High-resolution structural analysis of the molecular -motor-rail system

2008 Fiscal Year Final Research Report

• Project/Area Number: 16083102
• Research Category: Grant-in-Aid for Scientific Research on Priority Areas
• Allocation Type: Single-year Grants
• Review Section: Biological Sciences
• Research Institution: National Institute of Advanced Industrial Science and Technology
• Principal Investigator: HIROSE Keiko National Institute of Advanced Industrial Science and Technology, セルエンジニアリング研究部門, 主任研究員 (90357872)
• Co-Investigator(Kenkyū-buntansha): YASUNAGA Takuo 九州工業大学, 大学院情報工学研究院, 教授 (60251394)
• Project Period (FY): 2004 – 2008
• Keywords: 分子モーター / キネシン / ダイニン / ミオシン / 微小管 / 電子顕微鏡 / 構造 / 画像解析

Research Abstract

ダイニン・微小管複合体の電子顕微鏡構造研究を行い、従来のモデルに反して、力発生の前後に対応する状態でストークの角度が変化しないことを示し、新たなモデルを提唱した。キネシンファミリー分子Kar3と微小管の複合体の立体構造を約12Åの分解能で解き、ヌクレオチド状態に依存した構造変化を明らかにした。骨格筋ミオシンとアクチンの硬直複合体構造を1nmを超える分解能で得た。また、高速ミオシンのアクチン結合での構造変化を見出した

Research Products

• [Journal Article] ダイニン・微小管複合体の高分解能構造解析 (2009)
  • Author(s): 広瀬恵子、上野裕則
  • Journal Title: 顕微鏡 (in press)
  • Peer Reviewed
• [Journal Article] Expression of excess receptors and negative feedback control of signal pathways are required for rapid activation and prompt cessation of signal transduction (2009)
  • Author(s): Kobayashi, H., Azuma, R., and Yasunaga T.
  • Journal Title: Cell Communication and Signaling 7, 3
  • Peer Reviewed
• [Journal Article] Dynein pulls microtubules without rotating its stalk (2008)
  • Author(s): Ueno, H., Yasunaga, T., Shingyoji, C., and Hirose K.
  • Journal Title: Proc. Natl. Acad. Sci., USA. 105 Pages: 19702-19707
  • Peer Reviewed
• [Journal Article] Single-headed mode of kinesin-5 (2008)
  • Author(s): Kaseda, K., Crevel, I., Hirose, K. and Cross R.A.
  • Journal Title: EMBO Reports 9 Pages: 761-765
  • Peer Reviewed
• [Journal Article] Partially functional outer arm dynein in a novel Chlamydomonas mutant expressing a truncated γ heavy chain (2008)
  • Author(s): Liu Z., Takazaki H., Nakazawa Y., Sakato M., Yagi T., Yasunaga T., King S.M., and Kamiya R.
  • Journal Title: Eukaryotic Cell 7 Pages: 1136-1145
  • Peer Reviewed
• [Journal Article] Structural basis for tropomyosin overlap in thin (actin) filaments and the generation of a molecular swivel by troponin-T (2008)
  • Author(s): Murakami, K., Stewart, M., Nozawa, K., Tomii, K., Kudou, N., Igarashi, N., Shirakihara, Y., Wakatsuki, S., Yasunaga, T., and Wakabayashi, T.
  • Journal Title: Proc. Natl. Acad. Sci., USA. 105 Pages: 7200-7205
  • Peer Reviewed
• [Journal Article] Evaluation of a 2k CCD camera with an epitaxially-grown CsI scintillator for recording energy-filtered electron cryo-micrographs (2008)
  • Author(s): Yasunaga, T., Wakabayashi, T.
  • Journal Title: Journal of Electron Microscopy 57(3) Pages: 101-112
  • Peer Reviewed
• [Journal Article] A cool look at the structural changes in kinesin motor domains (2007)
  • Author(s): Hirose, K. and Amos, L. A.
  • Journal Title: J. Cell Science 120 Pages: 3919-3927
  • Peer Reviewed
• [Journal Article] A novel system for expressing toxic actin mutants in Dictyostelium and purification and characterization of a dominant lethal yeast actin mutant (2007)
  • Author(s): Noguchi, T.Q.P., Kanzaki, N., Ueno, H., Hirose, K. and Uyeda, T.
  • Journal Title: J. Bio. Chem. 282 Pages: 27721-27727
  • Peer Reviewed
• [Journal Article] A genetically encoded metallothioneintag enabling efficient protein detectionby electron microscopy (2007)
  • Author(s): Nishino, Y., Yasunaga, T. and Miyazawa, A.
  • Journal Title: Journal of Electron Microscopy 56(3) Pages: 93-101
  • Peer Reviewed
• [Journal Article] An actin homolog of the archaeon Thermoplasma acidophilum that retains the ancient trait of eukaryotic actin (2007)
  • Author(s): Hara, F., Yamashiro, K., Nemoto, N., Ohta, Y, Yokobori, S, Yasunaga, T., Hisanaga, S. and Yamagishi A.
  • Journal Title: J. Bacteriology 189(5) Pages: 2039-2045
  • Peer Reviewed
• [Journal Article] Domains 16 and 17 of tropoelastin in elastic fiber formation (2007)
  • Author(s): Wachi, H., Sato, F., Nakazawa, J., Nonaka, R., Szabo, Z., Urban, Z., Yasunaga, T., Maeda, I., Okamoto, K., Starcher, B.C., Li, D.Y., Mecham, R.P., and Seyama, Y.
  • Journal Title: Biochemical Journal 402 Pages: 60-70
  • Peer Reviewed
• [Journal Article] 電子顕微鏡画像解析ソフトウェア統合環境Eosの試み (2007)
  • Author(s): 安永卓生
  • Journal Title: 顕微鏡 42 Pages: 214-216
  • Peer Reviewed
• [Journal Article] Large Conformational Changes in a Kinesin Motor Catalysed by Interaction with Microtubules (2006)
  • Author(s): Hirose, K. Akimaru, E., Akiba, T., Endow, A. S., and Amos, L. A.
  • Journal Title: Mol. Cell 23(6) Pages: 913-923
  • Peer Reviewed
• [Journal Article] Comparative analysis of the protein folding activities of two chaperonin subunits of Thermococcus strain KS-1 ; the effects of beryllium fluoride (2006)
  • Author(s): Yoshida, T., Iizuka, R., Itami, K., Yasunaga, T., Sakuraba, H., Ohshima, T., Yohda, M., and Maruyama, T.
  • Journal Title: Extremophiles 11(2) Pages: 225-235
  • Peer Reviewed
• [Journal Article] Atomic model construction of protein complexes from electron micrographs and visualization of their 3D structure using VR system (2006)
  • Author(s): Noda, K., Nakamura, M., Nishida, R., Yamaguchi, Y., Tamura, Y., Nakamura, H., and Yasunaga, T.
  • Journal Title: J. Plasma Phys. 72(06) Pages: 1037-1040
  • Peer Reviewed
• [Journal Article] A bidirectional Kinesin motor in live Drosophila embryos (2005)
  • Author(s): Sciambi, C. J., Komma, D. J., Skold, H. N., Hirose, K., Endow, S. A.
  • Journal Title: Traffic 6(11) Pages: 1036-1046
  • Peer Reviewed
• [Journal Article] An axonemal dynein particularly important for flagellar movement at high viscosity : Implications from a new chlamydomonas mutant deficient in the dynein heavy chain gene DHC9 (2005)
  • Author(s): Yagi T., Minoura I, Fujiwara A., Saito R., Yasunaga T., Hirono M., and Kamiya R.
  • Journal Title: J. Biol. Chem. 280 Pages: 41412-41420
  • Peer Reviewed
• [Journal Article] Structural basis for Ca^<2+>-regulated muscle relaxation at interaction sites of troponin with actin and tropomyosin (2005)
  • Author(s): Murakami, K, Yumoto, F, Ohki, SY, Yasunaga, T, Tanokura, M, and Wakabayashi, T.
  • Journal Title: J. Mol. Biol. 352 Pages: 178-201
  • Peer Reviewed
• [Journal Article] Automatic detecting particles approach using adaboost-SVM (2005)
  • Author(s): Yang, J., Ohashi, T., and Yasunaga, T.
  • Journal Title: Wseas Transactinns on Biology and Biomedicine 2 Pages: 163-168
  • Peer Reviewed
• [Presentation] クライオ電子顕微鏡法の現状と将来 (2008)
  • Author(s): 安永卓生
  • Organizer: 日本顕微鏡学会九州支部総会・学術講演会
  • Place of Presentation: 久留米市, 招待講演.
  • Year and Date: 20081200
• [Presentation] What cryo-EM images tell us about the mechanism of dynein motility. (2008)
  • Author(s): 広瀬恵子
  • Organizer: 日本生物物理学会第46回年会
  • Place of Presentation: 福岡, 依頼講演.
  • Year and Date: 2008-12-04
• [Presentation] Cryo-EM studies of Microtubule Motors. (2008)
  • Author(s): Hirose, K.
  • Organizer: Gordon Research Conferences on Muscle & Molecular Motors.
  • Place of Presentation: New London, NH, USA, 招待講演
  • Year and Date: 2008-06-29
• [Presentation] タンパク質の可視化技術の最前線 (2007)
  • Author(s): 安永卓生
  • Organizer: 市民フォーラム「生命科学の進歩とバイオマス利用最前線」、主催 日本生物工学会九州支部
  • Place of Presentation: 依頼講演.
  • Year and Date: 2007-08-10
• [Presentation] 微小管に結合した分子モーターキネシンおよびダイニンの構造解析. (2006)
  • Author(s): 広瀬恵子
  • Organizer: 日本顕微鏡学会生体構造解析分科会研究討論会
  • Place of Presentation: 大阪, 招待講演
  • Year and Date: 2006-12-21
• [Presentation] 生体内モータータンパク質の電子顕微鏡法による3次元構造解析 (2005)
  • Author(s): 安永卓生, 齊藤良, 太田好則, 狩野要介, 若林健之
  • Organizer: 第50回日本顕微鏡学会シンポジウム
  • Place of Presentation: 招待講演
  • Year and Date: 2005-11-01
• [Presentation] 分子モーターのメカニズム解明に向けた電子顕微鏡による構造解析 (2005)
  • Author(s): 安永卓生
  • Organizer: 第29回蛋白質と酵素の構造と機能に関する九州シンポジウム
  • Place of Presentation: 招待講演
  • Year and Date: 2005-09-15
• [Presentation] Nucleotide-Dependent Structural Changes in Kar3 Motor Domains Complexed to MTs. (2004)
  • Author(s): Hirose, K.
  • Organizer: The American Society for Cell Biology, 44th Annual Meeting
  • Place of Presentation: Washington, USA, 依頼講演.
  • Year and Date: 2004-12-04
• [Book] 「ナノイメージング」第一編ナノイメージングを可能にする顕微鏡法・第3章電子顕微鏡を用いたナノイメージング. P.44-57 (2008)
  • Author(s): 安永卓生(株)NTS
• [Book] Methods Mol. Biol. 392巻, 213-230 (2007)
  • Author(s): Hirose, K. and Amos, L. A.
  • Publisher: Humana Press (USA)
• [Book] Methods Mol. Med. 137 巻, 65-91 (2007)
  • Author(s): Amos, L. A. and Hirose K.
  • Publisher: Humana Press (USA)
• [Book] Adv Exp Med Biol. 592, 71-86 (2007). Review. (2007)
  • Author(s): Murakami K, Yumoto F, Ohki SY, Yasunaga T, Tanokura M, Wakabayashi T.
  • Publisher: Structural basis for calcium-regulated relaxation of striated muscles at interaction sites of troponin with actin and tropomyosin.
• [Book] 実験化学講座 日本化学会・編 共著 電子顕微鏡の節 p642-650 (2006)
  • Author(s): 安永卓生
  • Publisher: 丸善
• [Remarks] プレス発表 : 分子モーターのメカニズムを解明 -ダイニン分子モーターの「腕」を可視化. 広瀬恵子、上野裕則、安永卓生、真行寺千佳子, 2008 年12 月2 日, (東大理学部)