Mechanism of selective substrate cleqvage and novel cellular functions ofintramembrane protease RseP

• Project/Area Number: 15H04350
• Research Category: Grant-in-Aid for Scientific Research (B)
• Allocation Type: Single-year Grants
• Section: 一般
• Research Field: Functional biochemistry
• Research Institution: Kyoto University
• Principal Investigator: Akiyama Yoshinori 京都大学, ウイルス・再生医科学研究所, 教授 (10192460)
• Co-Investigator(Kenkyū-buntansha): 森 博幸 京都大学, ウイルス・再生医科学研究所, 准教授 (10243271)
• Project Period (FY): 2015-04-01 – 2018-03-31
• Project Status: Completed (Fiscal Year 2017)
• Budget Amount: ¥16,250,000 (Direct Cost: ¥12,500,000、Indirect Cost: ¥3,750,000); Fiscal Year 2017: ¥4,680,000 (Direct Cost: ¥3,600,000、Indirect Cost: ¥1,080,000); Fiscal Year 2016: ¥5,200,000 (Direct Cost: ¥4,000,000、Indirect Cost: ¥1,200,000); Fiscal Year 2015: ¥6,370,000 (Direct Cost: ¥4,900,000、Indirect Cost: ¥1,470,000)
• Keywords: 生体膜 / プロテアーゼ / 膜内切断 / 表層ストレス応答 / 膜タンパク質 / S2P / 細菌 / ストレス応答 / 膜プロテアーゼ / 大腸菌 / 構造生物学

Outline of Final Research Achievements

Intramembrane cleaving proteases (I-CLiPs) are known to act in a variety of important cellular processes in diverse organism. RseP, an Escherichia coli S2P family intramembrane cleaving protease, is involved in regulation of the extracytoplasmic stress response and membrane quality control through specific cleavage of substrates within the lipid bilayer. We have shown that, in addition to the periplasmic PDZ domain that acts as a size-exclusion filter, the cytoplasmic MRE β-loop region and the C1N region play critical roles in specific recognition and substrate cleavage.

Research Products

• [Int'l Joint Research] Monash University(オーストラリア)
• [Journal Article] A new photo-cross-linking approach for analysis of protein dynamics in vivo. (2018)
  • Author(s): Miyazaki, R., Myogo, N., Mori, H. and Akiyama, Y.
  • Journal Title: J. Biol. Chem. Volume: 293 Issue: 2 Pages: 677-686
  • DOI: 10.1074/jbc.m117.817270
  • Peer Reviewed
• [Journal Article] Involvement of a conserved GFG motif region in substrate binding by RseP, an Escherichia coli S2P protease (2017)
  • Author(s): Akiyama, K., Hiuzkuri, Y. and Akiyama, Y.
  • Journal Title: Molecular Microbiology Volume: 印刷中 Issue: 5 Pages: 737-751
  • DOI: 10.1111/mmi.13659
  • Peer Reviewed / Acknowledgement Compliant
• [Journal Article] BepA mediates productive transfer of substrate proteins to the β-barrel assembly machinery (BAM) complex. (2017)
  • Author(s): Daimon, Y., Masui, C., Tanaka, Y., Shiota, T., Suzuki, T., Miyazaki, R., Sakurada, H., Lithgow, T., Dohmae, N., Mori, H., Tsukazaki, T., Narita, S. and Akiyama, Y.
  • Journal Title: Mol. Microbiol. Volume: 106 Issue: 5 Pages: 760-776
  • DOI: 10.1111/mmi.13844
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Biochemical Characterization of Function and Structure of RseP, an Escherichia coli S2P Protease (2017)
  • Author(s): Hizukuri, Y. , Akiyama, K. and Akiyama, Y.
  • Journal Title: Methods in Enzymology Volume: 584 Pages: 1-33
  • DOI: 10.1016/bs.mie.2016.09.044
  • Peer Reviewed / Acknowledgement Compliant
• [Journal Article] A novel SRP recognition sequence in the homeostatic control region od heat shock transcription factor sigma 32 (2016)
  • Author(s): Miyazaki, R., Yura, T., Suzuki, T. Dohmae, N., Mori, H. and Akiyama, Y.
  • Journal Title: Scientfic reports Volume: 6 Issue: 1 Pages: 24147-24147
  • DOI: 10.1038/srep24147
  • Peer Reviewed / Open Access
• [Journal Article] Roles of the membrane-reentrant β-hairpin-like loop of RseP protease in selective substrate cleavage (2015)
  • Author(s): Koichiro Akiyama, Shinya Mizuno, Yohei Hizukuri, Hiroyuki Mori, Terukazu Nogi, Yoshinori Akiyama
  • Journal Title: eLife Volume: 4
  • DOI: 10.7554/elife.08928
  • NAID: 120005712942
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] Nascent chain-monitored remodeling of the Sec machinery for salinity adaptation of marine bacteria (2015)
  • Author(s): Ishii, E., Chiba, S., Hashimoto, N., Kojima, S., Homma, M., Ito, K., Akiyama, Y. and Mori, H.
  • Journal Title: Proc. Natl. Acad. Sci. USA Volume: 112 Issue: 40
  • DOI: 10.1073/pnas.1513001112
  • Peer Reviewed / Acknowledgement Compliant
• [Journal Article] Activation of toxin-antitoxin system toxins suppresses lethality caused by the loss of σE in Escherichia coli. (2015)
  • Author(s): Yasushi Daimon, Shin-ichiro Narita, Yoshinori Akiyama
  • Journal Title: Journal of Bacteriology Volume: 197 Issue: 14 Pages: 2316-2324
  • DOI: 10.1128/jb.00079-15
  • Peer Reviewed
• [Presentation] 大腸菌 S2P ファミリープロテアーゼ RseP の機能未知 PCT 領域の解析 (2017)
  • Author(s): 三宅拓也、檜作洋平、秋山芳展
  • Organizer: 第14回21世紀大腸菌研究会
• [Presentation] Component of the Bacterial Flagellar Type III Secretion System Receives Proteolysis by a Rhomboid Family Protease in the Membrane. (2017)
  • Author(s): 16.Hizukuri, Y., Suzuki, T., Terushima, K., Dohmae, N. and Akiyama, Y.
  • Organizer: International Symposium ‘Harmonized supramolecular motility machinery and its diversity’
  • Int'l Joint Research
• [Presentation] 細菌の表層ストレス応答に関わる膜内切断プロテアーゼ RsePの新規機能調節領域の解析 (2017)
  • Author(s): 29.三宅拓也、檜作洋平、秋山芳展
  • Organizer: 2017年度生命科学系学会合同年次大会
• [Presentation] Regulation of E. coli heat shock response regulator σ32 via SRP protein targeting pathway. (2016)
  • Author(s): Akiyama, Y.
  • Organizer: 23rd East Asia Joint Symposium/15th Cross-Strait Symposium on Biochemical Research/ 13th Symposium of the Frontiers of Biomedical Sciences
  • Place of Presentation: Taipei
  • Year and Date: 2016-10-18
  • Int'l Joint Research / Invited
• [Presentation] 細菌RhomboidプロテアーゼGlpGの生理的切断基質の探索及びべん毛III型分泌装置との関係 (2016)
  • Author(s): 檜作洋平、照島功祐、秋山芳展
  • Organizer: 第89回日本生化学会大会
  • Place of Presentation: senndai
  • Year and Date: 2016-09-25
• [Presentation] S2Pファミリー膜内切断プロテアーゼRsePの膜内挿入ループ領域を介した基質選別 (2015)
  • Author(s): 23.秋山光市郎、水野慎也、檜作洋平、森 博幸、禾 晃和、秋山芳展
  • Organizer: 第38回日本分子生物学会大会・第88回日本生化学会大会合同大会
  • Place of Presentation: 神戸
  • Year and Date: 2015-12-01
• [Presentation] Analysis of the conserved membrane-reentrant loop of RseP, an intramembrane-cleaving protease. (2015)
  • Author(s): 19.Akiyama, K., Mizuno, S., Hizukuri, Y., Mori, H., Nogi, T., Akiyama, Y.
  • Organizer: The 22nd East Asia Joint Symposium on Biomedical Research
  • Place of Presentation: 沖縄
  • Year and Date: 2015-11-11
  • Int'l Joint Research