Regulation of information flow by motor protein systems

2008 Fiscal Year Final Research Report

• Project/Area Number: 16083205
• Research Category: Grant-in-Aid for Scientific Research on Priority Areas
• Allocation Type: Single-year Grants
• Review Section: Biological Sciences
• Research Institution: The 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(Renkei-kenkyūsha): KON Takahide 東京大学, 大学院・総合文化研究科, 助教 (30332620)
• Project Period (FY): 2004 – 2008
• Keywords: モーターたんぱく質 / ミオシン / ダイニン / AAA+ATPase / FRET

Research Abstract

Our major goal of this project is to elucidate structures and functions of motor proteins, especially, unconventional myosin 6 and cytoplasmic dynein. We carried out the yeast two-hybrid screening of interacting partners of myosin 6. We identified several proteins including a membrane-bound tyrosine kinase. We examined in vivo functions of the kinase by using various cell- and molecular-biological techniques such as RNAi. We also examined structure and function of cytoplasmic dynein, using recombinant dynein motor domain. We identified interesting biochemical and structural properties of the motor domain by electron microscopy, rapid kinetics and FRET (fluorescence resonance energy transfer). The data obtained by these experiments will become the basis for understanding cellular functions of motor proteins.

Research Products

• [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
• [Journal Article] AAA+ Ring and Linker Swing Mechanism in the Dynein Motor (2009)
  • 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
• [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
• [Journal Article] BREK/LMTK2 is a myosin VI-binding protein involved in endosomal membrane trafficking (2008)
  • Author(s): Takeshi Inoue, Takahide Kon, Reiko Ohkura, Hisashi Yamakawa, Osamu Ohara, Jun Yokota, Kazuo Sutoh
  • Journal Title: Genes to Cells 13 Pages: 483-495
• [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
• [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
• [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
• [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
• [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
• [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
• [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
• [Journal Article] MYO18B interacts with the proteasomal subunit Sug1 and is degraded by the ubiquitin-proteasome pathway. (2006)
  • Author(s): Takeshi Inoue, Takahide Kon, Rieko Ajima, Reiko Ohkura, Masachika Tani, Jun Yokota, Kazuo Sutoh
  • Journal Title: Biochemical and Biophysical Research Communications 342 Pages: 829-834
• [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
• [Journal Article] The N-terminal extension of MYO18A has a ATP-insensitive actin-binding site (2005)
  • Author(s): Yasushi Isogawa, Takahide Kon, Takeshi Inoue, Reiko Ohkura, Hisashi Yamakawa, Osamu Ohara, Kazuo Sutoh
  • Journal Title: Biochemstry 44 Pages: 6190-6196
• [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
• [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
• [Presentation] Dynein-The old but still mysterious motor protein (2009)
  • Author(s): Sutoh K
  • Organizer: The Biophysical Society subgroup meeting
  • Year and Date: 20090000
• [Presentation] Elucidation of structural states of dimeric motor domain of dynein using Cys-light construct (2009)
  • Author(s): Hasegawa Y., Shima T., Kon T., Sutoh K., Sutoh K
  • Organizer: The Biophysical Society annual meeting
  • Place of Presentation: USA
  • Year and Date: 20090000
• [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: he Biophysical Society annual meeting
  • Place of Presentation: USA
  • Year and Date: 20090000
• [Presentation] Communication of multiple ATPase sites of the dynein motor domain (2008)
  • Author(s): Sutoh K
  • Organizer: Gordon Conference on Muscle and Molecular Motors
  • Year and Date: 20080000
• [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
  • Year and Date: 20080000
• [Presentation] Biochemical Dissection of Motor Functions of Cytoplasmic Dynein (2007)
  • Author(s): Sutoh K
  • Organizer: The Biophysical Society annual meeting
  • Year and Date: 20070000
• [Presentation] Force generation by cytoplasmic dynein, a microtubule-based motor protein that belongs to the AAA+ family (2007)
  • Author(s): Sutoh K
  • Organizer: The 6th International Conference on AAA proteins
  • Year and Date: 20070000
• [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
  • Year and Date: 20070000
• [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
  • Year and Date: 20070000
• [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
  • Year and Date: 20070000
• [Presentation] Structure and Function of Dynein, A Huge Microtubule-based AAA+ Motor (2006)
  • Author(s): Sutoh K
  • Organizer: The Biophysical Society annual meeting
  • Year and Date: 20060000
• [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
  • Year and Date: 20060000
• [Presentation] Roles of multiple nucleotide-binding/hydrolysis sites of the cytoplasmic dynein motor domain in microtubule sliding (2006)
  • Author(s): Sutoh K., Mogami T., Imamula K., Ohkura R., Kon T
  • Organizer: The Biophysical Society Discussion Meeting
  • Place of Presentation: USA
  • Year and Date: 20060000
• [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
  • Year and Date: 20060000
• [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
  • Year and Date: 20050000
• [Presentation] Interaction of myosin VI with a transmembrane Ser/Thr kinase KPI-2 (2005)
  • Author(s): Inoue T, Kon T, Ohkura R, Sutoh K
  • Organizer: The Biophysical Society annual meeting
  • Place of Presentation: USA
  • Year and Date: 20050000
• [Presentation] Active participation of the stalk coiled-coil in communication between ATPase and MT-binding sites of cytoplasmic dynein (2005)
  • Author(s): Okura R, Kon T, Shima T., Sutoh K
  • Organizer: The Biophysical Society annual meeting
  • Place of Presentation: USA
  • Year and Date: 20050000
• [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
  • Year and Date: 20050000
• [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
  • Year and Date: 20050000
• [Presentation] Interaction of myosin VI with a transmembrane Ser/Thr kinase KPI-2 (2004)
  • Author(s): Inoue T, Kon T, Ohkura R, Sutoh K. Cell Biology
  • Organizer: The American Society of Cell Biology annual meeting
  • Year and Date: 20040000
• [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
  • Year and Date: 20040000