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Molecular Mechanism of energy transduction of dynein revealed by genetic engineering, structural and enzyme kinetics studies

Research Project

Project/Area Number 17107003
Research Category

Grant-in-Aid for Scientific Research (S)

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

Principal Investigator

SUTOH Kazuo  The University of Tokyo, 大学院・総合文化研究科, 教授 (20111453)

Co-Investigator(Kenkyū-buntansha) 栗栖 源嗣  東京大学, 大学院・総合文化研究科, 准教授 (90294131)
昆 隆英  東京大学, 大学院・総合文化研究科, 助教 (30332620)
大岩 和弘  情報通信研究機構, 生体物性グループ, リーダー (10211096)
Co-Investigator(Renkei-kenkyūsha) KON Takahide  東京大学, 大学院・総合文化研究科, 助教 (30332620)
Project Period (FY) 2005 – 2009
Project Status Completed (Fiscal Year 2009)
Budget Amount *help
¥107,250,000 (Direct Cost: ¥82,500,000、Indirect Cost: ¥24,750,000)
Fiscal Year 2009: ¥14,950,000 (Direct Cost: ¥11,500,000、Indirect Cost: ¥3,450,000)
Fiscal Year 2008: ¥17,030,000 (Direct Cost: ¥13,100,000、Indirect Cost: ¥3,930,000)
Fiscal Year 2007: ¥29,900,000 (Direct Cost: ¥23,000,000、Indirect Cost: ¥6,900,000)
Fiscal Year 2006: ¥34,060,000 (Direct Cost: ¥26,200,000、Indirect Cost: ¥7,860,000)
Fiscal Year 2005: ¥11,310,000 (Direct Cost: ¥8,700,000、Indirect Cost: ¥2,610,000)
Keywords細胞質ダイニン / AAA+ファミリー / 微小管 / ATP加水分解酵素 / FRET / 電子顕微鏡 / パワーストローク / ダイニン / ATP加水分解反応 / プロセシビティー / AAAリング / ストップトフロー / モータードメイン / ヘテロダイマータグ / モータータンパク質 / AAA+タンパク質 / クライオ電子顕微鏡法 / レバーアーム / ATP加水分解 / X線結晶解析 / 電子顕微鏡法 / 微小管滑り運動 / 蛍光エネルギー遷移(FRET) / GFP / アロステリック相互作用
Research Abstract

Our major goal of this project is to elucidate the molecular mechanism of energy transduction of cytoplasmic dynein by genetic engineering, structural studies and ATPase kinetics analyses. Our major results are: (1) we have identified the minimal motor domain of cytoplasmic dynein and examined its structure by electron microscopy. (2) Based on ATPase kinetics, we have identified three ATPase sites and one ADP-binding site in the minimal motor domain (380kDa), among which one is the primary ATPase site and the others are regulatory sites. (3) We have identified intermediate steps of the ATPase cycle by means of stopped-flow kinetics. (4) We have discovered that the linker protruding from the core ring swings as a lever-arm at specific steps of the ATPase cycle of the primary ATPase site. (5) We have shown that structural changes of the microtubule-binding domain of dynein are induced at specific ATPase steps of the primary ATPase site. Based on these findings, we have proposed a molecular model of energy transduction of cytoplasmic dynein.

Report

(6 results)
  • 2009 Annual Research Report   Final Research Report ( PDF )
  • 2008 Annual Research Report
  • 2007 Annual Research Report
  • 2006 Annual Research Report
  • 2005 Annual Research Report
  • Research Products

    (42 results)

All 2009 2008 2007 2006 2005 2004

All Journal Article (23 results) (of which Peer Reviewed: 7 results) Presentation (19 results)

  • [Journal Article] Biomolecular-Motor-Based Nano-or Microscale Particle Translocations on DNA Microarrays.2009

    • Author(s)
      Satoshi Hiyama,, Riho Gojo, Tomohiro Shima, Shoji Takeuchi, Kazuo Sutoh
    • Journal Title

      Nano Letters.Publication on Web on

    • Related Report
      2009 Final Research Report
  • [Journal Article] AAA+ Ring and Linker Swing Mechanism in the Dynein Motor2009

    • Author(s)
      Anthony J. Roberts, Naoki Numata, Matt L. Walker, Yusuke S. Kato, Bara Malkova, Takahide Kon, Reiko Ohkura, Fumio Arisaka, Peter J. Knight, Kazuo Sutoh, Stan A. Burgess.
    • Journal Title

      Cell 136

      Pages: 485-595

    • Related Report
      2009 Final Research Report
  • [Journal Article] Helix sliding in the stalk coiled coil of dynein couples ATPase and microtubule binding.2009

    • Author(s)
      Takahide Kon, Kenji Imamula, Anthony J Roberts, Reiko Ohkura, Peter J Knight, I R Gibbons, Stan A Burgess, Kazuo Sutoh
    • Journal Title

      Nature Structural and Molecular Biology 16

      Pages: 325-333

    • Related Report
      2009 Final Research Report
  • [Journal Article] AAA+ring and linker swing mechanism of the dynein motor2009

    • Author(s)
      Roberts, A., Numata, N., 等
    • Journal Title

      Cell 136

      Pages: 485-495

    • Related Report
      2009 Annual Research Report
    • Peer Reviewed
  • [Journal Article] Helix sliding of stalk coiled coil of dynein couples ATPase and microtubule bindin2009

    • Author(s)
      Kon, T., Imamula, K., 等
    • Journal Title

      Nature Structutal & Molecular Biology 16

      Pages: 325-333

    • Related Report
      2009 Annual Research Report
    • Peer Reviewed
  • [Journal Article] AAA+ Ring and Linker Swing Mechanism in the Dynein Motor2009

    • Author(s)
      Anthony J. Roberts, Kazuo Sutoh, Stan A. Burgess, et al.
    • Journal Title

      Cell 136

      Pages: 485-595

    • Related Report
      2008 Annual Research Report
    • Peer Reviewed
  • [Journal Article] Helix sliding in the stalk coiled coil of dynein couples ATP ase and microtubule bind2009

    • Author(s)
      Takahide Kon, Stan A Burgess, Kazuo Sutoh, et al.
    • Journal Title

      Nature Structural and Molecular Biology 16

      Pages: 325-333

    • Related Report
      2008 Annual Research Report
    • Peer Reviewed
  • [Journal Article] Autonomous Loading/Unloading and Transport of Specified Cargoes Using DNA Hybridization and Biological Motor-Based Motility.2008

    • Author(s)
      Hiyama, S., Inoue, T., Shima, T., Moritani, Y., Suda, T., Sutoh, K.
    • Journal Title

      Small 4

      Pages: 410-415

    • Related Report
      2009 Final Research Report
  • [Journal Article] Three-dimensional structure of cytoplasmic dynein bound to microtubules.2007

    • Author(s)
      Naoko Mizuno, Akihiro Narita, Takahide Kon, Kazuo Sutoh, Masahide Kikkawa
    • Journal Title

      PNAS 104

      Pages: 20832-20837

    • Related Report
      2009 Final Research Report
  • [Journal Article] The coordination of cyclic microtubule association/dissociation and tail swing of cytoplasmic dynein.2007

    • Author(s)
      Kenji Imamula, Takahide Kon, Reiko Ohkura, Kazuo Sutoh
    • Journal Title

      PNAS 104

      Pages: 16134-16134

    • Related Report
      2009 Final Research Report
  • [Journal Article] Kinetic characterization of tail swing steps in the ATPase cycle of Dictyostelium cytoplasmic dynein.2007

    • Author(s)
      Toshifumi Mogami, Takahide Kon*, Kohji Ito, Kazuo Sutoh
    • Journal Title

      J.Bio.Chem 282

      Pages: 21639-21639

    • Related Report
      2009 Final Research Report
  • [Journal Article] Three-dimensional structure of cytoplasmic dynein bound to microtubules2007

    • Author(s)
      Naoko Mizuno, Akihiro Narita, Takahide Kon, Kazuo Sutoh, and Masahide Kikkawa
    • Journal Title

      PNAS 104

      Pages: 20832-20837

    • Related Report
      2007 Annual Research Report
    • Peer Reviewed
  • [Journal Article] The coordination of cyclic microtubule association/dissociation and tail swing of cytoplasmic2007

    • Author(s)
      Kenji Imamula, Takahide Kon, Reiko Ohkura, and Kazuo Sutoh
    • Journal Title

      PNAS 104

      Pages: 16134-16139

    • Related Report
      2007 Annual Research Report
    • Peer Reviewed
  • [Journal Article] Kinetic characterization of tail swing steps in the ATPase cycle of Dictyostelium cytoplasmic2007

    • Author(s)
      Toshifumi Mogami, Takahide Kon, Kohji Ito, Kazuo Sutoh
    • Journal Title

      J.Bio.Chem 282

      Pages: 21639-21644

    • Related Report
      2007 Annual Research Report
    • Peer Reviewed
  • [Journal Article] Two modes of microtubule sliding driven by cytoplasmic dynein.2006

    • Author(s)
      Tomohiro Shima, Takahide Kon, Kenji Imamula, Reiko Ohkura, Kazuo Sutoh
    • Journal Title

      PNAS 103

      Pages: 17736-17740

    • Related Report
      2009 Final Research Report
  • [Journal Article] Functional characterization of the amino-terminal region of myosin 2.2006

    • Author(s)
      Setsuko Fujita-Becker, Georgios Tsiavaliaris, Reiko Ohkura, Takashi Shimada, Dietmar Manstein, Kazuo Sutoh
    • Journal Title

      J.Bio.Chem. 281

      Pages: 36102-36109

    • Related Report
      2009 Final Research Report
  • [Journal Article] Head-head coordination is required for the processive motion of cytoplasmic dynein, an AAA+ molecular motor.2006

    • Author(s)
      Tomohiro Shima, Kenji Imamula, Takahide Kon, Reiko Ohkura, Kazuo Sutoh
    • Journal Title

      J.Struc.Biol. 156

      Pages: 182-189

    • Related Report
      2009 Final Research Report
  • [Journal Article] Two modes of microtubule sliding driven by cytoplasmic dynein2006

    • Author(s)
      Tomohiro Shima, Takahide Kon, Kenji Imamula, Reiko Ohkura, Kazuo Sutoh
    • Journal Title

      PNAS 103

      Pages: 17736-17740

    • Related Report
      2006 Annual Research Report
  • [Journal Article] Head-head coordination is required for the processive motion of cytoplasmic dynein, an AAA+ molecular motor2006

    • Author(s)
      Tomohiro Shima, Kenji Imamula, Takahide Kon, Reiko Ohkura, Kazuo Sutoh
    • Journal Title

      J. Struc. Biol. 156

      Pages: 182-189

    • Related Report
      2006 Annual Research Report
  • [Journal Article] ATP-hydrolysis cycle-dependent stem motions in cytoplasmic dynein.2005

    • Author(s)
      Takahide Kon, Toshifumi Mogami, Reiko Ohkura, Masaya Nishiura, Kazuo Sutoh
    • Journal Title

      Nature Structural and Molecular Biology 12

      Pages: 513-519

    • Related Report
      2009 Final Research Report
  • [Journal Article] ATP-hydrolysis cycle-dependent stem motions in cytoplasmic dynein2005

    • Author(s)
      Takahide Kon, Toshifumi Mogami, Reiko Ohkura, Masaya Nishiura, Kazuo Sutoh
    • Journal Title

      Nature Structural and Molecular Biology 12

      Pages: 513-519

    • Related Report
      2005 Annual Research Report
  • [Journal Article] Distinct functions of nucleotide-binding/hydrolysis sites in the four AAA modules of cytoplasmic dynein, as revealed by biochemical characterizations of recombinant fragments.2004

    • Author(s)
      Takahide Kon, Masaya Nishiura, Reiko Ohkura, Yoko Y. Toyoshima, Kazuo Sutoh
    • Journal Title

      Biochemistry 43

      Pages: 11266-11274

    • Related Report
      2009 Final Research Report
  • [Journal Article] A single-headed recombinant fragment of Dictyostelium cytoplasmic dynein can drive the robust sliding of microtubules.2004

    • Author(s)
      Nishiura, M., Kon, T., Shiroguchi, K., Ohkura, R., Shima, T., Toyoshima, Y. Y., Sutoh, K.
    • Journal Title

      J.Biol.Chem. 279

      Pages: 22799-22802

    • Related Report
      2009 Final Research Report
  • [Presentation] Dynein-The old but still mysterious motor protein2009

    • Author(s)
      Sutoh K.
    • Organizer
      The Biophysical Society subgroup meeting
    • Related Report
      2009 Final Research Report
  • [Presentation] Elucidation of structural states of dimeric motor domain of dynein using Cys-light construct2009

    • Author(s)
      Hasegawa Y., Shima T., Kon T., Sutoh K., Sutoh K.
    • Organizer
      The Biophysical Society annual meeting
    • Place of Presentation
      USA
    • Related Report
      2009 Final Research Report
  • [Presentation] How does the two-headed dynein processively walk on a microtubule?2009

    • Author(s)
      Shima T., Numata N., Kon T., Ohkura R., Higuchi H., Sutoh K.
    • Organizer
      The Biophysical Society annual meeting
    • Place of Presentation
      USA
    • Related Report
      2009 Final Research Report
  • [Presentation] Communication of multiple ATPase sites of the dynein motor domain.2008

    • Author(s)
      Sutoh K.
    • Organizer
      Gordon Conference on Muscle and Molecular Motors
    • Related Report
      2009 Final Research Report
  • [Presentation] Dissection of Inter-modular Communication among Multiple Nucleotide-binding/hydrolysis Sites of Cytoplasmic Dynein.2008

    • Author(s)
      Mogami T., Imamula K., Ohkura R., Sutoh K., Kon T.
    • Organizer
      The American Society of Cell Biology annual meeting
    • Related Report
      2009 Final Research Report
  • [Presentation] Biochemical Dissection of Motor Functions of Cytoplasmic Dynein.2007

    • Author(s)
      Sutoh K.
    • Organizer
      The Biophysical Society annual meeting
    • Related Report
      2009 Final Research Report
  • [Presentation] Force generation by cytoplasmic dynein, a microtubule-based motor protein that belongs to the AAA+ family.2007

    • Author(s)
      Sutoh K.
    • Organizer
      The 6^<th> International Conference on AAA proteins
    • Related Report
      2009 Final Research Report
  • [Presentation] Kinetic analysis of dynein-MT interaction.2007

    • Author(s)
      Imamula K, Kon T, Ohkura R, Sutoh, K.
    • Organizer
      Biophysical Society annual meeting
    • Place of Presentation
      USA
    • Related Report
      2009 Final Research Report
  • [Presentation] Two modes of microtubule-sliding driven by cytoplasmic dynein.2007

    • Author(s)
      Shima T, Kon T, Imamula K, Ohkura R., Sutoh K.
    • Organizer
      The Biophysical Society annual meeting
    • Place of Presentation
      USA
    • Related Report
      2009 Final Research Report
  • [Presentation] Are six AAA modules sufficient to form the AAA ring of dynein?2007

    • Author(s)
      Numata N, Roberts A, Burgess SA, Kon, T., Ohkura, R., Knight, P. Sutoh, K.
    • Organizer
      The Biophysical Society annual meeting
    • Place of Presentation
      USA
    • Related Report
      2009 Final Research Report
  • [Presentation] Structure and Function of Dynein, A Huge Microtubule-based AAA+ Motor.2006

    • Author(s)
      Sutoh K.
    • Organizer
      The Biophysical Society annual meeting
    • Related Report
      2009 Final Research Report
  • [Presentation] Dissection of Intra-molecular Communications between the Catalytic Head and Microtubule-binding Site in the Dynein Heavy Chain.2006

    • Author(s)
      Kon T., Imamura K., Okura R., Sutoh K.
    • Organizer
      The American Society of Cell Biology annual meeting
    • Related Report
      2009 Final Research Report
  • [Presentation] Roles of multiple nucleotide-binding/hydrolysis sites of the cytoplasmic dynein motor domain in microtubule sliding2006

    • Author(s)
      Sutoh K., Mogami T., Imamula K., Ohkura R., Kon T.
    • Organizer
      The Biophysical Society Discussion Meeting
    • Place of Presentation
      USA
    • Related Report
      2009 Final Research Report
  • [Presentation] Dissection of Intra-molecular Communications between the Catalytic Head and Microtubule-binding Site in the Dynein Heavy Chain.2006

    • Author(s)
      Kon T., Imamura K., Okura R., Sutoh K.
    • Organizer
      The Biophysical Society Discussion Meeting
    • Place of Presentation
      USA
    • Related Report
      2009 Final Research Report
  • [Presentation] Molecular mechanism of force generation by cytoplasmic dynein as revealed by fully active recombinant motor domain.2005

    • Author(s)
      Sutoh K.
    • Organizer
      Gordon Conference on Muscle and Molecular Motors
    • Related Report
      2009 Final Research Report
  • [Presentation] Active participation of the stalk coiled-coil in communication between ATPase and MT-binding sites of cytoplasmic dynein2005

    • Author(s)
      Okura R, Kon T, Shima T., Sutoh K.
    • Organizer
      The Biophysical Society annual meeting
    • Place of Presentation
      USA
    • Related Report
      2009 Final Research Report
  • [Presentation] ATP-hydrolysis cycle dependent stem motions in cytoplasmic dynein.2005

    • Author(s)
      Kon T, Mogami T, Ohkura R, Sutoh K.
    • Organizer
      Biophysical Society annual meeting
    • Place of Presentation
      USA
    • Related Report
      2009 Final Research Report
  • [Presentation] Kinetic analysis of FRET changes of recombinant dynein motor domain.2005

    • Author(s)
      Mogami T, Kon T, Ohkura R, Sutoh K.
    • Organizer
      The Biophysical Society annual meeting
    • Place of Presentation
      USA
    • Related Report
      2009 Final Research Report
  • [Presentation] Distinct roles of the four nucleotide-binding/hydrolysis sites in the motor activities of cytoplasmic dynein.2004

    • Author(s)
      Kon T, Mogami T, Nishiura M, Sutoh K.
    • Organizer
      The American Society of Cell Biology annual meeting
    • Related Report
      2009 Final Research Report

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

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