)

Co-Investigator(Kenkyū-buntansha) HIRONO Masafumi  The University of Tokyo, Graduate School of Science, Associate Professor, 大学院・理学系研究科, 助教授 (10212177)
YAGI Tohiki  The University of Tokyo, Graduate School of Science, Research Associate, 大学院・理学系研究科, 助手 (40292833)
Project Period (FY) 2003 – 2004
Project Status Completed (Fiscal Year 2004)
Budget Amount *help
¥9,100,000 (Direct Cost: ¥9,100,000)
Fiscal Year 2004: ¥2,500,000 (Direct Cost: ¥2,500,000)
Fiscal Year 2003: ¥6,600,000 (Direct Cost: ¥6,600,000)
KeywordsCilia and flagella / Dynein arm / Microtubule / Total reflection microscope / Fluorescence label / Tektin
Research Abstract

In thee outer doublet microtubule in the flagellar axoneme, the outer dynein arms are arranged with a 24 nm interval, while each of the seven kinds of inner arms are arranged with a 96 nm interval. How such a regular arrangement is produced is almost entirely unknown. In this study, we aimed to approach this mechanism by directly observing the dynein-microtubule binding process. Using total reflection optics and fluorescently labeled proteins, we observed that the association of dynein to the microtubule is a cooperative process, quantified the cooperative binding using outer-dynein-arm-docking complex (ODA-DC) expressed in insect culture cells. Along with these studies, we searched for proteins responsible for the binding of inner dynein arms and discovered that the protein tektin is involved in the attachment of dynein e in Chlamydomonas axonemes. This is the first study to show tektin involvement in the dynein arrangement ; tektin, first discovered in sea urchin sperm flagella, has been thought to function as a ruler that determines the longitudinal periodicity in the axoneme. In a still different line of experiment, we examined the dynamic nature of dynein-microtubule association using a technique called fluorescence recovery after photobleaching. Using electroporation-mediated incorporation of fluorescently labeled dynein subunit, we found that inner arm dynein of Chlamydomonas is constantly being turned over within the flagella. These findings should provide important insights into the mechanism as to how dynein arms are attached to axonemal microtubules.

Report

(3 results)
  • 2004 Annual Research Report   Final Research Report Summary
  • 2003 Annual Research Report
  • Research Products

    (29 results)

All 2005 2004 2003 Other

All Journal Article (23 results) Publications (6 results)

  • [Journal Article] The mouse ortholog of EFHC1 implicated in juvenile myoclonic epilepsy is an axonemal protein widely conserved among organisms with motile cilia and flagella2005

    • Author(s)
      Ikeda, T.
    • Journal Title

      FEBS Letters 579

      Pages: 819-822

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2004 Final Research Report Summary
  • [Journal Article] Phototactic activity in Chlamydomonas "non-phototactic" mutants deficient in Ca^<2+>-dependent control of flagellar dominance or in inner arm dynein2005

    • Author(s)
      Okita, N.
    • Journal Title

      Journal of Cell Science 118

      Pages: 529-537

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2004 Final Research Report Summary
  • [Journal Article] The mouse ortholog of EFHC1 implicated in juvenile myoclonic epilepsy is an axonemal protein widely conserved among organisms with mottle cilia and flagella2005

    • Author(s)
      Ikeda, T.
    • Journal Title

      FEBS Letters 579

      Pages: 819-822

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2004 Final Research Report Summary
  • [Journal Article] Phototactic activity in Chlamydoinonas "non-phototactic" mutants deficient in Ca^<2+>-dependent control of flagellar dominance or in inner arm dynein2005

    • Author(s)
      Okita, N.
    • Journal Title

      Journal of Cell Science 118

      Pages: 529-537

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2004 Final Research Report Summary
  • [Journal Article] The mouse ortholog of EFHC1 implicated in juvenile myoclonic epilepsy is an axonemal protein widely conserved among organisms with motile cilia andflagella.2005

    • Author(s)
      Ikeda, T.
    • Journal Title

      FEBS Lett. 579

      Pages: 819-822

    • Related Report
      2004 Annual Research Report
  • [Journal Article] Phototactic activity in Chlamydomonas "non-phototactic" mutants deficient in Ca^<2+>-dependent control of flagellar dominance or in inner arm dynein.2005

    • Author(s)
      Okita, N.
    • Journal Title

      J.Cell Sci. 118

      Pages: 529-537

    • Related Report
      2004 Annual Research Report
  • [Journal Article] A tektin homologue is decreased in Chlamydomonas mutants lacking an axonemal inner-arm dynein2004

    • Author(s)
      Yanagisawa, H.
    • Journal Title

      Molecular Biology of the Cell 15

      Pages: 2105-2115

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2004 Final Research Report Summary
  • [Journal Article] Slow ADP-dependent acceleration of microtubule translocation produced by an axonemal dynein2004

    • Author(s)
      Kikushima, K.
    • Journal Title

      FEBS Letters 563

      Pages: 119-122

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2004 Final Research Report Summary
  • [Journal Article] Oda5p, a novel axonemal protein required for assembly of the outer dynein arm and an associated adenylate kinase2004

    • Author(s)
      Blomberg-Wirschell, M.
    • Journal Title

      Molecular Biology of the Cell 15

      Pages: 2729-2741

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2004 Final Research Report Summary
  • [Journal Article] Turnover of actin in Chlamydomonas flagella detected by fluorescence recovery after photobleaching (FRAP)2004

    • Author(s)
      Watanabe, Y.
    • Journal Title

      Cell Structure and Function 29

      Pages: 67-72

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2004 Final Research Report Summary
  • [Journal Article] A tektin homologue is decreased in Chlamzydomonas mutants lacking an axonemal inner-arm dynein2004

    • Author(s)
      Yanagisawa, H.
    • Journal Title

      Molecular Biology of the Cell 15

      Pages: 2105-2115

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2004 Final Research Report Summary
  • [Journal Article] Turnover of actin in Chlamydonzonas flagella detected by fluorescence recovery after photobleaching (FRAP)2004

    • Author(s)
      Watanabe, Y.
    • Journal Title

      Cell Structure and Function 29

      Pages: 67-72

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2004 Final Research Report Summary
  • [Journal Article] A tektin homologue is decreased in Chlamydomonas mutants lacking an axonemal inner-arm dynein.2004

    • Author(s)
      Yanagisawa, H.
    • Journal Title

      Mol.Biol.Cell 15

      Pages: 2105-2115

    • Related Report
      2004 Annual Research Report
  • [Journal Article] Slow ADP-dependent acceleration of microtubule translocation produced by an axonemal dynein.2004

    • Author(s)
      Kikushima, K.
    • Journal Title

      FEBS Lett. 563

      Pages: 119-122

    • Related Report
      2004 Annual Research Report
  • [Journal Article] Oda5p, a novel axonemal protein required for assembly of the outer dynein arm and an associated adenylate kinase.2004

    • Author(s)
      Blomberg-Wirschell, M.
    • Journal Title

      Mol.Biol.Cell 15

      Pages: 2729-2741

    • Related Report
      2004 Annual Research Report
  • [Journal Article] Turnover of actin in Chlamydomonas flagella detected by fluorescence recovery after photobleaching (FRAP).2004

    • Author(s)
      Watanabe, Y.
    • Journal Title

      Cell Struct.Funct. 29

      Pages: 67-72

    • Related Report
      2004 Annual Research Report
  • [Journal Article] Identification of a novel leucine-rich repeat protein as a component of flagellar radial spoke in the ascidian Ciona intestinali2003

    • Author(s)
      Padma, P.
    • Journal Title

      Molecular Biology of the Cell 14

      Pages: 774-785

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2004 Final Research Report Summary
  • [Journal Article] Rib72, a conserved protein associated with the ribbon compartment of flagellar A-microtubules and potentially involved in the linkage between outer doublet microtubules2003

    • Author(s)
      Ikeda, K.
    • Journal Title

      Journal of Biological Chemistry 278

      Pages: 7725-7734

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2004 Final Research Report Summary
  • [Journal Article] DC3, the 21-kD subunit of the outer dynein arm-docking complex (ODA-DC), is a novel EE-hand protein important for assembly of both the outer arm and the ODA-DC2003

    • Author(s)
      Casey, D.M.
    • Journal Title

      Molecular Biology of the Cell 14

      Pages: 3650-3663

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2004 Final Research Report Summary
  • [Journal Article] A novel family of unconventional actin in volvocalean algae2003

    • Author(s)
      Kato-Minoura, T.
    • Journal Title

      Journal of Molecular Evolution 57

      Pages: 555-561

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2004 Final Research Report Summary
  • [Journal Article] DC3, the smallest subunit of the Chlamydomonas flagellar outer dynein arm-docking complex, is a redox-sensitive calcium-binding protein2003

    • Author(s)
      Casey, D.M.
    • Journal Title

      Journal of Biological Chemistry 278

      Pages: 42652-42659

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2004 Final Research Report Summary
  • [Journal Article] DC3, the 21-kD subunit of the outer dynein arm-docking complex (ODA-DC), is a novel EF-hand protein important for assembly of both the outer arm and the ODA-DC2003

    • Author(s)
      Casey, D.M.
    • Journal Title

      Molecular Biology of the Cell 14

      Pages: 3650-3663

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2004 Final Research Report Summary
  • [Journal Article] DC3, the smallest subunit of the Chlamydonzonas flagellar outer dynein arm-docking complex, is a redox-sensitive calcium-binding protein2003

    • Author(s)
      Casey, D.M.
    • Journal Title

      Journal of Biological Chemistry 278

      Pages: 42652-42659

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2004 Final Research Report Summary
  • [Publications] Padma, P.: "Identification of a novel leucine-rich repeat protein as a component of flagellar radial spoke in the ascidian Ciona intesinalis"Molecular Biology of the Cell. 14. 774-785 (2003)

    • Related Report
      2003 Annual Research Report
  • [Publications] Ikeda, K.: "Rib72, a conserved protein associated with the ribbon compartment of flagellar A-microtubules and potentially involved in the linkage between outer doublet microtubules"Journal of Biological Chemistry. 278. 7725-7734 (2003)

    • Related Report
      2003 Annual Research Report
  • [Publications] Casey, D.M.: "DC3, the 21-kD subunit of the outer dynein arm-docking complex (ODA-DC), is a novel EF-hand protein important for assembly of both the outer arm and the ODA-DC"Molecular Biology of the Cell. 14. 3650-3663 (2003)

    • Related Report
      2003 Annual Research Report
  • [Publications] kato-Minoura, T.: "A novel family of unconventional actin in volvocalean algae"Journal of Molecular Evolution. 57. 555-561 (2003)

    • Related Report
      2003 Annual Research Report
  • [Publications] Casey, D.M.: "DC3, the smallest subunit of the Chlamydomonas flagellar outer dynein arm-docking complex, is a redox-sensitive calcium-binding protein"Journal of Biological Chemistry. 278. 42652-42659 (2003)

    • Related Report
      2003 Annual Research Report
  • [Publications] Yanagisawa, H.: "A Tektin Homologue Is Decreased in Chlamydomonas Mutants Lacking an Axonemal Inner-Arm Dynein"Molecular Biology of the Cell. (in press). (2004)

    • Related Report
      2003 Annual Research Report

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Published: 2003-04-01   Modified: 2016-04-21  

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