Redox regulation of proteolysis in the Enterobacteriacea cells

• Project/Area Number: 15K07019
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Research Field: Functional biochemistry
• Research Institution: Institute of Physical and Chemical Research
• Principal Investigator: Nishii Wataru 国立研究開発法人理化学研究所, 横山構造生物学研究室, 研究員 (30287461)
• Project Period (FY): 2015-04-01 – 2018-03-31
• Project Status: Completed (Fiscal Year 2017)
• Budget Amount: ¥5,200,000 (Direct Cost: ¥4,000,000、Indirect Cost: ¥1,200,000); Fiscal Year 2017: ¥1,560,000 (Direct Cost: ¥1,200,000、Indirect Cost: ¥360,000); Fiscal Year 2016: ¥1,560,000 (Direct Cost: ¥1,200,000、Indirect Cost: ¥360,000); Fiscal Year 2015: ¥2,080,000 (Direct Cost: ¥1,600,000、Indirect Cost: ¥480,000)
• Keywords: タンパク質分解 / 細胞内プロテオリシス / AAA+プロテアーゼ / レドックス / 腸内細菌科 / プロテオリシス / X線結晶解析 / X線小角散乱解析 / 細胞内タンパク質分解 / X線結晶構造 / 腸内細菌 / 結晶構造 / レドックス制御 / アロステリックジスルフィド結合

Outline of Final Research Achievements

Cellular proteolysis dominates function, turnover, and quality of cellular proteins. Lon protease involves such proteolysis in bacterial cytosols and eukaryotic organella, and carries out a series of protein degradation processes within a single molecule. In this study, I investigated Lon protease by structural and functional analyses, including X-ray crystallography. The results showed that function of Lon protease is regulated through its dynamic structural changes in a multi-layered and stepwise manner. The results also shed light on the general mechanisms for cellular proteolysis, and provide opportunities to develop therapy, including that for infectious diseases.

Research Products

• [Journal Article] SARS-CoV 3CL protease cleaves its C-terminal autoprocessing site by novel subsite cooperativity (2016)
  • Author(s): Muramatsu, T., Takemoto, C., Kim, Y.-T., Wang, H., Nishii, W., Terada, T. , Shirouzu, M., Yokoyama, S.
  • Journal Title: Proc. Natl Acad. Sci. USA Volume: 113 Pages: 12997-13002
  • Peer Reviewed
• [Presentation] ダイナミックな構造変化を介したAAA+プロテアーゼLonの制御メカニズム (2017)
  • Author(s): 西井亘、藤井佳史、松村義隆、仙石徹、小島正樹、横山茂之
  • Organizer: 生物学会年会・第90回日本生化学会大会 合同大会（ConBio2017）
• [Presentation] AAA+プロテアーゼLonの分子メカニズム (2016)
  • Author(s): 西井亘，新野陸子，寺田貴帆，白水美香子，村松知成，横山茂之

  • Organizer: 第39回日本分子生物学会年会・公募シンポジウム
  • Place of Presentation: パシフィコ横浜（横浜市）
  • Year and Date: 2016-12-01
• [Presentation] Redox regulation of Lon protease activity by a disulfide bond in facultative anaerobes (2016)
  • Author(s): Nishii, W., Seki, E., Yanagisawa, T., Sehngoku, T., Terada, T., Murmatsu, T., Niino-Kukimoto, M, Shirouzu, M., Kojima, M., Kihara, H. & Yokoyama, S.
  • Organizer: FASEB Science Research Conference
  • Place of Presentation: コロラド（USA）
  • Year and Date: 2016-08-17
  • Int'l Joint Research / Invited
• [Presentation] Production of hepatitis C virus NS2-3 protease by the Escherichia coli cell-free protein synthesis method and control of its activity (2015)
  • Author(s): 西井亘，松本武久，村松知成，横山茂之
  • Organizer: 第38回日本分子生物学会年会・第88回日本生化学会大会合同大会
  • Place of Presentation: 神戸ポートアイランド（兵庫県神戸市）
  • Year and Date: 2015-12-01
• [Presentation] A redox switch shapes the exit pore of Enterobacteriaceal Lon protease to facultatively regulate proteolysis (2015)
  • Author(s): Nishii, W., Niino-Kukimoto, M, Terada, T., Shirouzu, M. Muramatsu, T., & Yokoyama, S.

  • Organizer: 9th General Meeting of the International Proteolysis Society
  • Place of Presentation: ペナン（マレーシア）
  • Year and Date: 2015-10-06
  • Int'l Joint Research
• [Presentation] アロステリックジスルフィド結合による細胞内プロテオリシスのレドックス制御 (2015)
  • Author(s): 西井亘，横山茂之
  • Organizer: 第15回日本蛋白質科学会年会公募型シンポジウム
  • Place of Presentation: あわぎんホール（徳島）
  • Year and Date: 2015-06-26
• [Remarks] 理化学研究所プレスリリース
  • URL: http://www.riken.jp/pr/press/2014/20141111_2/