Tn-seq analysis of Legionella pathogenicity

• Project/Area Number: 19H03470
• Research Category: Grant-in-Aid for Scientific Research (B)
• Allocation Type: Single-year Grants
• Section: 一般
• Review Section: Basic Section 49050:Bacteriology-related
• Research Institution: Gifu University
• Principal Investigator: Nagai Hiroki 岐阜大学, 大学院医学系研究科, 教授 (80222173)
• Project Period (FY): 2019-04-01 – 2022-03-31
• Project Status: Completed (Fiscal Year 2022)
• Budget Amount: ¥17,290,000 (Direct Cost: ¥13,300,000、Indirect Cost: ¥3,990,000); Fiscal Year 2021: ¥5,590,000 (Direct Cost: ¥4,300,000、Indirect Cost: ¥1,290,000); Fiscal Year 2020: ¥5,590,000 (Direct Cost: ¥4,300,000、Indirect Cost: ¥1,290,000); Fiscal Year 2019: ¥6,110,000 (Direct Cost: ¥4,700,000、Indirect Cost: ¥1,410,000)
• Keywords: レジオネラ / TnSeq / Tn-Seq

Outline of Research at the Start

病原菌が病気を起こすメカニズムの解明のための一つの方法は、病原性を失った変異株を分離して、そこで変異を起こした遺伝子にどのような機能があるかを明らかにすることです。本研究では、次世代シーケンサーを利用したトランスポゾン・シーケンシング(Tn-seq)という方法を活用して、ヒトに重篤な肺炎を引き起こすレジオネラの病原遺伝子を真に網羅的に探索し、既存の研究で見えて来なかった新たな細菌感染戦略に迫ります。

Outline of Final Research Achievements

We conducted systematic search for genes involved in intracellualr growth of Legionella pneumophila using Tn-seq, and identified numerous genes other than dot/icm required for intracelluar growth.

Academic Significance and Societal Importance of the Research Achievements

Tn-seq法を用いたスクリーニングは、IV型分泌系遺伝子が見出された遺伝学的スクリーニングを発展させたものと捉えることができるが、文字通り真に網羅的に行うことが可能である点で画期的であり、実際新たな細菌側感染戦略の一端を明らかにすることができた。本研究の手法は他の病原微生物にも適応可能であり、当該領域に波及効果をもたらすことが期待できる。

Research Products

• [Int'l Joint Research] KAIST(韓国)
• [Int'l Joint Research] Oregon Health & Science University(米国)
• [Int'l Joint Research] KAIST(韓国)
• [Int'l Joint Research] Washington University(米国)
• [Int'l Joint Research] Kyungpook National University/KAIST(韓国)
• [Int'l Joint Research] Washington University(米国)
• [Journal Article] Mechanism of Lys6 poly-ubiquitin specificity by the L. pneumophila deubiquitinase LotA. (2023)
  • Author(s): Warren GD, Kitao T, Franklin TG, Nguyen JV, Geurink PP, Kubori T, Nagai H, Pruneda JN.
  • Journal Title: Molecular Cell Volume: 83(1):105-120.e5. Issue: 1 Pages: 105-120
  • DOI: 10.1016/j.molcel.2022.11.022
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Bacterial usurpation of the <scp>OTU</scp> deubiquitinase fold (2023)
  • Author(s): Pruneda Jonathan N.、Nguyen Justine V.、Nagai Hiroki、Kubori Tomoko
  • Journal Title: The FEBS Journal Volume: (2023) Issue: 15 Pages: 1-1
  • DOI: 10.1111/febs.16725
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Reversible modification of mitochondrial ADP/ATP translocases by paired <i>Legionella</i> effector proteins (2022)
  • Author(s): Kubori Tomoko、Lee Junyup、Kim Hyunmin、Yamazaki Kohei、Nishikawa Masanari、Kitao Tomoe、Oh Byung-Ha、Nagai Hiroki
  • Journal Title: Proceedings of the National Academy of Sciences Volume: 119 Issue: 23
  • DOI: 10.1073/pnas.2122872119
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] Legionella Manipulates Non-canonical SNARE Pairing Using a Bacterial Deubiquitinase. (2020)
  • Author(s): Tomoe Kitao, Kyoichiro Taguchi, Shintaro Seto, Kohei Arasaki, Hiroki Ando, Hiroki Nagai, Tomoko Kubori
  • Journal Title: Cell Reports Volume: 32(10) Issue: 10 Pages: 108107-108107
  • DOI: 10.1016/j.celrep.2020.108107
  • Peer Reviewed / Open Access
• [Journal Article] Structural basis for effector protein recognition by the Dot/Icm Type IVB coupling protein complex (2020)
  • Author(s): Kim Hyunmin、Kubori Tomoko、Yamazaki Kohei、Kwak Mi-Jeong、Park Suk-Youl、Nagai Hiroki、Vogel Joseph P.、Oh Byung-Ha
  • Journal Title: Nature Communications Volume: 11 Issue: 1 Pages: 2623-2623
  • DOI: 10.1038/s41467-020-16397-0
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Divergence of Legionella Effectors Reversing Conventional and Unconventional Ubiquitination. (2020)
  • Author(s): Tomoe Kitao, Hiroki Nagai, Tomoko Kubori
  • Journal Title: Frontiers in Cellular and Infection Microbiology Volume: 10 Pages: 448-448
  • DOI: 10.3389/fcimb.2020.00448
  • Peer Reviewed / Open Access
• [Presentation] 病原菌の細胞内寄生の分子基盤とその進化 (2021)
  • Author(s): 永井宏樹
  • Organizer: 第49回東海乳酸菌研究会研究報告会
  • Invited
• [Presentation] 型破りな翻訳後修飾を介したレジオネラの宿主細胞内生存戦略 (2021)
  • Author(s): 北尾公英、永井宏樹、久堀智子
  • Organizer: 第94回日本細菌学会総会シンポジウム
  • Invited
• [Presentation] Vacuole manipulation by Legionella deubiqutinases (2020)
  • Author(s): 久堀智子、北尾公英、永井宏樹
  • Organizer: 第93回日本細菌学会総会シンポジウム
  • Invited
• [Presentation] 細菌の病原性制御システム (2020)
  • Author(s): 北尾公英、久堀智子、永井宏樹
  • Organizer: 第93回日本細菌学会総会シンポジウム
  • Invited
• [Presentation] 私の研究歴: 若い研究者にお伝えしたいこと (2020)
  • Author(s): 永井宏樹
  • Organizer: 第93回日本細菌学会総会シンポジウム
  • Invited
• [Presentation] 病原菌の細胞内寄生の分子基盤とその進化 (2020)
  • Author(s): 永井宏樹
  • Organizer: 第48回東海乳酸菌研究会研究報告会
  • Invited
• [Presentation] 宿主SNARE pairing を操作するレジオネラエフェクターの解析 (2019)
  • Author(s): 北尾公英、田口馨一郎、瀬戸真太郎、新崎恒平、安藤弘樹、永井宏樹、久堀智子
  • Organizer: 第56回日本細菌学会中部支部総会
• [Presentation] Legionella manipulates non-canonical SNARE pairing by the function of a bacterial deubiquitinase (2019)
  • Author(s): Tomoe Kitao, Kohei Arasaki, Shintaro Seto, Kyoichiro Taguchi, Hiroki Nagai, Tomoko Kubori
  • Organizer: Microbial Adhesion and Signal Transduction, Gordon Research Conference
  • Int'l Joint Research
• [Remarks] 研究室webページ
  • URL: https://sites.google.com/view/nagai-lab/home
• [Remarks] 研究室webページ https://sites.google
  • URL: https://sites.google.com/view/nagai-lab/home