電子顕微鏡法によるアクチンフィラメント重合核形成機構の解明

2011 Fiscal Year Annual Research Report

• Project/Area Number: 22770145
• Research Institution: Nagoya University
• Principal Investigator: 成田 哲博 名古屋大学, 理学研究科, 助教 (30360613)
• Keywords: アクチン / 電子顕微鏡 / 三次元構造 / 細胞骨格 / フィラメント極性 / 単粒子解析

Research Abstract

私たちは、アクチンフィラメント重合核形成機構の解明のため、本研究において、
Arp2/3複合体とフォルミンの研究を進めた。Arp2/3複合体は、細胞内において、膜近傍のアクチンフィラメントB端(重合端)を枝分かれさせ、新しいB端を増やすという形で重合核を形成する。しかし、その細胞内の正確な分布はわかっておらず、枝分かれ構造の詳細な三次元構造解析も行われていなかった。私たちはオーストリアのJohn Victor Small研究室と共同で、電子線トモグラフィーと負染色法を組み合わせた新しい手法を用いることで、細胞内のアクチンフィラメントネットワークの構造解析に成功。葉状仮足内の各アクチンフィラメントの極性、Arp2/3複合体が作る枝分かれ構造分布をミオシンや抗体等のラベルなしに、純粋に構造情報だけから決定することができたJ.Mol.Biol,2012, J. Cell. Sci, 2012)。本手法は広く細胞骨格の研究に非常に有用であると期待している。また、Arp2/3複合体を含んだ枝分かれ構造の平均化も進めており、現在分解能は2.9nm程度であるが、分解能2nm程度を目指してトモグラフィーの条件決定を進めている。

Research Products

• [Journal Article] Direct determination of actin polarity in the cell (2012)
  • Author(s): Narita A., Muekker J., Urban E., Vinzenz M., Small V., Maeda Y
  • Journal Title: J.Mol.Biol Volume: (印刷中)(in press)
  • DOI: 10.1016/j.jmb.2012.03.015
  • Peer Reviewed
• [Journal Article] Actin Branching in the initiation and maintenance of lamellipodia (2012)
  • Author(s): Vinzenz M Nemethova M., Schur F., Mueller J Narita A., Urban E., Winkler C., Koestler S.A., Rottner K., Resch G.P., Maeda Y., Small V.
  • Journal Title: J.Cell Sci. Volume: (印刷中)(in press)
  • DOI: 10.1242/jcs.107623
  • Peer Reviewed
• [Journal Article] Minimum requirements for the actin-like treadmilling motor system (2011)
  • Author(s): Narita A.
  • Journal Title: BioArchitecture Volume: 1(5) Pages: 205-208
  • DOI: 10.4161/bioa.1.5.18115
  • Peer Reviewed
• [Journal Article] Merits of the double-stranded form of the actin filament revealed by structures of the filament ends (2011)
  • Author(s): Narita A.
  • Journal Title: Communicative and Integrative Biology Volume: 4(6) Pages: 692-695
  • DOI: http://www.ncbi.nlm.nih.gov/pubmed/22446529
  • Peer Reviewed
• [Presentation] 分子と細胞をつなぐ線維状蛋白質構造解析 (2012)
  • Author(s): 成田哲博
  • Organizer: 第117回日本解剖学会総会・全国学術集会
  • Place of Presentation: 山梨大学(甲府市)(招待講演)
  • Year and Date: 2012-03-28
• [Presentation] Structural analysis of the actin filament in vitro and in vivo (2012)
  • Author(s): Narita A.
  • Organizer: International Workshop "from Structure to Dynamics" for Our Understanding of Protein-Protein Interactions
  • Place of Presentation: 名古屋大学(名古屋市)
  • Year and Date: 2012-03-17
• [Presentation] Spatial structure of cytoskeleton associated with nuclear membrane (2012)
  • Author(s): MinakataS., Koike M., Njirita A., Maeda Y., Usukura J.
  • Organizer: International Workshop "from Structure to Dynamics" for Our Understanding of Protein-Protein Interactions
  • Place of Presentation: 名古屋大学(名古屋市)
  • Year and Date: 2012-03-16
• [Presentation] Supramolecular structural analysis by STEM electron tomography (2012)
  • Author(s): Koike M., Minakata S.Narita A., Homma M., Usukura J., Maeda Y.
  • Organizer: International Workshop "from Structure to Dynamics" for Our Understanding of Protein-Protein Interactions
  • Place of Presentation: 名古屋大学(名古屋市)
  • Year and Date: 2012-03-16
• [Presentation] Two distinct mechanisms for actin capping protein regulation (2012)
  • Author(s): Takeda S., Minakata S., Koike R., Kawahata I., Narita A., Kitazawa M., Ota M., Yamakuni T., Maeda Y., Nitanai Y.
  • Organizer: International Workshop "from Structure to Dynamics" for Our Understanding of Protein-Protein Interactions
  • Place of Presentation: 名古屋大学(名古屋市)
  • Year and Date: 2012-03-16
• [Presentation] New method for identify the ABP binding sites on the actiti filament (2012)
  • Author(s): Dai S., Natita A., Ito T., Akaoka K., Maeda Y.
  • Organizer: International Workshop "from Structure to Dynamics" for Our Understanding of Protein-Protein Interactions
  • Place of Presentation: 名古屋大学(名古屋市)
  • Year and Date: 2012-03-16
• [Presentation] Actin Polymerization correlates with the flattening of actin molecule (2012)
  • Author(s): Iwasa M., Maeda K., Narita A., Aihara T., Maeda Y., Oda T.
  • Organizer: 56th Annual meeting Biophysical Society
  • Place of Presentation: San Diego Convention Center (San Diego)
  • Year and Date: 2012-02-27
• [Presentation] Structural analysis of the actin filament network in the cell (2012)
  • Author(s): Narita A.
  • Organizer: Direction of future in biological electron microscopy
  • Place of Presentation: 東京大学(文京区)(招待講演)
  • Year and Date: 2012-02-17
• [Presentation] Structural analysis of Na^+driven flagellar motor embedded in the membrane of Vibrio alginolyticus by electron tomography (2012)
  • Author(s): Koike M., Narita A., Homma M., Maeda Y.
  • Organizer: Sensory Transduction in Microorganisms at Grordon Research Conference
  • Place of Presentation: Ventura Beach Marriott(Ventura)
  • Year and Date: 2012-01-18
• [Presentation] Structural analysis of the actin filament in vivo and in vitro (2012)
  • Author(s): Narita A.
  • Organizer: Joint Seminar Centre for BioImaging Sciences-Structural biologydivision of Japan EM
  • Place of Presentation: シンガポール国立大学(Singapore)(招待講演)
  • Year and Date: 2012-01-12
• [Presentation] 分子から細胞までをつなぐ繊維状蛋白質複合体構造解析 (2011)
  • Author(s): 成田哲博
  • Organizer: 第2回神経科学と構造生物学の融合
  • Place of Presentation: 岡崎コンファレンスセンター(岡崎市)(招待講演)
  • Year and Date: 2011-11-22
• [Presentation] Two distinct mechanisms for actin capping protein regulation (2011)
  • Author(s): Takeda S., Minakata S., Koike R., Kawahata I., Narita A., Kitazawa M., Ota M., Yamakuni T., Maeda Y., Nitanai Y.
  • Organizer: EMBO Lecture Courese The 26th.European Cytoskeletal Forum Meeting "Actin-Based Motility"
  • Place of Presentation: Hotel Palma (Stresa,-Lake Maggiore)
  • Year and Date: 2011-10-30
• [Presentation] High-Resolution structural determination of the cofilin-decorated actin filament by single-particle analysis of cryo-electron micrographs (2011)
  • Author(s): Tanaka K., Kimura-Sakiyama C., Dai Shuheng., Maeda Y., Narita A
  • Organizer: EMBO Lecture Courese The 26th.European Cytoskeletal Forum Meeting "Actin-Based Motility"
  • Place of Presentation: Hotel Palma(Stresa,-Lake Maggiore)
  • Year and Date: 2011-10-30
• [Presentation] High-resolution structural determination of the cofilin-decorated actin filament by single-particle analysis of cryo-electron micrographs (2011)
  • Author(s): Tanaka K., Kimura-Sakiyama C., Dai Shuheng., Maeda Y., Narita A.
  • Organizer: 第49回日本生物物理学会
  • Place of Presentation: 兵庫県立大学(姫路市)
  • Year and Date: 2011-09-18
• [Presentation] Visualization of Vibrio Na^+-driven flagellar motor structure embedded in cell membrane by electron microscopy (2011)
  • Author(s): Koike M., Narita A., Honma M., Maeda Y.
  • Organizer: 第49回日本生物物理学会
  • Place of Presentation: 兵庫県立大学(姫路市)
  • Year and Date: 2011-09-18
• [Presentation] A structural analysis of actin cytoskeleton in the cell (2011)
  • Author(s): Narita A., Muller J., Urban E., Vinzenz M., Small J V., Maeda Y.
  • Organizer: 第49回日本生物物理学会
  • Place of Presentation: 兵庫県立大学(姫路市)
  • Year and Date: 2011-09-16
• [Presentation] 細胞骨格のための単粒子解析法"Single particle analysis for cytoskeletal proteins" (2011)
  • Author(s): 成田哲博
  • Organizer: 第11回日本蛋白質科学会年会
  • Place of Presentation: ホテル阪急エキスポパーク(吹田市)(招待講演)
  • Year and Date: 2011-06-07