Genome-wide search for genes of Type IX secretion system by effective Tn-Seq analysis

• Project/Area Number: 17H04370
• Research Category: Grant-in-Aid for Scientific Research (B)
• Allocation Type: Single-year Grants
• Section: 一般
• Research Field: Morphological basic dentistry
• Research Institution: Nagasaki University
• Principal Investigator: NAITO Mariko 長崎大学, 医歯薬学総合研究科(歯学系), 教授 (20244072)
• Co-Investigator(Kenkyū-buntansha): 坂井 詠子 長崎大学, 医歯薬学総合研究科(歯学系), 助教 (10176612)
• Project Period (FY): 2017-04-01 – 2020-03-31
• Project Status: Completed (Fiscal Year 2019)
• Budget Amount: ¥17,030,000 (Direct Cost: ¥13,100,000、Indirect Cost: ¥3,930,000); Fiscal Year 2019: ¥4,420,000 (Direct Cost: ¥3,400,000、Indirect Cost: ¥1,020,000); Fiscal Year 2018: ¥3,640,000 (Direct Cost: ¥2,800,000、Indirect Cost: ¥840,000); Fiscal Year 2017: ¥8,970,000 (Direct Cost: ¥6,900,000、Indirect Cost: ¥2,070,000)
• Keywords: 歯周病原菌 / ９型分泌機構 / Tn-Seq / 9型分泌機構 / 細菌 / 口腔細菌

Outline of Final Research Achievements

Genes related to T9SS were comprehensively searched by constructing a high-density Tn insertion mutant library of Porphyromonas gingivalis. As a result, it was revealed that the PorG gene is a new essential gene of T9SS. One of previously reported T9SS related gene, GPPX, showed no relation with T9SS. Furthermore, we identified the genes involved in lipid modification of T9SS cargo proteins and analyzed their functional roles. We also clarified the function of a common structure (Ig-like domain) inside the T9SS transport protein.

Academic Significance and Societal Importance of the Research Achievements

Porphyromonas gingivalisは現在でもほとんどの日本人の歯牙を失う原因である歯周病の主要な原因菌である。本研究では本菌の多様な病原因子の菌体外への輸送に働く輸送機構（９型分泌機構）の全容を把握するために、網羅的に必須遺伝子の同定を行った。また同定した必須遺伝子を機能でグループ分けを行うことができた。９型分泌機構は他の菌属には見られない特殊な機構である。その為、本研究の成果は歯周病原菌を標的にした新たな創薬開発に役立つ情報基盤として意義深いと考えている。

Research Products

• [Journal Article] Tree of motility;A proposed history of motility systems in the tree of life (2020)
  • Author(s): M Miyata, R C Robinson, T QP Uyeda, Y Fukumori, S Fukushima, S Haruta, M Homma, K Inaba, M Ito, C Kaito, K Kato, TKenri, Y Kinosita, S Kojima, T Minamino, H Mori, S Nakamura, D Nakane, K Nakayama, M Nishiyama, S Shibata, K Shimabukuro, M Tamakoshi, A Taoka, Y Tashiro, ITulum, HiWada, K Wakabayashi
  • Journal Title: Genes to Cells Volume: 25 Issue: 1 Pages: 6-21
  • DOI: 10.1111/gtc.12737
  • NAID: 120006881482
  • Peer Reviewed / Open Access / Int'l Joint Research
• [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] PGN_0297 is an essential component of the type IX secretion system (T9SS) in Porphyromonas gingivalis: Tn-seq analysis for exhaustive identification of T9SS-related genes. (2019)
  • Author(s): Naito M, Tominaga T, Shoji M, Nakayama K.
  • Journal Title: Microbiol Immunol. Volume: 63(1) Issue: 1 Pages: 11-20
  • DOI: 10.1111/1348-0421.12665
  • Peer Reviewed / Open Access
• [Journal Article] Identification of genes encoding glycosyltransferases involved in lipopolysaccharide synthesis in Porphyromonas gingivalis. (2018)
  • Author(s): Shoji M, Sato K, Yukitake H, Kamaguchi A, Sasaki Y, Naito M, Nakayama K.
  • Journal Title: Mol Oral Microbiol. Volume: 33(1) Issue: 1 Pages: 68-80
  • DOI: 10.1111/omi.12200
  • Peer Reviewed / Open Access
• [Journal Article] Immunoglobulin-like domains of the cargo proteins are essential for protein stability during secretion by the type IX secretion system. (2018)
  • Author(s): Sato K, Kakuda S, Yukitake H, Kondo Y, Shoji M, Takebe K, Narita Y, Naito M, Nakane D, Abiko Y, Hiratsuka K, Suzuki M, Nakayama K.
  • Journal Title: Mol Microbiol. Volume: 110(1) Issue: 1 Pages: 64-81
  • DOI: 10.1111/mmi.14083
  • Peer Reviewed
• [Presentation] PGN_0297 (porG)はPorphyromonas gingivalisのIX型分泌機構の必須構成遺伝子である (2019)
  • Author(s): 内藤真理子, 庄子幹郎, 中山浩次
  • Organizer: 第92回日本細菌学会総会
• [Presentation] 9型分泌機構の基質タンパク質であるPorAは二成分制御系PorXYとECFシグマ因子SigPの上流で正の制御因子として機能する (2019)
  • Author(s): 庄子幹郎, 雪竹英治, 佐藤啓子, 内藤真理子, 中山浩次
  • Organizer: 第72回日本細菌学会九州支部総会・第56回日本ウイルス学会九州支部総会
• [Presentation] 9型分泌機構(T9SS)のカーゴタンパク質PorAは、二成分制御系PorXYおよびECFシグマ因子SigPの上流でT9SS発現の調節因子として作用する (2019)
  • Author(s): 中山浩次, 雪竹英治, 庄子幹郎, 佐藤啓子, 内藤真理子
  • Organizer: 第61回歯科基礎医学会学術大会
• [Presentation] PGN_0297(porG)はPorphyromonas gingivalisのIX型分泌機構の必須構成遺伝子である (2019)
  • Author(s): 内藤真理子, 庄子幹郎, 中山浩次
  • Organizer: 第61回歯科基礎医学会学術大会
• [Presentation] Glycosyltransferase-encoding genes involved in LPS synthesis in Porphyromonas gingivali (2018)
  • Author(s): 1.庄子幹郎，佐藤啓子，雪竹英治，内藤真理子，中山浩次
  • Organizer: 第91回日本細菌学会総会
• [Presentation] 高密度Tn-seq法によるIX型分泌機構関連遺伝子の探索 (2017)
  • Author(s): 内藤真理子(代表)，庄子幹郎，中山浩次
  • Organizer: 第70回日本細菌学会九州支部総会・第54回日本ウイルス学会九州支部総会