Molecular mechanisms of AAA proteins revealed by biochemical analyses and high-speed atomic force microscopic observations

• Project/Area Number: 24370056
• Research Category: Grant-in-Aid for Scientific Research (B)
• Allocation Type: Partial Multi-year Fund
• Section: 一般
• Research Field: Functional biochemistry
• Research Institution: Kumamoto University
• Principal Investigator: OGURA Teru 熊本大学, 発生医学研究所, 教授 (00158825)
• Co-Investigator(Renkei-kenkyūsha): OKUNO Takashi 山形大学, 理学部, 准教授 (80411031) ; YAMANAKA Kunitoshi 熊本大学, 発生医学研究所, 准教授 (90212290) ; ESAKI Masatoshi 熊本大学, 発生医学研究所, 助教 (70437911)
• Project Period (FY): 2012-04-01 – 2015-03-31
• Project Status: Completed (Fiscal Year 2014)
• Budget Amount: ¥18,330,000 (Direct Cost: ¥14,100,000、Indirect Cost: ¥4,230,000); Fiscal Year 2014: ¥5,330,000 (Direct Cost: ¥4,100,000、Indirect Cost: ¥1,230,000); Fiscal Year 2013: ¥5,200,000 (Direct Cost: ¥4,000,000、Indirect Cost: ¥1,200,000); Fiscal Year 2012: ¥7,800,000 (Direct Cost: ¥6,000,000、Indirect Cost: ¥1,800,000)
• Keywords: AAAタンパク質 / 分子シャペロン / プロテアソーム / p97/VCP/CDC-48/Cdc48 / Katanin / 高速原子間力顕微鏡 / ミトコンドリア / 微小管 / katanin / p97/CDC-48/Cdc48 / ポリグルタミン凝集体 / 26Sプロテアソーム / 神経変性疾患

Outline of Final Research Achievements

AAA family chaperones/proteases form a hexameric ring structure. ATP-dependent rotation of the AAA chaperone p97 was analyzed by high-speed AFM, and a novel model for the p97 action against substrate has been proposed. It has been found that VCP, a human p97 homolog, binds to amyloid fibrils of TDP-43. UFD-3 is an adapter of CDC-48, a C. elegans p97 homolog, and has been revealed to regulate polyglutamine aggregate formation. Abnormal mitochondrial morphology caused by a mutation of Cdc48, a yeast p97 homolog, was precisely analyzed by advanced microscopic techniques. Dynamics of the 26S proteasome and its complex with a substrate were analyzed by high-speed AFM. Microtubule severing by katanin was analyzed, and a novel model for the mechanism of katanin has been proposed. Functional importance of mitochondrial Bcs1 was revealed.

Research Products

• [Journal Article] Microtubule severing by katanin p60 AAA+ ATPase requires the C-terminal acidic tails of both α- and β-tubulins and basic amino acid residues in the AAA+ ring pore. (2015)
  • Author(s): Johjima, A., Noi, K., Nishikori, S., Ogi, H., Esaki, M., and Ogura, T.
  • Journal Title: J. Biol. Chem. Volume: 印刷中 Issue: 18 Pages: 11762-11770
  • DOI: 10.1074/jbc.m114.614768
  • Peer Reviewed
• [Journal Article] Characterization of C-terminal adaptors, UFD-2 and UFD-3, of CDC-48 on the polyglutamine aggregation in C. elegans. (2015)
  • Author(s): Murayama, Y., Ogura, T., and Yamanaka, K.
  • Journal Title: Biochem. Biophys. Res. Commun. Volume: 459 Issue: 1 Pages: 154-160
  • DOI: 10.1016/j.bbrc.2015.02.088
  • Peer Reviewed
• [Journal Article] Serial block-face scanning electron microscopy for three-dimensional analysis of morphological changes in mitochondria regulated by Cdc48p/p97 ATPase. (2014)
  • Author(s): Miyazaki, N., Esaki, M., Ogura, T., and Murata, K.
  • Journal Title: J. Struct. Biol. Volume: 187 Issue: 2 Pages: 187-193
  • DOI: 10.1016/j.jsb.2014.05.010
  • Peer Reviewed
• [Journal Article] Whole-cell imaging of the budding yeast Saccharomyces cerevisiae by high-voltage scanning transmission electron tomography. (2014)
  • Author(s): Murata, K., Esaki, M., Ogura, T., Arai, S., Yamamoto, Y., and Tanaka, N.
  • Journal Title: Ultramicroscopy Volume: 146 Pages: 39-45
  • DOI: 10.1016/j.ultramic.2014.05.008
  • Peer Reviewed
• [Journal Article] A conserved a helix of Bcs1, a mitochondrial AAA chaperone, is required for the Respiratory Complex III maturation (2014)
  • Author(s): Rie Sawamura, Teru Ogura and Masatoshi Esaki
  • Journal Title: Biochemical and Biophysical Research Communications Volume: 443 Issue: 3 Pages: 997-1002
  • DOI: 10.1016/j.bbrc.2013.12.084
  • Peer Reviewed
• [Journal Article] High-speed atomic force microscopic observation of ATP-dependent rotation of the AAA+ chaperone p97 (2013)
  • Author(s): K. Noi, D. Yamamoto, S. Nishikori, K. Arita-Morioka, T. Ando, T. Ogura
  • Journal Title: Structure Volume: 21 Issue: 11 Pages: 1992-2002
  • DOI: 10.1016/j.str.2013.08.017
  • Peer Reviewed
• [Journal Article] CDC-48/p97 is required for proper meiotic chromosome segregation via controlling AIR-2/Aurora B kinase localization in Caenorhabditis elegans. (2012)
  • Author(s): Sasagawa Y, Higashitani A, Urano T, Ogura T, Yamanaka K.
  • Journal Title: J Struct Biol. Volume: 179 Issue: 2 Pages: 104-111
  • DOI: 10.1016/j.jsb.2012.06.009
  • Peer Reviewed
• [Journal Article] Caenorhabditis elegans fidgetin homolog FIGL-1, a nuclear-localized AAA ATPase, binds to SUMO. (2012)
  • Author(s): Onitake, A., Yamanaka, K., Esaki, M., and Ogura, T.
  • Journal Title: J. Struct. Biol. Volume: (in press) Issue: 2 Pages: 143-151
  • DOI: 10.1016/j.jsb.2012.04.022
  • Peer Reviewed
• [Journal Article] The C-terminal -helix of SPAS-1, a Caenorhabditis elegans spastin homologue, is crucial for microtubule severing. (2012)
  • Author(s): Onitake, A., Matsushita-Ishiodori, Y., Johjima, A., Esaki, M., Ogura, T., and Yamanaka, K.
  • Journal Title: J. Struct. Biol. Volume: (in press) Issue: 2 Pages: 138-142
  • DOI: 10.1016/j.jsb.2012.04.010
  • Peer Reviewed
• [Journal Article] Cdc48p/p97-mediated regulation of mitochondrial morphology is Vms1p independent. (2012)
  • Author(s): Esaki, M., and Ogura, T.
  • Journal Title: J. Struct. Biol. Volume: (in press) Issue: 2 Pages: 112-120
  • DOI: 10.1016/j.jsb.2012.04.017
  • Peer Reviewed
• [Presentation] ホウレンソウ由来天然FtsHの精製と解析 (2014)
  • Author(s): 鈴木 香於里、大枝 弘明、加藤 裕介、有田 (森岡）健一、野井 健太郎、原 怜、山本 亘、小椋 光、久堀 徹、天野 豊己
  • Organizer: 日本生体エネルギー研究会 第40回討論会
  • Place of Presentation: 松山市 愛媛大学 南加記念ホール
  • Year and Date: 2014-12-12 – 2014-12-13
• [Presentation] ヒト疾患関連AAAタンパク質BCS1Lの酵母ホモログBcs1pの機能解析 (2014)
  • Author(s): 澤村 理英、江崎 雅俊、山中 邦俊、小椋 光
  • Organizer: 第37回日本分子生物学会年会
  • Place of Presentation: 横浜市 パシフィコ横浜
  • Year and Date: 2014-11-25 – 2014-11-27
• [Presentation] ポリグルタミン凝集体形成における線虫CDC-48 C末端アダプターの役割 (2014)
  • Author(s): 村山 佑樹、小椋 光、山中 邦俊
  • Organizer: 第37回日本分子生物学会年会
  • Place of Presentation: 横浜市 パシフィコ横浜
  • Year and Date: 2014-11-25 – 2014-11-27
• [Presentation] AAAタンパク質kataninによる微小管切断におけるAAAドメインのpore領域の残基とtubulin C末端配列の重要性 (2014)
  • Author(s): 城島 愛、野井 健太郎、錦織 伸吾、荻 博次、江崎 雅俊、小椋 光
  • Organizer: 第37回日本分子生物学会年会
  • Place of Presentation: 横浜市 パシフィコ横浜
  • Year and Date: 2014-11-25 – 2014-11-27
• [Presentation] Cdc48/p97はFzo1の分解促進によってミトコンドリア融合反応を制御する (2014)
  • Author(s): 江崎 雅俊、宮崎 直幸、村田 和義、小椋 光
  • Organizer: 第37回日本分子生物学会年会
  • Place of Presentation: 横浜市 パシフィコ横浜
  • Year and Date: 2014-11-25 – 2014-11-27
• [Presentation] 高速原子間力顕微鏡によるAAAシャペロンp97の主要ATPase(D2)リングの構造変化の直接観察 (2014)
  • Author(s): 山本 大輔、野井 健太郎、有田 (森岡) 健一、小椋 光
  • Organizer: 第52回 日本生物物理学会年会
  • Place of Presentation: 札幌市 札幌コンベンションセンター
  • Year and Date: 2014-09-25 – 2014-09-27
• [Presentation] Visualization of dynamics of the 26S proteasome and proteasome-substrate complexes by high-speed AFM (2014)
  • Author(s): Okuno, T., Noi, K., Okawa, A., Tsuchiya, H., Saeki, Y., Takahashi, K., Inobe, T., Yamanaka, K., and Ogura, T.
  • Organizer: Key Forum: From Stem Cells to Organs
  • Place of Presentation: 熊本市 熊本市医師会館
  • Year and Date: 2014-09-04 – 2014-09-05
• [Presentation] 高速原子間力顕微鏡によるAAAシャペロンp97の主要ATPase(D2)リングの直接観察 (2014)
  • Author(s): 山本 大輔、野井 健太郎、有田 (森岡) 健一、小椋 光
  • Organizer: 第14回 日本蛋白質科学会年会
  • Place of Presentation: 横浜市 ワークピア横浜/横浜産貿ホールマリネリア
  • Year and Date: 2014-06-25 – 2014-06-27
• [Presentation] Adaptors of p97 and disease-associated p97 mutations. (2014)
  • Author(s): Yamanaka, K., Murayama, Y., and Ogura, T.
  • Organizer: 231st IMEG seminars & Minisymposium
  • Place of Presentation: 熊本市 熊本大学発生医学研究所
• [Presentation] Yeast Cdc48-20S proteasome complex in vitro and in vivo. (2014)
  • Author(s): Esaki, M., Noi, K., and Ogura, T.
  • Organizer: 231st IMEG seminars & Minisymposium
  • Place of Presentation: 熊本市 熊本大学発生医学研究所
• [Presentation] High-speed AFM imaging of the 26S proteasome dynamics. (2014)
  • Author(s): Okuno, T., Noi, K., Okawa, A., Tsuchiya, H., Saeki, Y., Takahashi, K., Inobe, T., Yamanaka, K., and Ogura, T.
  • Organizer: 231st IMEG seminars & Minisymposium
  • Place of Presentation: 熊本市 熊本大学発生医学研究所
• [Presentation] Observation of AAA chaperone p97 by high-speed AFM. (2014)
  • Author(s): Yamamoto, D., Noi, K., Arita-Morioka, K, Ando, T., and Ogura, T.
  • Organizer: 231st IMEG seminars & Minisymposium
  • Place of Presentation: 熊本市 熊本大学発生医学研究所
• [Presentation] Clamp loading by Gamma complex. (2014)
  • Author(s): Karata, K., and Ogura, T.
  • Organizer: 231st IMEG seminars & Minisymposium
  • Place of Presentation: 熊本市 熊本大学発生医学研究所
• [Presentation] Biochemical properties of FtsH protease from plants. (2014)
  • Author(s): Ueda, E., Noi, K., Arita-Morioka, K., Ogura, T., Sakamoto, W., Hisabori, T., and Amano, T.
  • Organizer: 231st IMEG seminars & Minisymposium
  • Place of Presentation: 熊本市 熊本大学発生医学研究所
• [Presentation] High-speed AFM observation of ATP-dependent rotation of the AAA chaperone p97 and its interaction with TDP-43 amyloid fibrils. (2014)
  • Author(s): Noi, K., Yamamoto, D., Arita-Morioka, K., Ando, T., and Ogura, T.
  • Organizer: 231st IMEG seminars & Minisymposium
  • Place of Presentation: 熊本市 熊本大学発生医学研究所
• [Presentation] Dynamics of p97 and the 26S proteasome in action revealed by high-speed atomic force microscopy. (2013)
  • Author(s): Ogura, T., Noi, K., Okuno, T., Arita-Morioka, K., Yamamoto, D., Okawa, A., Tsuchiya, H., Saeki, Y., Ando, T., Esaki, M., and Yamanaka, K.
  • Organizer: EMBO Workshop on AAA+ Proteins
  • Place of Presentation: Commundo Hotel, Neuss, Germany.
  • Invited
• [Presentation] Functional analysis of disease-associated mutant CDC-48/p97/VCP in C. elegans. (2013)
  • Author(s): Yamanaka, K., Murayama, Y., and Ogura, T.
  • Organizer: EMBO Workshop on AAA+ Proteins
  • Place of Presentation: Commundo Hotel, Neuss, Germany.
• [Presentation] Non-consevation of the aromatic residue in the D1 pore loop of yeast Cdc48 is essential for cell viability. (2013)
  • Author(s): Esaki, M., and Ogura, T.
  • Organizer: EMBO Workshop on AAA+ Proteins
  • Place of Presentation: Commundo Hotel, Neuss, Germany.
• [Presentation] Biochemical analysis of molecular mechanisms of the interaction between VCP/p97 and amyloid fibrils of TDP-43. (2013)
  • Author(s): Noi, K., Arita-Morioka, K., Ogi, H., Hirao, M., and Ogura, T.
  • Organizer: EMBO Workshop on AAA+ Proteins
  • Place of Presentation: Commundo Hotel, Neuss, Germany.
• [Presentation] The N-terminal region of Bcs1p is important for the respiratory chain complex III assembly. (2013)
  • Author(s): Sawamura, R., Esaki, M., and Ogura, T.
  • Organizer: EMBO Workshop on AAA+ Proteins
  • Place of Presentation: Commundo Hotel, Neuss, Germany.
• [Presentation] AAA chaperones, focusing on the molecular mecanism of p97. (2013)
  • Author(s): Ogura, T., Noi, K., Arita-Morioka, K., Yamamoto, D., Ando, T., Nishikori, S., Esaki, M., and Yamanaka, K.
  • Organizer: EMBO Conference The biology of molecular chaperones
  • Place of Presentation: Margherita de Pula, Italy
  • Invited
• [Presentation] 植物由来FtsHプロテアーゼの分子機構. (2013)
  • Author(s): 斉藤 勝広, 植田 依里, 柿本 衿菜, 鈴木 香於里, 原 怜, 久堀 徹, 加藤 裕介, 坂本 亘, 野井 健太郎, 有田 健一, 小椋 光, 天野 豊己
  • Organizer: 日本生体エネルギー研究会 第39回討論会
  • Place of Presentation: 静岡市, 静岡県コンベンションアーツセンター
• [Presentation] 線虫を用いた疾患型変異CDC-48/p97/VCPの機能解析 (2013)
  • Author(s): 山中 邦俊, 村山 佑樹, 小椋 光
  • Organizer: 第36回日本分子生物学会年会
  • Place of Presentation: 神戸市、神戸国際展示場
• [Presentation] In vivo においてCdc48は20Sプロテアソーム複合体と相互作用する. (2013)
  • Author(s): 江崎 雅俊, 野井 健太郎, 佐伯 泰, 小椋 光
  • Organizer: 第36回日本分子生物学会年会
  • Place of Presentation: 神戸市、神戸国際展示場
• [Presentation] 呼吸鎖複合体III形成におけるAAA分子シャペロンBcs1pのN末端の機能解析. (2013)
  • Author(s): 澤村 理英, 江崎 雅俊, 小椋 光
  • Organizer: 第36回日本分子生物学会年会
  • Place of Presentation: 神戸市、神戸国際展示場
• [Presentation] 高速AFMによる26Sプロテアソームのダイナミクス解析. (2013)
  • Author(s): 奥野 貴士, 野井 健太郎, 大川 阿紀子, 土屋 光, 佐伯 泰, 高橋 一暢, 伊野部 智由, 山中 邦俊, 小椋 光
  • Organizer: 第36回日本分子生物学会年会
  • Place of Presentation: 神戸市、神戸国際展示場
• [Presentation] 分子シャペロンVCP/p97とTDP-43アミロイド線維の相互作用の分子機構の解析. (2013)
  • Author(s): 野井 健太郎, 有田ー森岡 健一, 小椋 光
  • Organizer: 第13回日本蛋白質科学会年会
  • Place of Presentation: 鳥取市、とりぎん文化会館
• [Presentation] High-speed atomic force microscopy of ATP-dependent rotational movementsof p97/VCP and its interaction with amyloid fibrils of TDP-43. (2012)
  • Author(s): Noi, K., Yamamoto, D., Arita-Morioka, K., Nishikori, S., Ando, T., and Ogura, T.
  • Organizer: 3rd Kanazawa Bio-AFM Workshop
  • Place of Presentation: 金沢・KKRホテル金沢
• [Presentation] 分子シャペロンVCP/p97とTDP-43アミロイド線維の相互作用の分子機構の解析. (2012)
  • Author(s): 野井 健太郎,有田・森岡 健一,小椋 光
  • Organizer: 第85回日本生化学会大会
  • Place of Presentation: 福岡・福岡国際会議場,マリンメッセ福岡
• [Book] Subcell. Biochem (2013)
  • Author(s): Okuno, T., and Ogura, T.
  • Total Pages: 17
  • Publisher: Springer
• [Book] Handbook of Proteolytic Enzymes, 3rd edition (2013)
  • Author(s): Ogura, T., Okuno, T., Suno, R., and Akiyama, Y.
  • Total Pages: 8
  • Publisher: Elsevier Ltd.
• [Book] FtsH protease-mediated regulation of various cellular functions. Subcell.Biochem, 66, Regulated proteolysis in microorganisms. D. A. Dougan (ed.) (2013)
  • Author(s): Okuno, T., and Ogura, T.
  • Total Pages: 17
  • Publisher: Springer
• [Book] FtsH protease. In Handbook of Proteolytic Enzymes, 3rd edition, N. Rawlings andG. Salvesen (ed.) (2013)
  • Author(s): Ogura, T., Okuno, T., Suno, R., and Akiyama, Y.
  • Total Pages: 8
  • Publisher: Elsevier Ltd.
• [Remarks] 発生医学研究所分子細胞制御分野のホームページ
  • URL: http://mukb.medic.kumamoto-u.ac.jp/fukusei/index.html
• [Remarks] 発生医学研究所分子細胞制御分野AAAのホームページ
  • URL: http://mukb.medic.kumamoto-u.ac.jp/AAA/aaainfo.html
• [Remarks] 発生医学研究所のホームページ
  • URL: http://www.imeg.kumamoto-u.ac.jp/index.html
• [Remarks] 熊本大学発生医学研究所分子細胞制御分野のホームページ
  • URL: http://mukb.medic.kumamoto-u.ac.jp/fukusei/index.html
• [Remarks] 熊本大学発生医学研究所分子細胞制御分野 AAAホームページ
  • URL: http://mukb.medic.kumamoto-u.ac.jp/AAA/aaainfo.html
• [Remarks] 熊本大学発生医学研究所のホームページ
  • URL: http://www.imeg.kumamoto-u.ac.jp/index.html
• [Remarks] 熊本大学発生医学研究所分子細胞制御分野のホームページ
  • URL: http://mukb.medic.kumamoto-u.ac.jp/fukusei/index.html
• [Remarks] 熊本大学発生医学研究所分子細胞制御分野 AAAホームページ
  • URL: http://mukb.medic.kumamoto-u.ac.jp/AAA/aaainfo.html
• [Remarks] 熊本大学発生医学研究所のホームページ
  • URL: http://www.imeg.kumamoto-u.ac.jp/index.html