Molecular mechanism for quality control of bacterial outer membrane by the periplasmic protease BepA

• Project/Area Number: 15K07008
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Research Field: Functional biochemistry
• Research Institution: Morioka College
• Principal Investigator: Narita Shin-ichiro 盛岡大学, 栄養科学部, 教授 (30338751)
• Co-Investigator(Renkei-kenkyūsha): AKIYAMA Yoshinori 京都大学・ウイルス, 再生医科学研究所, 教授 (10192460)
• Research Collaborator: DAIMON Yasushi
• Project Period (FY): 2015-04-01 – 2018-03-31
• Project Status: Completed (Fiscal Year 2017)
• Budget Amount: ¥4,940,000 (Direct Cost: ¥3,800,000、Indirect Cost: ¥1,140,000); Fiscal Year 2017: ¥1,040,000 (Direct Cost: ¥800,000、Indirect Cost: ¥240,000); Fiscal Year 2016: ¥1,690,000 (Direct Cost: ¥1,300,000、Indirect Cost: ¥390,000); Fiscal Year 2015: ¥2,210,000 (Direct Cost: ¥1,700,000、Indirect Cost: ¥510,000)
• Keywords: 細胞内タンパク質分解 / 表層ストレス応答 / 細菌細胞表層 / プロテアーゼ / シャペロン / 外膜タンパク質 / ジスルフィド結合 / タンパク質間相互作用

Outline of Final Research Achievements

BepA is an Escherichia coli periplasmic protease involved in biogenesis and quality control of LptD, a component of the outer membrane lipopolysaccharide translocon. BepA can associate with the BAM complex driving integration of outer membrane proteins into the outer membrane. However, the molecular mechanism of BepA function and its association with the BAM complex remained unknown. In this study, we performed a systematic site-directed in vivo photo-cross-linking analysis to reveal the protein-protein interactions mediated by the BepA TPR domain. We revealed the mode of interaction between the BepA domein and the BAM complex as well as LptD. Mutational analysis indicated that BepA TPR domain is essential and protein-protein interactions mediated by the TPR domain is critical for the function of BepA.

Research Products

• [Int'l Joint Research] Monash University(オーストラリア)
• [Int'l Joint Research] Monash University(オーストラリア)
• [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] Bacterial lipoproteins; biogenesis, sorting and quality control. (2016)
  • Author(s): Shin-ichiro Narita, Hajime Tokuda
  • Journal Title: Biochin. Biophys. Acta Volume: S1388-1981(16) Pages: 30323-30327
  • Peer Reviewed
• [Journal Article] Biogenesis and quality control of the outer membrane of gram-negative bacteria (2015)
  • Author(s): 成田新一郎
  • Journal Title: 生化学 Volume: 87 Issue: 4 Pages: 450-453
  • DOI: 10.14952/SEIKAGAKU.2015.870450
  • NAID: 40020571845
  • ISSN: 0037-1017
  • Year and Date: 2015-08-25
• [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
• [Journal Article] Novel antibacterial targets and compounds revealed by a high throughput cell wall reporter assay (2015)
  • Author(s): Asha S. Nayar, Thomas J. Dougherty, Keith E. Ferguson, Brett A. Granger, Lisa McWilliams, Clare Stacey, Lindsey J. Leach, Shin-ichiro Narita, Hajime Tokuda, Alita A. Miller, Dean G. Brown, and Sarah M. McLeod
  • Journal Title: J. Bacteriol. Volume: 197 Issue: 10 Pages: 1726-1734
  • DOI: 10.1128/jb.02552-14
  • Peer Reviewed
• [Presentation] 大腸菌のペリプラズムプロテアーゼBepAは、TPR domainを介したタンパク質間相互作用により外膜機能を維持する。 (2017)
  • Author(s): 大門康志、舛井千草、宮﨑亮次、櫻田洋人、田中良樹、鈴木健裕、堂前直、森博幸、塚崎智也、成田新一郎、秋山芳展
  • Organizer: 日本農芸化学会2017年度大会
  • Place of Presentation: 京都女子大学
• [Remarks] 研究テーマ
  • URL: https://sites.google.com/site/mununarita/home/yan-jiutema