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Analysis of dynein function by use of mutants and in vitro motility assay systems

Research Project

Project/Area Number 07408034
Research Category

Grant-in-Aid for Scientific Research (A)

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

Principal Investigator

KAMIYA Ritsu  University of Tokyo, Graduate School of Science, Professor, 大学院・理学系研究科, 教授 (10124314)

Co-Investigator(Kenkyū-buntansha) YOSHIMURA Kenjiro  University of Tokyo, Graduate School of Science, Research Associate, 大学院・理学系研究科, 助手 (10230806)
HIRONO Masafumi  University of Tokyo, Graduate School of Science, Lecturer, 大学院・理学系研究科, 講師 (10212177)
真行寺 千佳子  東京大学, 大学院・理学系研究科, 助教授 (80125997)
Project Period (FY) 1995 – 1997
Project Status Completed (Fiscal Year 1997)
Budget Amount *help
¥5,200,000 (Direct Cost: ¥5,200,000)
Fiscal Year 1997: ¥2,300,000 (Direct Cost: ¥2,300,000)
Fiscal Year 1996: ¥2,900,000 (Direct Cost: ¥2,900,000)
Keywordsflagella / dynein / actin / genes / mutation / Chlamydomonas / cell motility / force production / 突然変異体
Research Abstract

The basis for the beating of eukaryotic cilia and flagella is the sliding movement of microtubules driven by dynein, but how undulating movements are produced is not understood. To approach this problem, we carried out two lines of studies aimed at elucidation of the interaction between microtubules and various kinds of dynein. First, we tried to isolate and characterize Chlamydomonas mutants that lack specific dynein subspecies, and assess the property of the missing dyneins by analyzing the motility of the mutants. Second, we isolated specific dynein subspecies, and assayd their properties using in vitro motility systems. Major findings from these studies are as follows : (1) ida5, a mutant missing a subset of inner arm dyneins, was found to have a mutation in the structural gene of conventional actin, such that no conventional actin is produced in the cell. The discovery of this mutant will help to clarify the physiological role of actin in inner-arm dyneins. (2) The force generating properties of dyneins were found to differ greatly between inner and outer arm dyneins ; in particular, the inner arm is more important than the outer arm when axoneme is moving slowly, such as in the process of bend initiation. (3) The activity of dynein is regulated by nucleotides, such as ADP.(4) Outer arm dynein can be self-activated to produce force under various conditions, while inner arm dyneins cannot-they need the central pair and radial spokes for force generation. In short, this study has revealed a great functional diversity among different dyneins. This diversity appears to be important for the axonemal function.

Report

(4 results)
  • 1997 Annual Research Report   Final Research Report Summary
  • 1996 Annual Research Report
  • 1995 Annual Research Report
  • Research Products

    (45 results)

All Other

All Publications (45 results)

  • [Publications] Minoura, I.: "Strikingly different propulsive forces generated by different dynein deficient Chlamydomonas mutants in viscous media" Cell Motil.Cytoskeleton. 31. 130-139 (1995)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] Yagi, T.: "Novel mode of hyper-oscillation in the paralyzed axoneme of a Chlamydomonas mutant lacking the central-pair microtubules" Cell Motil.Cytoskeleton. 31. 207-214 (1995)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] Ogawa, K.: "Interspecies conservation of outer arm dynein intermediate chain sequences defines two intermediate chain subclasses" Molec.Biol.Cell. 6. 685-696 (1995)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] Kamiya, R.: "Exploring the function of inner and outer dynein arms with Chlamydomonasmutants" Cell Motil.Cytoskeleton. 32. 98-102 (1995)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] Kagami, O.: "Nonradioactive method for ATPaseassays" Methods Cell Biol.47. 147-150 (1995)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] Kagami, O.: "Separation of dynein species by high-pressure liquid chromatography" Methods Cell Biol.47. 487-489 (1995)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] Kamiya, R.: "Strategies for isolation of flagellar motility and assembly mutants in Chlamydomonas" Methods Cell Biol.47. 541-544 (1995)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] Nakamura, S.: "Assembly and function of Chlamydomonas flagellar mastigonemes as probed with a monoclonal antibody" J.Cell Sci.109. 57-62 (1996)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] Omoto, C.K.: "Ability of paralyzed flagella mutants of Chlamydomonas to move" Cell Motil.Cytoskeleton. 33. 88-94 (1996)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] Sugase, Y.: "Cloning and characterization of the actin-encoding gene og Chlamydomonasreinhardtii" Gene. 168. 117-121 (1996)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] Takada, S.: "Beat frequency difference between the two flagella of Chlamydomonas depends on the attachment site of outer dynein arms on the outer-doublet microtubules" Cell Motil.Cytoskeleton. 36. 68-75 (1997)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] Kato-Minoura, T.: "Chlamydomonas inner-arm dynein mutant,ida5,has a mutation in an actin-encoding gene" J.Cell Biol.137. 649-656 (1997)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] Hayashibe, K.: "Induction of temporary beating in paralyzed flagella of Chlamydomonas mutants by application of external force" Cell Motil.Cytoskeleton. 37. 232-239 (1997)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] Wakabayashi, K.: "Ca2^+-dependent waveform conversion in the flagellar axoneme of Chlamydomonas mutants lacking the central-pair/radial spoke system" Cell Motil.Cytoskeleton. 38. 22-28 (1997)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] Minoura, I.and Kamiya, R.: "Strikingly different propulsive forces generated by different dynein deficient Chlamydomonas mutants in viscous media" Cell Motil.Cytoskeleton. 31. 130-139 (1995)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] Yagi, T.and Kamiya, R.: "A novel mode of hyper-oscillation in the paralyzed axoneme of a Chlamydomonas mutant lacking the central-pair microtubules." Cell Motil.Cytoskeleton. 31. 207-214 (1995)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] Ogawa, K., Kamiya, R., Wilkerson, C.G., and Witman, G.B.: "Interspecies conservation of outer arm dynein intermediate chain sequences defines two intermediate chain subclasses" Molec.Biol.Cell. 6. 685-696 (1995)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] Kamiya, R.: "Exploring the function of inner and outer dynein arms with Chalamydomonas mutants." Cell Motil.Cytoskeleton. 32. 98-102 (1995)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] Kagami, O.and Kamiya, R.: "Nonradioactive method for ATPase assays." Methods Cell Biol.47. 147-150 (1995)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] Kagami, O.and Kamiya, R.: "Separation of dynein species by high-pressure liquid chromatography." Methods Cell Biol.47. 487-489 (1995)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] Kamiya, R.: "Strategies for isolation of flagellar motility and assembly mutants in Chlamydomonas" Methods Cell Biol.47. 541-544 (1995)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] Nakamura, S., Tanaka, G., Maeda, T., Kamiya, R., Matsunaga, T.and Nikaido, O.: "Assembly and function of Chlamydomonas flagellar mastigonemes as probed with a monoclonal antibody." J.Cell Sci.109. 57-62 (1996)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] Sugase, Y., Hirono, M., Kindle, K.L., and Kamiya, R.: "Cloning and characterization of the actin-encoding gene of Chlamydomonas reinhardtii." Gene 168. 117-121 (1996)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] Omoto, C.K., Yagi, T., Kurimoto, E., and Kamiya, R.: "Ability of paralyzed flagella mutants of Chlamydomonas to move" Cell Motil.Cytoskeleton. 33. 88-94 (1996)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] Takada, S.and Kamiya, R.: "Beat frequency difference between the two flagella of Chlamydomonas depends on the attachment site of outer dynein arms on the outer doublet microtubules." Cell Motil.Cytoskeleton. 36. 68-75 (1997)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] Kato-Minoura, T., Hirono, M., and Kamiya, R.: "Chlamydomonas inner-arm dynein mutant, ide5, has a mutation in an actin encoding gene." J.Cell Biol.137. 68-75 (1997)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] Hayashibe, K., Shingyoji, C.and Kamiya, R.: "Induction of temporary beating in paralyzed flagella of Chlamydomonas mutants by application of external force" Cell Motil.Cytoskeleton. 37. 232-239 (1997)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] Wakabayashi, K., Yagi, T.and Kamiya, R.: "Ca^<2+>-dependent waveform conversion in the flagellar axoneme of Chlamydomonas mutants lacking the central-pair/radial spoke system" Cell Motil.Cytoskeleton. 38. 22-28 (1997)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] Takada,S.: "Beat frequency difference between the two flagella of Chlamydomonas depends on the attachment site of outer dynein arms on the outer-doublet microtubules" Cell Motil.Cytoskeleton. 36. 68-75 (1997)

    • Related Report
      1997 Annual Research Report
  • [Publications] Kato-Minoura,T.: "Chlamydomonas inner-arm dynein mutant,ida5,has a mutation in an actin encoding gene." J.Cell Biol.137. 649-656 (1997)

    • Related Report
      1997 Annual Research Report
  • [Publications] Hayashibe,K.: "Induction of temporary beating in paralyzed flagella of Chlamydomonas mutants by application of external force" Cell Motil.Cytoskeleton. 37. 232-239 (1997)

    • Related Report
      1997 Annual Research Report
  • [Publications] Wakabayashi,K.: "Ca2+-dependent waveform conversion in the flagellar axoneme of Chlamydomonas mutants lacking the central-pair/radial spoke system" Cell Motil.Cytoskeleton. 38. 22-28 (1997)

    • Related Report
      1997 Annual Research Report
  • [Publications] Omoto,C.K.: "Ability of paralyzed flagella mutants of Chlamydomonas to move" Cell Motility and the Cytoskeleton. 33. 88-94 (1996)

    • Related Report
      1996 Annual Research Report
  • [Publications] Sugase,Y.: "Cloning and characterization of the actin-encoding gene of Chlamydomonas reinhardtii" Gene. 168. 117-121 (1996)

    • Related Report
      1996 Annual Research Report
  • [Publications] Yoshimura,K.: "A Novel Type of Mechanoreception by the Flagella of Chlamydomonas" Journal of Exprimental Biology. 199. 295-302 (1996)

    • Related Report
      1996 Annual Research Report
  • [Publications] Takada,S.: "Beat frequency difference between the two flagella of Chlamydomonas depends on the attachment site of outer dynein arms on the outer-doublet microtubules" Cell Motilility and the Cytoskeleton. 36. 68-75 (1997)

    • Related Report
      1996 Annual Research Report
  • [Publications] Yoshimura,K.: "Conversion of beating mode in Chlamydomonas flagella induced by electric stimulation" Cell Motility and the Cytoskeleton. 36(印刷中). (1997)

    • Related Report
      1996 Annual Research Report
  • [Publications] Minoura,I.: "Strikingly different propulsive forces generated by different dynein deficient Chlamydomonas mutants in viscous media" Cell Motil.Cytoskeleton. 31. 130-139 (1995)

    • Related Report
      1995 Annual Research Report
  • [Publications] Yagi,T.: "Novel mode of hyper-oscillation in the paralyzed axoneme of a Chlamydomonas mutant lacking the centralpair microtubules" Cell Motil.Cytoskeleton. 31. 207-214 (1995)

    • Related Report
      1995 Annual Research Report
  • [Publications] Ogawa,K.: "Interspecies conservation of outer arm dynein intermediae chain sequences defines two intermediate chain subclasses" Molec.Biol.Cell. 6. 685-696 (1995)

    • Related Report
      1995 Annual Research Report
  • [Publications] Kamiya,R.: "Exploring the function of inner and outer dynein arms with Chlamydomonas mutants" Cell Motil.Cytoskeleton. 32. 98-102 (1995)

    • Related Report
      1995 Annual Research Report
  • [Publications] Nakamura,S.: "Assembly and function of Chlamydomonas flagellar mastigonemes as probed with a monoclonal antibody" J.Cell Sci.109. 57-62 (1996)

    • Related Report
      1995 Annual Research Report
  • [Publications] Omoto,C.K.: "Ability of paralyzed flagella mutants of Chlamydomonas to move" Cell Motil.Cytoskeleton. (印刷中).

    • Related Report
      1995 Annual Research Report
  • [Publications] Sugase,Y.: "Cloning and characterization of the actin-encoding gene of Chlamydomonas reinhardtii" Gene. (印刷中).

    • Related Report
      1995 Annual Research Report
  • [Publications] Yoshimura,K.: "A Novel Type of Mechanoreception by the Flagella of Chlamydomonas." J.Exp.Biol.(印刷中).

    • Related Report
      1995 Annual Research Report

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Published: 1996-04-01   Modified: 2016-04-21  

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