Elucidation of feedback regulation of the expression of the type IX secretion system (T9SS) of Porphyromonas gingivalis

• Project/Area Number: 20K09921
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Review Section: Basic Section 57020:Oral pathobiological science-related
• Research Institution: Nagasaki University
• Principal Investigator: NAKAYAMA Koji 長崎大学, 医歯薬学総合研究科(歯学系), 客員研究員 (80150473)
• Co-Investigator(Kenkyū-buntansha): 雪竹 英治 長崎大学, 医歯薬学総合研究科(歯学系), 技術職員 (30380984) ; 庄子 幹郎 長崎大学, 医歯薬学総合研究科(歯学系), 准教授 (10336175) ; 内藤 真理子 長崎大学, 医歯薬学総合研究科(歯学系), 教授 (20244072)
• Project Period (FY): 2020-04-01 – 2023-03-31
• Project Status: Completed (Fiscal Year 2022)
• Budget Amount: ¥4,420,000 (Direct Cost: ¥3,400,000、Indirect Cost: ¥1,020,000); Fiscal Year 2022: ¥1,430,000 (Direct Cost: ¥1,100,000、Indirect Cost: ¥330,000); Fiscal Year 2021: ¥1,430,000 (Direct Cost: ¥1,100,000、Indirect Cost: ¥330,000); Fiscal Year 2020: ¥1,560,000 (Direct Cost: ¥1,200,000、Indirect Cost: ¥360,000)
• Keywords: 細菌 / 病原性 / 歯学 / 歯周病 / 分泌 / 調節 / 生体分子 / 感染症

Outline of Research at the Start

病原細菌の多くは毒素、プロテアーゼなどの酵素、凝集素などを菌体表面および菌体外に分泌する。私たちは9型分泌機構（T9SS）を世界に先駆けて歯周病細菌ジンジバリス菌にて発見した。ジンジパインなどの本菌の重要な病原因子はT9SSで分泌される。T9SSは二成分制御系とECFシグマ因子の直列シグナル経路により発現調節されるが、調節系の全体像は不明であった。本研究は、このシグナル経路の上流に位置する新規の外膜タンパク質PorAによるフィードバック制御を解明することにあり、その成果は歯周病細菌を始めとするバクテロイデーテス門細菌のT9SSをコントロールする方法の開発という応用研究に発展する可能性がある。

Outline of Final Research Achievements

PorA involved in the expression of the type IX secretion system (T9SS) of the periodontopathogenic bacterium Porphyromonas gingivalis is a protein composed of a signal peptide, a FimH-like domain, and a CTD domain. The FimH-like domain was found to be irrelevant to the involvement of PorA in T9SS expression, and CTD[PorA] was the functional domain. CTD[PorA] was found to bind to the periplasmic domain of the PorY sensor protein. It was suggested that the amount of CTD[PorA] in the periplasm regulates the expression of T9SS component proteins through the PorA-PorY-PorX-SigP signaling pathway.

Academic Significance and Societal Importance of the Research Achievements

歯周病細菌ポルフィロモナス・ジンジバリスは病原性プロテアーゼであるジンジパインなどの病原因子をT9SSで菌体表面や菌体外に分泌する。歯周病の治療戦略として本細菌のT9SSを標的とする分子標的薬の開発が有効と思われる。T9SSの発現を調節する分子機構に関わるPorAタンパク質の詳細な解析は分子標的薬を構築する上でPorA分子内のどこを標的とする化合物がT9SSを制御する可能性があるかを示唆しており、分子標的薬開発に寄与すると考えられる。

Research Products

• [Int'l Joint Research] University of Wisconsin-Milwaukee(米国)
• [Journal Article] Filamentous structures in the cell envelope are associated with bacteroidetes gliding machinery (2023)
  • Author(s): Shibata Satoshi, Tahara Yuhei O., Katayama Eisaku, Kawamoto Akihiro, Kato Takayuki, Zhu Yongtao, Nakane Daisuke, Namba Keiichi, Miyata Makoto, McBride Mark J., Nakayama Koji
  • Journal Title: Communications Biology Volume: 6 Issue: 1 Pages: 94-94
  • DOI: 10.1038/s42003-023-04472-3
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Porphyromonas gingivalis Gingipains Induce Cyclooxygenase-2 Expression and Prostaglandin E2 Production via ERK1/2-Activated AP-1 (c-Jun/c-Fos) and IKK/NF-κB p65 Cascades (2022)
  • Author(s): Nakayama Masaaki、Naito Mariko、Omori Kazuhiro、Ono Shintaro、Nakayama Koji、Ohara Naoya
  • Journal Title: The Journal of Immunology Volume: 208 Issue: 5 Pages: 1146-1154
  • DOI: 10.4049/jimmunol.2100866
  • Peer Reviewed / Open Access
• [Journal Article] Insertional Inactivation and Gene Complementation of Prevotella intermedia Type IX Secretion System Reveals Its Indispensable Roles in Black Pigmentation, Hemagglutination, Protease Activity of Interpain A, and Biofilm Formation (2022)
  • Author(s): Naito M, Shoji M, Sato K, Nakayama K
  • Journal Title: Journal of Bacteriology Volume: 204(8) Issue: 8
  • DOI: 10.1128/jb.00203-22
  • Peer Reviewed
• [Journal Article] Colony spreading of the gliding bacterium Flavobacterium johnsoniae in the absence of the motility adhesin SprB (2021)
  • Author(s): Sato Keiko、Naya Masami、Hatano Yuri、Kondo Yoshio、Sato Mari、Narita Yuka、Nagano Keiji、Naito Mariko、Nakayama Koji、Sato Chikara
  • Journal Title: Scientific Reports Volume: 11 Issue: 1 Pages: 697-697
  • DOI: 10.1038/s41598-020-79762-5
  • NAID: 120006956183
  • Peer Reviewed / Open Access
• [Journal Article] Transport and Polymerization of Porphyromonas gingivalis Type V Pili (2020)
  • Author(s): Shoji M, Shibata S, Naito M, Nakayama K.
  • Journal Title: Methods in Molecular Microbiology Volume: 2210 Pages: 61-73
  • DOI: 10.1007/978-1-0716-0939-2_7
  • ISBN: 9781071609385, 9781071609392
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] PorA, a conserved C-terminal domain-containing protein, impacts the PorXY-SigP signaling of the type IX secretion system. (2020)
  • Author(s): Yukitake H, Shoji M, Sato K, Handa Y, Naito M, Imada K, Nakayama K.
  • Journal Title: Scientific Reports Volume: 10(1) Issue: 1 Pages: 21109-21109
  • DOI: 10.1038/s41598-020-77987-y
  • NAID: 120006980768
  • Peer Reviewed / Open Access
• [Journal Article] Structure of polymerized type V pilin reveals assembly mechanism involving protease-mediated strand exchange (2020)
  • Author(s): Satoshi Shibata, Mikio Shoji, Kodai Okada, Hideyuki Matsunami, Melissa Matthews, Katsumi Imada, Koji Nakayama and Matthias Wolf
  • Journal Title: Nature Microbiology Volume: － Issue: 6 Pages: 830-837
  • DOI: 10.1038/s41564-020-0705-1
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Type IX Secretion System Cargo Proteins Are Glycosylated at the C Terminus with a Novel Linking Sugar of the Wbp/Vim Pathway. (2020)
  • Author(s): Veith PD, Shoji M, O'Hair RAJ, Leeming MG, Nie S, Glew MD, Reid GE, Nakayama K, Reynolds EC.
  • Journal Title: mBio Volume: 11(5) Issue: 5
  • DOI: 10.1128/mbio.01497-20
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] PorM, a core component of bacterial type IX secretion system, forms a dimer with a unique kinked-rod shape (2020)
  • Author(s): Sato Keiko、Okada Kodai、Nakayama Koji、Imada Katsumi
  • Journal Title: Biochemical and Biophysical Research Communications Volume: 532 Issue: 1 Pages: 114-119
  • DOI: 10.1016/j.bbrc.2020.08.018
  • Peer Reviewed
• [Journal Article] Biogenesis of Type V pili. (2020)
  • Author(s): Shoji M, Shibata S, Sueyoshi T, Naito M, Nakayama K.
  • Journal Title: Microbiology and Immunology Volume: 64(10) Issue: 10 Pages: 643-656
  • DOI: 10.1111/1348-0421.12838
  • NAID: 120006988474
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Effect of Porphyromonas gingivalis infection on gut dysbiosis and resultant arthritis exacerbation in mouse model (2020)
  • Author(s): Hamamoto Yuta、Ouhara Kazuhisa、Munenaga Syuichi、Shoji Mikio、Ozawa Tatsuhiko、Hisatsune Jyunzo、Kado Isamu、Kajiya Mikihito、Matsuda Shinji、Kawai Toshihisa、Mizuno Noriyoshi、Fujita Tsuyoshi、Hirata Shintaro、Tanimoto Kotaro、Nakayama Koji、Kishi Hiroyuki、Sugiyama Eiji、Kurihara Hidemi
  • Journal Title: Arthritis Research & Therapy Volume: 22 Issue: 1 Pages: 249-249
  • DOI: 10.1186/s13075-020-02348-z
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Presentation] The T9SS cargo protein PorA binds to a sensor kinase PorY to regulate the T9SS gene regulation (2023)
  • Author(s): 伊藤李香, 雪竹英治, 庄子幹郎, 藤原卓, 中山浩次, 内藤真理子
  • Organizer: 第96回日本細菌学会総会
• [Presentation] Porphyromonas gingivalisジンジパインによるPLCを介したCOX-2発現とPGE2産生の分子機序 (2023)
  • Author(s): 中山真彰, 山口智之, 内藤真理子, 中山浩次, 大原直也
  • Organizer: 第96回日本細菌学会総会
• [Presentation] 9型分泌機構の遺伝子発現制御に関わるPorAタンパク質のシグナル活性化機構 (2022)
  • Author(s): 伊藤李香, 雪竹英治, 庄子幹郎, 田上直美, 藤原卓, 中山浩次, 内藤真理子
  • Organizer: 九州微生物フォーラム2022
• [Presentation] T9SS CTDタンパク質の一つであるPorAはT9SS構成タンパク質の遺伝子発現調節に関わる (2021)
  • Author(s): 雪竹英治、庄子幹郎、佐藤啓子、反田祐介、内藤真理子、今田勝巳、中山浩次
  • Organizer: 日本細菌学会