A novel regulation mechanism of cellular functions by intramembrane proteolysis

• Project/Area Number: 24370054
• Research Category: Grant-in-Aid for Scientific Research (B)
• Allocation Type: Partial Multi-year Fund
• Section: 一般
• Research Field: Functional biochemistry
• Research Institution: Kyoto University
• Principal Investigator: AKIYAMA Yoshinori 京都大学, ウイルス研究所, 教授 (10192460)
• Co-Investigator(Kenkyū-buntansha): MORI Hiroyuki 京都大学, ウイルス研究所, 准教授 (10243271)
• 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: ¥5,460,000 (Direct Cost: ¥4,200,000、Indirect Cost: ¥1,260,000); Fiscal Year 2013: ¥5,330,000 (Direct Cost: ¥4,100,000、Indirect Cost: ¥1,230,000); Fiscal Year 2012: ¥7,540,000 (Direct Cost: ¥5,800,000、Indirect Cost: ¥1,740,000)
• Keywords: 大腸菌 / 膜プロテアーゼ / 膜内タンパク質切断 / 表層ストレス応答 / S2Pプロテアーゼ / 基質探索 / PDZドメイン

Outline of Final Research Achievements

The Escherichia coli σE extracytoplasmic stress response monitors and responds to folding stress in the cell envelope. A protease cascade directed at RseA, a membrane-spanning anti-σ that inhibits σE activity, controls this critical signal-transduction system. Stress cues activate DegS to cleave RseA; a second cleavage by RseP releases RseA from the membrane, enabling its rapid degradation.
We also analyzed the three-dimensional structure of the two tandemly arranged PDZ domains (PDZ tandem) present in the periplasmic region of RseP. Our results suggest that the PDZ tandem serves as a size-exclusion filter to accommodate the truncated form of RseA into the active center.

Research Products

• [Journal Article] A structure-based model of substrate discrimination by a non-canonical PDZ tandem in the intramembrane-cleaving protease RseP. (2014)
  • Author(s): 檜作 洋平, 小田 隆, 田畑 早苗, 川上-田村 恵子, 大井 里香, 佐藤 衛, 高木 淳一, 秋山 芳展, 禾 晃和
  • Journal Title: Structure Volume: 22 Issue: 2 Pages: 326-336
  • DOI: 10.1016/j.str.2013.12.003
  • Peer Reviewed
• [Journal Article] Expression and purification of integral membrane metallopeptidase HtpX (2014)
  • Author(s): Arolas, J. L., García-Castellanos, R., Goulas, T., Akiyama, Y. and Gomis-Rüth, F. X.
  • Journal Title: Protein Expr. Purif. Volume: 99C Pages: 113-118
  • DOI: 10.1016/j.pep.2014.04.008
  • Peer Reviewed
• [Journal Article] PDZ domains of RseP are not essential for sequential cleavage of RseA or stress-induced σ^E activation in vivo (2012)
  • Author(s): 檜作 洋平, 秋山 芳展
  • Journal Title: Molecular Microbiology Volume: 86 Issue: 5 Pages: 1232-1245
  • DOI: 10.1111/mmi.12053
  • Peer Reviewed
• [Presentation] 細菌表層ストレス応答制御に関わる膜内切断プロテアーゼRsePのタンデムPDZドメインによる切断基質選別機構の解析 (2014)
  • Author(s): 檜作洋平、小田 隆、田畑早苗、川上-田村恵子、大井里香、佐藤 衛、高木淳一、禾 晃和、秋山芳展
  • Organizer: 第87回日本生化学会大会
  • Place of Presentation: 京都
  • Year and Date: 2014-10-15 – 2014-10-18
• [Presentation] Substrate discrimination by size-exclusion in the intramembrane protease RseP (2014)
  • Author(s): 13.Hizukuri, Y., Oda, T., Tabata, S., Tamura-Kawakami, K., Oi, R., Sato, M., Takagi, J., Akiyama, Y., and Nogi, T.
  • Organizer: IUCr 2014 - 23rd Congress and General Assembly
  • Place of Presentation: Montreal, Canada
  • Year and Date: 2014-08-05 – 2014-08-12
• [Presentation] 大腸菌膜内切断プロテアーゼRsePの基質認識におけるPDZドメインの役割. (2013)
  • Author(s): 檜作洋平、禾 晃和、小田隆、田畑早苗、川上-田村恵子、佐藤衛、高木淳一、秋山芳展
  • Organizer: 第８６回日本生化学会大会
  • Place of Presentation: 横浜
  • Invited
• [Presentation] 大腸菌RIPプロテアーゼRsePのタンデムPDZドメインによる新たな機能制御メカニズム. (2012)
  • Author(s): 檜作洋平, 他
  • Organizer: 第85回日本生化学会大会
  • Place of Presentation: 福岡
  • Year and Date: 2012-12-14
• [Remarks] 京都大学ウイルス研究所・がん遺伝子分野
  • URL: http://www.virus.kyoto-u.ac.jp/Lab/akiyama/index.html
• [Remarks] がん遺伝子分野
  • URL: http://www.virus.kyoto-u.ac.jp/Lab/akiyama/index.html
• [Remarks]
  • URL: http://www.virus.kyoto-u.ac.jp/Lab/akiyama/index.html