Elucidation of the mode of action of Atg12-Atg5 conjugate in autophagy

• Project/Area Number: 24770092
• Research Category: Grant-in-Aid for Young Scientists (B)
• Allocation Type: Multi-year Fund
• Research Field: Structural biochemistry
• Research Institution: Tokyo Institute of Technology
• Principal Investigator: NAKATOGAWA Machiko 東京工業大学, フロンティア研究機構, 先進研究員 (90402461)
• Project Period (FY): 2012-04-01 – 2014-03-31
• Project Status: Completed (Fiscal Year 2013)
• Budget Amount: ¥4,680,000 (Direct Cost: ¥3,600,000、Indirect Cost: ¥1,080,000); Fiscal Year 2013: ¥2,470,000 (Direct Cost: ¥1,900,000、Indirect Cost: ¥570,000); Fiscal Year 2012: ¥2,210,000 (Direct Cost: ¥1,700,000、Indirect Cost: ¥510,000)
• Keywords: ユビキチン様タンパク質 / E3酵素 / オートファジー / 構造変化 / E2酵素 / 結晶構造解析

Research Abstract

Autophagy is a bulk degradation system, which involves the formation of a double-membrane vesicle called autophagosome. Autophagosome formation requires two ubiquitin-like proteins, Atg8 and Atg12. They are conjugated to the phosphatidylethanolamine (PE) and Atg5, respectively, via a series of enzymatic reactions with E1 and E2 enzyme. In this study, we elucidated the mode of action of Atg12-Atg5 as an E3 enzyme in the Atg8-PE conjugation reaction. We established a biochemical assay based on the structural information to determine the configuration of the catalytic center of Atg3, which is an E2 enzyme in Atg8-PE conjugation reaction. This approach revealed that Atg12-Atg5 conjugate induces a conformational change in the catalytic center of Atg3 to enhance its E2 activity. Moreover, mutational analyses indicated how the activity of Atg3 is suppressed in the absence of Atg12-Atg5 conjugate. We are attempting to analyze the crystal structure of Atg12-Atg5 conjugate/Atg3 complex.

Research Products

• [Journal Article] Atg12-Atg5 conjugate enhance E2 activity of Atg3 by rearranging its catalytic site (2013)
  • Author(s): Sakoh-Nakatogawa M., Matoba K., Asai E., Kirisako H., Ishii J., Noda N. N., Inagaki F., Nakatogawa H., Ohsumi Y.
  • Journal Title: Nat. Struct. Mol. Biol. Volume: 20 Pages: 433-439
  • Peer Reviewed
• [Journal Article] Atg12-Atg5 conjugate enhances E2 activity of Atg3 by rearranging its catalytic site (2013)
  • Author(s): Sakoh-Nakatogawa, M., Matoba, K., Asai, E., Kirisako, H., Ishii, J., Noda, N. N., Inagaki, F., Nakatogawa, H. and Ohsumi, Y
  • Journal Title: Nature Structural & Molecular Biology Volume: Vol.20 Issue: 4 Pages: 433-439
  • DOI: 10.1038/nsmb.2527
  • Peer Reviewed
• [Journal Article] The autophagy-related protein kinase Atg1 interacts with the ubiquitin-like protein Atg8 via the Atg8 family interacting motif to facilitate autophagosome formation (2012)
  • Author(s): Nakatogawa H., Ohbayashi S., Sakoh-Nakatogawa M., Kakuta S., Suzuki S. W., Kirisako H., Kondo-Kakuta C., Noda N. N., Yamamoto H., Ohsumi Y.
  • Journal Title: J. Biol. Chem. Volume: 287 Pages: 28503-28507
  • Peer Reviewed
• [Presentation] オートファゴソーム形成を駆動するユビキチン様タンパク質脂質化反応のメカニズム (2014)
  • Author(s): 中戸川万智子、的場一晃、野田展生、稲垣冬彦、中戸川仁、大隅良典
  • Organizer: 日本蛋白質科学会年会
  • Place of Presentation: ワークピア横浜／横浜産貿ホール（神奈川）
  • Invited
• [Presentation] Regulation of lipidation of the ubiquitin-like protein Atg8 that drives autophagosome formation (2014)
  • Author(s): Machiko Sakoh-Nakatogawa, Kazuaki Matoba, Nobuo Noda, Fuyuhiko Inagaki, Hitoshi Nakatogawa, Yoshinori Ohsumi
  • Organizer: The 4th Asia Pacific Protein Association (APPA) Conference
  • Place of Presentation: 済州島（韓国）
  • Invited
• [Presentation] A novel type of E3 enzyme : the autophagy-related ubiquitin-like protein conjugate Atg12-Atg5 induces a conformational change in the E2 enzyme Atg3 to enhance its conjugase activity (2013)
  • Author(s): Sakoh-Nakatogawa M., Matoba K., Noda N. N., Nakatogawa H., Ohsumi Y.
  • Organizer: The 35^<th> Naito Conference
  • Place of Presentation: Sapporo, Japan
  • Year and Date: 2013-07-11
• [Presentation] A novel type of E3 enzyme: the autophagy-related ubiquitin-like protein conjugate Atg12-Atg5 induces a conformational change in the E2 enzyme Atg3 to enhance its conjugase activity (2013)
  • Author(s): Machiko Sakoh-Nakatogawa, Kazuaki Matoba, Nobuo Noda, Hitoshi Nakatogawa, Yoshinori Ohsumi
  • Organizer: The 35th Naito Conference
  • Place of Presentation: シャトレーゼガトーキングダム（北海道）
• [Presentation] オートファジーに関わる2つのユビキチン様タンパク質結合系のクロストーク (2012)
  • Author(s): 中戸川万智子、中戸川仁、大隅良典
  • Organizer: 日本分子生物学会年会
  • Place of Presentation: 福岡
  • Year and Date: 2012-12-11
• [Presentation] オートファジーに必須のE3酵素Atg12-Atg5によるE2酵素Atg3の活性化機構 (2012)
  • Author(s): 中戸川万智子、中戸川仁、大隅良典
  • Organizer: 日本蛋白質科学会若手奨励賞シンポジウム
  • Place of Presentation: 名古屋
  • Year and Date: 2012-06-21
• [Presentation] The E3 enzyme Atg12-Atg5 rearranges the catalytic center of Atg3 to enhance its E2 activity (2012)
  • Author(s): Sakoh-Nakatogawa M., Nakatogawa H., Ohsumi Y.
  • Organizer: The 6^<th> International Symposium on Autophagy 2012
  • Place of Presentation: Okinawa, Japan
• [Presentation] オートファジーに必須のE3酵素Atg12‐Atg5によるE2酵素Atg3の活性化機構 (2012)
  • Author(s): 中戸川万智子・中戸川 仁・大隅良典
  • Organizer: 日本蛋白質科学会年会
  • Place of Presentation: 名古屋国際会議場（愛知県）
• [Presentation] The E3 enzyme Atg12-Atg5 rearranges the catalytic center of Atg3 to enhace its E2 activity (2012)
  • Author(s): Machiko Sakoh-Nakatogawa, Hitoshi Nakatogawa, Yoshinori Ohsumi
  • Organizer: The 6th international symposium on autophagy 2012
  • Place of Presentation: 万国津梁館（沖縄県）
• [Presentation] オートファジーに関わる２つのユビキチン様タンパク質結合系のクロストーク (2012)
  • Author(s): 中戸川万智子
  • Organizer: 第３５回日本分子生物学会年会
  • Place of Presentation: 福岡国際会議場（福岡県）
  • Invited
• [Remarks] 東工大プレスリリース「オートファジーに必須な因子の作用機構を分子レベルで解明-神経変性疾患や肝疾患の創薬につながる成果-」
  • URL: http://www.titech.ac.jp/pressrelease/
• [Remarks] ライフサイエンス新着論文レビュー(First Author's)「ユビキチン様タンパク質Atg12-Atg5結合体はAtg3の活性中心の再編成をひき起こしその酵素活性を上昇させる」
  • URL: http://first.lifesciencedb.jp/archives/6807
• [Remarks] ユビキチン様タンパク質Atg12-Atg5結合体
  • URL: http://first.lifesciencedb.jp/archives/6807
• [Remarks] オートファジーに必須な因子の作用機構を分子レベルで解明
  • URL: http://www.titech.ac.jp/file/pr20130422_osumi.pdf