Mechanical Regulation of cytoplasmic dynein motility

• Project/Area Number: 25291031
• Research Category: Grant-in-Aid for Scientific Research (B)
• Allocation Type: Partial Multi-year Fund
• Section: 一般
• Research Field: Biophysics
• Research Institution: The University of Tokyo
• Principal Investigator: TOYOSHIMA Yoko 東京大学, 総合文化研究科, 教授 (40158043)
• Co-Investigator(Kenkyū-buntansha): YAJIMA Junichiro 東京大学, 大学院総合文化研究科, 准教授 (00453499)
• Research Collaborator: KOBAYASHI Takuya ; SUGAWA Mitsuhiro ; TORISAWA Takayuki ; ICHIKAWA Munenori ; SAITO Kei ; KAJITA Hatsuha ; MIYASHITA Takuya ; SHIBATA Keitaro ; UTSUMI Yui ; FURUTA Ken'ya
• Project Period (FY): 2013-04-01 – 2016-03-31
• Project Status: Completed (Fiscal Year 2015)
• Budget Amount: ¥17,810,000 (Direct Cost: ¥13,700,000、Indirect Cost: ¥4,110,000); Fiscal Year 2015: ¥3,640,000 (Direct Cost: ¥2,800,000、Indirect Cost: ¥840,000); Fiscal Year 2014: ¥5,850,000 (Direct Cost: ¥4,500,000、Indirect Cost: ¥1,350,000); Fiscal Year 2013: ¥8,320,000 (Direct Cost: ¥6,400,000、Indirect Cost: ¥1,920,000)
• Keywords: 細胞質ダイニン / 微小管 / モーター活性 / 頭部間距離 / メカニカル制御 / ブラウン運動 / 方向性の運動 / キネシン / ダイナクチン / ダイニン / 自己制御機構 / 拡散運動 / 一方向運動 / スタック構造 / 1分子運動観察 / 電子顕微鏡観察 / ラチェット機構 / キャッチ機構

Outline of Final Research Achievements

To investigate the molecular mechanism of dynein motility, we have genetically engineered human dynein heavy chain and observed single molecule behaviors of the recombinant proteins on the microtubule and their molecular shape by electron microscopy. When dynein didn’t have the cargo, namely under no load, the ring structure of the two heads overlapped in face-to-face fashion and the dynein molecules fluctuated on the microtubule without showing unidirectional movement, falling in self-inhibited state. When the two heads were restricted not to overlapped, the molecules exhibited the unidirectional and processive movement toward the microtubule minus end under no load.

Research Products

• [Journal Article] Axonemal dynein light chain-1 locates at the microtubule-binding domain of the γ heavy chain. (2015)
  • Author(s): Ichikawa M, Saito K, Yanagisawa HA, Yagi T, Kamiya R, Yamaguchi S, Yajima J, Kushida Y, Nakano K, Numata O and Toyoshima YY.
  • Journal Title: Molicular Biology of the Cell Volume: 26 Issue: 23 Pages: 4236-47
  • DOI: 10.1091/mbc.e15-05-0289
  • Peer Reviewed / Open Access
• [Journal Article] α- and β-tubulin lattice arrangement of the axonemal microtubule doublet and binding proteins revealed by single particle cryo-electron microscopy and tomography. (2015)
  • Author(s): Maheshwari, A., Obbineni, J. M., Bui, K. H., Shibata, K., Toyoshima Y. Y. & Ishikawa, T.
  • Journal Title: Structure Volume: 23 Issue: 9 Pages: 1584-1595
  • DOI: 10.1016/j.str.2015.06.017
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Regulatory Mechanism of Cytoplasmic Dynein (2015)
  • Author(s): 鳥澤嵩征、古田健也、豊島陽子
  • Journal Title: Seibutsu Butsuri Volume: 55 Issue: 3 Pages: 127-132
  • DOI: 10.2142/biophys.55.127
  • NAID: 130005072019
  • ISSN: 0582-4052, 1347-4219
  • Peer Reviewed / Open Access
• [Journal Article] Autoinhibition and cooperative activation mechanisms of cytoplasmic dynein (2014)
  • Author(s): Torisawa, T., Ichikawa, M., Furuta, A., Saito, K., Oiwa, K., Kojima, H., Toyoshima, Y. Y. and Furuta, K.
  • Journal Title: Nature Cell Biol Volume: 16 Issue: 11 Pages: 1118-1118
  • DOI: 10.1038/ncb3048
  • Peer Reviewed / Acknowledgement Compliant
• [Journal Article] Structure of the entire stalk region of the dynein motor domain (2014)
  • Author(s): Yosuke Nishikawa, Takuji Oyama, Narutoshi Kamiya, Takahide Kon, Yoko Y. Toyoshima, Haruki Nakamura, Genji Kurisu
  • Journal Title: Journal of Molecular Biology Volume: 426 Issue: 19 Pages: 3232-3245
  • DOI: 10.1016/j.jmb.2014.06.023
  • Peer Reviewed
• [Journal Article] バイオイメージングと光ピンセットを用いた微小管系モータータンパク質の協働的活性化に関する働き (2014)
  • Author(s): 古田健也、鳥澤嵩征、豊島陽子
  • Journal Title: 生化学 Volume: 86 Pages: 184-191
  • Peer Reviewed
• [Journal Article] Formation of ring-shaped microtubule assemblies through active self-organization on dynein. (2014)
  • Author(s): Ito, M., Arif Md. Rashedul Kabir, A.,M.,R. Inoue, D., Torisawa, T, Toyoshima, Y.Y., Sada1, K. & Kakugo
  • Journal Title: Polymer Jouranl Volume: 46 Pages: 220-225
  • NAID: 120005515308
  • Peer Reviewed
• [Journal Article] Sea lily muscle lacks a troponin-regulatory system but contains paramyosin. (2014)
  • Author(s): Obinata T, Amemiya S, Takai R, Ichikawa M, Toyosima YY, Sato N
  • Journal Title: Zoological Science Volume: 31 Issue: 3 Pages: 122-128
  • DOI: 10.2108/zsj.31.122
  • Peer Reviewed
• [Journal Article] Measuring Collective Transport by Defined Numbers of Processive and Nonprocessive Kinesin Motors (2013)
  • Author(s): 古田 健也, 古田 茜, 豊島 陽子, 網野 美紗子, 大岩 和弘, 小嶋 寛明
  • Journal Title: Proc Natl Acad Sci U S A ( 米国科学アカデミー紀要) Volume: 110(2) Issue: 2 Pages: 501-6
  • DOI: 10.1073/pnas.1201390110
  • Peer Reviewed
• [Journal Article] Formation of Ring-Shaped Microtubule Assemblies through Active Self-Organization on Dynein (2013)
  • Author(s): Masaki Ito, Arif Md Rashedul Kabir, Daisuke Inoue, Takayuki Torisawa, Yoko Toyoshima, Kazuki Sada, Akira Kakugo
  • Journal Title: Polymer Journal Volume: 46 Issue: 4 Pages: 220-225
  • DOI: 10.1038/pj.2013.89
  • NAID: 120005515308
  • Peer Reviewed
• [Presentation] Axonemal Dynein Light Chain-1 Locates at the Microtubule Binding Domain of the γ Heavy Chain (2015)
  • Author(s): Ichikawa M., Saito K., Yanagisawa H., Yagi, T., Kamiya, R., Yamaguchi S., Yajima, J., Kushida Y., Nakano, K., Numata, O. and Toyoshima, Y. Y.
  • Organizer: 第５３回日本生物物理学会年会
  • Place of Presentation: 金沢大学（石川県・金沢市）
  • Year and Date: 2015-09-13
• [Presentation] 分裂期kinesin-6の運動特性に関する研究 (2015)
  • Author(s): Yohei Maruyama, Akihiko Sato, Tim Davis, Toshihisa Osaki, Shin Yamaguchi, Shoji Takeuchi, Masanori Mishima, Junichiro Yajima
  • Organizer: 第５３回日本生物物理学会年会
  • Place of Presentation: 金沢大学（石川県・金沢市）
  • Year and Date: 2015-09-13
• [Presentation] Off-axis motion of yeast cytoplasmic dynein takes a biased random walk (2015)
  • Author(s): Sugawa, M., Yamaguchi, S., Takagi, H., Shibata, K., Toyoshima, Y. Y., Yajima, J
  • Organizer: 第５３回日本生物物理学会年会
  • Place of Presentation: 金沢大学（石川県・金沢市）
  • Year and Date: 2015-09-13
• [Presentation] Dynactin is a biphasic regulator to promote and inhibit dynein motility (2015)
  • Author(s): Kobayashi, K., Miyashita, K., Kajita, H., Saito, K., Murayama, T., Toyoshima, Y. Y.
  • Organizer: 第５３回日本生物物理学会年会
  • Place of Presentation: 金沢大学（石川県・金沢市）
  • Year and Date: 2015-09-13
• [Presentation] 如何にしてキネシンは運動方向を逆にするのか (2015)
  • Author(s): 山岸雅彦、矢島潤一郎
  • Organizer: 第5回分子モーター討論会
  • Place of Presentation: 東京大学（東京・目黒区）
  • Year and Date: 2015-06-13
  • Invited
• [Presentation] Cytoplasmic dynein takes a route switching randomly between protofilaments with a bias toward the right (2014)
  • Author(s): Mitsuhiro Sugawa, Shin Yamaguchi, Keitaro Shibata, Yoko Y. Toyoshima, Junichiro Yajima
  • Organizer: 第52回日本生物物理学会年会
  • Place of Presentation: 札幌コンベンションセンター（北海道）
  • Year and Date: 2014-09-13 – 2014-09-15
• [Presentation] Autoinhibiton and cooperative activation mechanisms of cytoplasmic dynein. (2014)
  • Author(s): Takayuki Torisawa, Ken’ya Furuta, Muneyoshi Ichikawa, Akane Furuta, Kei Saito, Kazuhiro Oiwa, Hiroaki Kojima, Yoko Toyoshim
  • Organizer: 第52回日本生物物理学会年会
  • Place of Presentation: 札幌コンベンションセンター（北海道）
  • Year and Date: 2014-09-13 – 2014-09-15
• [Presentation] A novel role of dynactin for dynein motility revealed by gliding assay (2014)
  • Author(s): Takuya Kobayashi, Hatsuha Kajita, Kei Saito, Yoko Y. Toyoshima
  • Organizer: 第52回日本生物物理学会年会
  • Place of Presentation: 札幌コンベンションセンター（北海道）
  • Year and Date: 2014-09-13 – 2014-09-15
• [Presentation] A novel role of dynactin for dynein motility revealed by single-molecule assay (2014)
  • Author(s): Takuya Miyashita, Takuya Kobayashi, Hatsuha Kajita, Yoko Y. Toyoshima
  • Organizer: 第52回日本生物物理学会年会
  • Place of Presentation: 札幌コンベンションセンター（北海道）
  • Year and Date: 2014-09-13 – 2014-09-15
• [Presentation] New Folding Pattern of p150 and “antenna” for dynein binding (2014)
  • Author(s): Kei Saito, Takashi Murayama, Tomonori Hata, Takuya Kobayashi, Yoko Toyoshima
  • Organizer: Biophysical Society 58th Annual Meeting
  • Place of Presentation: San Francisco, USA
• [Presentation] Self-regulation of cytoplasmic dynein through its unconventional force response. (2014)
  • Author(s): Takayuki Torisawa, Ken’ya Furuta, Akane Furuta, Muneyoshi Ichikawa, Kei Saito, Kazuhiro Oiwa, Hiroaki Kojima, Yoko Toyoshima
  • Organizer: Biophysical Society 58th Annual Meeting
  • Place of Presentation: San Francisco, USA
• [Presentation] Torque generation of axonemal outer arm dyneins (2014)
  • Author(s): Shin Yamaguchi, Kei Saito, Miki Sutoh, Takayuki Nishizaka, Yoko Y Toyoshima, Junichiro Yajima,
  • Organizer: Biophysical Society 58th Annual Meeting
  • Place of Presentation: San Francisco, USA
• [Presentation] Autoinhibition and synergistid activation of cytoplasmic dynein (2013)
  • Author(s): Takayuki Torisawa, Ken’ya Furuta, Akane Furuta, Muneyoshi Ichikawa, Yoko Toyoshima
  • Organizer: 日本生物物理学会第51回年会
  • Place of Presentation: 京都府、京都国際会議場
• [Presentation] Regulatory mechanism of mictrotubule –based molecular motors by Bicaudal-D2 (2013)
  • Author(s): Takuya Kobayashi, Akira Hanashima, Yoko Toyoshima, Takashi Murayama
  • Organizer: 日本生物物理学会第51回年会
  • Place of Presentation: 京都府、京都国際会議場
• [Presentation] Molecular architecture of dynactin p150 (2013)
  • Author(s): Kei Saito, Takashi Murayama, Tomonori Hata, Yoko Toyoshima
  • Organizer: 日本生物物理学会第51回年会
  • Place of Presentation: 京都府、京都国際会議場
• [Presentation] Subunit structure of Tetrahymena outer dynein arm complex (2013)
  • Author(s): Muneyoshi Ichikawa, Yasuharu Kushida, Kentaro Nakano, Osamu Numata, Yoko Toyoshima
  • Organizer: 日本生物物理学会第51回年会
  • Place of Presentation: 京都府、京都国際会議場
• [Presentation] The mechanism of determining the directionality of Ncd (2013)
  • Author(s): Masahiko Yamagishi, Yoko Toyoshima, Junichiro Yajima
  • Organizer: 日本生物物理学会第51回年会
  • Place of Presentation: 京都府、京都国際会議場
• [Presentation] Submolecular structure of cytoplasmic dynein tail and dynactin sidearm. (2013)
  • Author(s): Kei Saito, Yuta Watahabe, Muneyoshi Ichikawa, Tomonori Hata, Takuya Kobayashi, Takashi Murayama, Yoko Toyoshima
  • Organizer: International Workshop Dynein2013
  • Place of Presentation: 兵庫県、神戸オルビスホール
  • Invited
• [Presentation] Self-regulation of cytoplasmic dynein (2013)
  • Author(s): Ken’ya Furuta, Takayuyki Torisawa, Akane Furuta, Muneyoshi Ichikawa, Kei Saito, Kazuhiro Oiwa, Hiroaki Kojima, Yoko Toyoshima
  • Organizer: International Workshop Dynein2013
  • Place of Presentation: 兵庫県、神戸オルビスホール
  • Invited
• [Presentation] Torque generation by axonemal outer-arm dynein (2013)
  • Author(s): Shin Yamaguchi, Yoko Toyoshima, Junichiro Yajima
  • Organizer: International Workshop Dynein2013
  • Place of Presentation: 兵庫県、神戸オルビスホール
  • Invited
• [Presentation] Self-regulation of cytoplasmic dynein (2013)
  • Author(s): Ken'ya Furuta, Takayuki Torisawa, Akane Furuta, Muneyoshi Ichikawa, Kei Saito, Kauzuhiro Oiwa, Hiroaki Kojima, Yoko Toyoshima
  • Organizer: American Society for Cell Biology 2013 Annual Meeting
  • Place of Presentation: New Orleans, USA
• [Presentation] Submolecular structure of dynactin sidearm: a new folding pattern of p150 (2013)
  • Author(s): Kei Saito, Tomonori Hata, Takuya Kobayashi, Takashi Murayama, Yoko Toyoshima
  • Organizer: American Society for Cell Biology 2013 Annual Meeting
  • Place of Presentation: New Orleans, USA