Analysis of bacterial effector proteins that negatively regulate the host ubiquitin system

• Project/Area Number: 16H05189
• Research Category: Grant-in-Aid for Scientific Research (B)
• Allocation Type: Single-year Grants
• Section: 一般
• Research Field: Bacteriology (including mycology)
• Research Institution: Gifu University (2017-2018); Osaka University (2016)
• Principal Investigator: Kubori Tomoko 岐阜大学, 大学院医学系研究科, 准教授 (20397657)
• Project Period (FY): 2016-04-01 – 2019-03-31
• Project Status: Completed (Fiscal Year 2018)
• Budget Amount: ¥17,550,000 (Direct Cost: ¥13,500,000、Indirect Cost: ¥4,050,000); Fiscal Year 2018: ¥5,980,000 (Direct Cost: ¥4,600,000、Indirect Cost: ¥1,380,000); Fiscal Year 2017: ¥5,460,000 (Direct Cost: ¥4,200,000、Indirect Cost: ¥1,260,000); Fiscal Year 2016: ¥6,110,000 (Direct Cost: ¥4,700,000、Indirect Cost: ¥1,410,000)
• Keywords: ユビキチン / 脱ユビキチン化酵素 / レジオネラ / エフェクター / 細菌感染

Outline of Final Research Achievements

Legionella is a bacterial pathogen that transports an extraordinary amount of effector proteins into host eukaryotic cells to manipulate various cellular systems. We have identified Legionella deubiquitinases (DUBs) as negative regulators of the host ubiquitin system. LotA was found to have dual catalytic cysteine residues and to remove ubiquitin from bacterial vacuoles in the infected cells. The DUB activity of LotA contributes intracellular bacterial growth in conjunction with other bacterial ubiquitin ligases. We also identified another Legionella DUB and its cellular target. These results demonstrate that Legionella possesses multiple approaches to manipulate host ubiquitin systems for bacterial benefit.

Academic Significance and Societal Importance of the Research Achievements

病原細菌が感染を確立する上で、宿主真核細胞の持つ普遍的なタンパク質修飾系であるユビキチンシステムを操作・撹乱することは重要な戦略として理解されている。しかしながら、ユビキチン化の負の制御機構はほとんどわかっていなかった。多様なエフェクターレパートリーを持つレジオネラをモデルシステムとして複数の脱ユビキチン化酵素を同定し、その感染における役割を分子レベルで明らかにした本研究課題は、真核生物との共存の中で確立された病原細菌の巧妙な戦略の理解に飛躍的前進を与えるものである。さらに、細菌が正負両面からユビキチン系を操作する仕組みの理解は、従来とは異なる視点からの感染防御への手がかりを与えるものである。

Research Products

• [Journal Article] Emerging insights into bacterial deubiquitinases (2019)
  • Author(s): Tomoko kubori, Tomoe Kitao, Hiroki Ando
  • Journal Title: Current Opinion in Microbiology Volume: 47 Pages: 14-19
  • DOI: 10.1016/j.mib.2018.10.001
  • Peer Reviewed
• [Journal Article] Isolation of the Dot/Icm Type IV Secretion System Core Complex from Legionella pneumophila. (2019)
  • Author(s): Kubori T, Nagai H.
  • Journal Title: Methods Mol Biol. Volume: 1921 Pages: 241-247
  • DOI: 10.1007/978-1-4939-9048-1_15
  • ISBN: 9781493990474, 9781493990481
  • Peer Reviewed
• [Journal Article] LotA, a Legionella deubiquitinase, has dual catalytic activity and contributes to intracellular growth. (2018)
  • Author(s): Kubori T, Kitao T, Ando H and Nagai H.
  • Journal Title: Cell Microbiol. Volume: Mar 15 Issue: 7 Pages: e12840-e12840
  • DOI: 10.1111/cmi.12840
  • Peer Reviewed
• [Journal Article] Subversion of Host Membrane Dynamics by the Legionella Dot/Icm TypeIV Secretion System. (2017)
  • Author(s): Hilbi H, Nagai H, Kubori T, Roy CR.
  • Journal Title: Curr Top Microbiol Immunol. Volume: 413 Pages: 221-242
  • DOI: 10.1007/978-3-319-75241-9_9
  • ISBN: 9783319752402, 9783319752419
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] Legionella RavZ Plays a Role in Preventing Ubiquitin Recruitment to Bacteria-Containing Vacuoles. (2017)
  • Author(s): Kubori T, Bui XT, Hubber A and Nagai H.
  • Journal Title: Front Cell Infect Microbiol. Volume: 7 Pages: 384-384
  • DOI: 10.3389/fcimb.2017.00384
  • Peer Reviewed / Open Access
• [Journal Article] Bacterial secretion system skews the fate of Legionella-containing vacuoles towards LC3-associated phagocytosis. (2017)
  • Author(s): Hubber A., Kubori T., Coban C., Matsuzawa T., Ogawa M., Kawabata T., Yoshimori T., and Nagai H.
  • Journal Title: Sci Rep. Volume: 7 Issue: 1 Pages: 44795-44795
  • DOI: 10.1038/srep44795
  • Peer Reviewed / Open Access / Int'l Joint Research / Acknowledgement Compliant
• [Journal Article] Isolation of the Dot/Icm Type IV Secretion System Core Complex from Legionella pneumophila for Negative Stain Electron Microscopy Studies (2017)
  • Author(s): Kubori T and Nagai H.
  • Journal Title: bio-protocol Volume: 7
  • Peer Reviewed / Open Access
• [Journal Article] Life with bacterial secretion systems. (2016)
  • Author(s): Kubori T.
  • Journal Title: PLoS Pathog. Volume: 12(8) Issue: 8 Pages: e1005562-e1005562
  • DOI: 10.1371/journal.ppat.1005562
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Presentation] Manipulation of the host ubiquitin system by Legionella (2019)
  • Author(s): Tomoko Kubori
  • Organizer: Cold Spring Harbor Asia Conference on Bacterial Infection and Host Defense
  • Int'l Joint Research / Invited
• [Presentation] Manipulation of the host ubiquitin system by Legionella (2018)
  • Author(s): Tomoko Kubori
  • Organizer: ICISE, Qui Nhon, Vietnam
  • Int'l Joint Research / Invited
• [Presentation] PLOS Pathogens Writing Workshop (2018)
  • Author(s): Tomoko Kubori
  • Organizer: ICISE, Qui Nhon, Vietnam
  • Int'l Joint Research / Invited
• [Presentation] Legionella manipulates host cellular systems utilizing the type IV secretion system (2018)
  • Author(s): 久堀智子
  • Organizer: 第 9１ 回日本細菌学会総会
• [Presentation] 宿主オートファジー関連システムとレジオネラ (2017)
  • Author(s): 久堀智子、Andree Hubber、Xuan Thanh Bui、永井宏樹
  • Organizer: 第 90 回日本細菌学会総会
  • Place of Presentation: 仙台国際センター展示棟（宮城県、仙台市）
  • Year and Date: 2017-03-19
• [Presentation] Manipulation of the host ubiquitin system by Legionella Deubiquitinases. (2017)
  • Author(s): Tomoko Kubori
  • Organizer: OIST workshop: Bacterial Flagella, Injectisome and Type III Secretion Systems
  • Place of Presentation: 沖縄科学技術大学院大学 Main Campus （沖縄県、国頭郡恩納村)
  • Year and Date: 2017-03-01
  • Int'l Joint Research
• [Presentation] 病原細菌レジオネラによる宿主ユビキチンシステムの操作 (2017)
  • Author(s): 久堀智子
  • Organizer: 新学術領域「運動超分子マシナリーが織りなす調和と多様性」 生体運動研究合同班会議
  • Place of Presentation: 神戸国際会議場 国際会議室 （兵庫県、神戸市）
  • Year and Date: 2017-01-06
• [Presentation] Manipulation of the host ubiquitin system by Legionella. (2017)
  • Author(s): Tomoko Kubori
  • Organizer: International Union of Microbiological Societies (IUMS2017)
  • Int'l Joint Research / Invited
• [Presentation] 宿主オートファジー関連システムとレジオネラ (2017)
  • Author(s): 久堀智子
  • Organizer: 日本顕微鏡学会 第 60 回記念シンポジウム
  • Invited
• [Presentation] Manipulation of the host ubiquitin system by Legionella Deubiquitinases. (2016)
  • Author(s): Tomoko Kubori and Hiroki Nagai
  • Organizer: T4SS2016: Type IV secretion in Gram-Nagative and Gram-Positive Bacteria
  • Place of Presentation: Beilngries, GERMANY
  • Year and Date: 2016-12-08
  • Int'l Joint Research
• [Presentation] 病原因子分泌系を介した細菌と宿主細胞の相互作用 (2016)
  • Author(s): 久堀智子
  • Organizer: 東京工業大学 Cell Biology Unit 第４回コロキウム
  • Place of Presentation: 東京工業大学すずかけ台キャンパス（神奈川県、横浜市）
  • Year and Date: 2016-10-21
  • Invited
• [Presentation] Manipulation of the host ubiquitin system by Legionella Deubiquitinases. (2016)
  • Author(s): Tomoko Kubori and Hiroki Nagai
  • Organizer: 第 15 回 あわじしま感染症・免疫フォーラム
  • Place of Presentation: 淡路夢舞台国際会議場（兵庫県、淡路市）
  • Year and Date: 2016-09-06
  • Int'l Joint Research
• [Presentation] Manipulation of the host ubiquitin system by Legionella Deubiquitinases. (2016)
  • Author(s): 久堀智子
  • Organizer: 新学術領域「運動超分子マシナリーが織りなす調和と多様性」 第 4 回領域全体会議
  • Place of Presentation: 長崎大学良順会館（長崎県、長崎市）
  • Year and Date: 2016-06-08
• [Book] Legionella Methods and Protocols Second Edition (2019)
  • Author(s): Tomoko Kubori, Hiroki Nagai
  • Total Pages: 476
  • Publisher: Springer Nature
  • ISBN: 9781493990474
• [Remarks] Nagai lab
  • URL: https://sites.google.com/view/nagai-lab
• [Remarks] Nagai Lab
  • URL: https://sites.google.com/site/nagailabjp/
• [Remarks] Nagai Lab
  • URL: http://nagailab.biken.osaka-u.ac.jp/