Dual regulation of secretion system of periodontal virulence factors by novel two-component system and ECF sigma factor

• Project/Area Number: 22659332
• Research Category: Grant-in-Aid for Challenging Exploratory Research
• Allocation Type: Single-year Grants
• Research Field: Morphological basic dentistry
• Research Institution: Nagasaki University
• Principal Investigator: NAKAYAMA Koji 長崎大学, 大学院・医歯薬学総合研究科, 教授 (80150473)
• Co-Investigator(Renkei-kenkyūsha): TSUKUBA Tomoko (KADOWAKI Tomoko) 長崎大学, 大学院・医歯薬学総合研究科, 助教 (70336080) ; NAITO Mariko 長崎大学, 大学院・医歯薬学総合研究科, 准教授 (20244072) ; SHOJI Mikio 長崎大学, 大学院・医歯薬学総合研究科, 助教 (10336175) ; SATO Keiko 長崎大学, 大学院・医歯薬学総合研究科, 助教 (70410579) ; YUKITAKE Hideharu 長崎大学, 大学院・医歯薬学総合研究科, 技術職員 (30380984)
• Project Period (FY): 2010 – 2011
• Project Status: Completed (Fiscal Year 2011)
• Budget Amount: ¥3,140,000 (Direct Cost: ¥2,900,000、Indirect Cost: ¥240,000); Fiscal Year 2011: ¥1,040,000 (Direct Cost: ¥800,000、Indirect Cost: ¥240,000); Fiscal Year 2010: ¥2,100,000 (Direct Cost: ¥2,100,000)
• Keywords: 遺伝子 / 発現調節 / 二成分制御系 / シグマ因子 / 歯周病 / 歯学 / 細菌 / プロテアーゼ / 膜輸送

Research Abstract

We investigated regulatory mechanism of secretion system of pathogenic proteases/ adhesins of the periodontopathogen Porphyromonas gingivalis. Secretion of P. gingivalis gingipain proteases/ adhesins depends on a protein secretion system, Por secretion system(PorSS). We isolated 12 gingipain secretion-deficient mutants and found that two of them were deficient in a two-component system, which is a prokaryotic signaling system and consists of the sensor histidine kinase PorY and response regulator PorX. In this study, we constructed recombinant PorY and PorX proteins and found that PorY was autophosphorylated and the phosphoryl residue of PorY was transferred to PorX. PorY could bind to PorX as revealed by Surface plasmon resonance(BIACORE) analysis. Gene expression of component proteins of the PorSS was also regulated by one of ECF sigma factors, PGN0274, but the PorY-PorX system and the PGN0274 sigma factor independently regulate gene expression of component proteins of the PorSS, suggesting that the PorSS is regulated by multiple regulatory mechanisms.

Research Products

• [Journal Article] Suppression of RANKL-dependent heme oxygenase-1 is required for high mobility group box 1 release and osteoclastogenesis (2012)
  • Author(s): Sakai E, Shimada-Sugawara M, Nishishita K, Fukuma Y, Naito M, Okamoto K, Nakayama K, Tsukuba T
  • Journal Title: J Cell Biochem Volume: 113 Pages: 486-498
• [Journal Article] Por secretion system-dependent secretion and glycosylation of Porphyromonas gingivalis hemin-binding protein 35 (2011)
  • Author(s): Shoji M, Sato K, Yukitake H, Kondo Y, Narita Y, Kadowaki T, Naito M, and Nakayama K.
  • Journal Title: PLoS One Volume: 6
  • NAID: 120006985054
• [Journal Article] Characterisation of the Porphyromonas gingivalis conjugative transposon CTnPg1 : determination of the integration site and the genes essential for conjugal transfer (2011)
  • Author(s): Naito M, Sato K, Shoji M, Yukitake H, Ogura Y, Hayashi T and Nakayama K
  • Journal Title: Microbiology-SGM Volume: 157 Pages: 2022-2032
• [Journal Article] Effects of non-iron metalloporphyrins on growth and gene expression of Porphyromonas gingivalis (2011)
  • Author(s): Yukitake H, Naito M, Sato K, Shoji M, Ohara N, Yoshimura M, Sakai E and Nakayama K.
  • Journal Title: Microbiol Immunol Volume: 55 Pages: 141-153
  • NAID: 10031121453
• [Journal Article] Por secretion system-dependent secretion and glycosylation of Porphyromonas gingivalis hemin-binding protein 35 (2011)
  • Author(s): Shoji M, et al
  • Journal Title: PLoS One Volume: 6
  • NAID: 120006985054
  • Peer Reviewed
• [Journal Article] Effects of non-iron metalloporphyrins on growth and gene expression of Porphyromonas gingivalis. (2011)
  • Author(s): Yukitake H, Naito M, Sato K, Shoji M, Ohara N, Yoshimura M, Sakai E, Nakayama K
  • Journal Title: Microbiology and Immunology Volume: 55 Pages: 141-153
  • NAID: 10031121453
  • Peer Reviewed
• [Journal Article] A protein secretion system linked to bacteroidete gliding motility and pathogenesis (2010)
  • Author(s): Sato K, Naito M, Yukitake H, Hirakawa H, Shoji M, McBride MJ, Rhodes RG, and Nakayama K
  • Journal Title: Proc Natl Acad Sci USA Volume: 107 Pages: 276-281
  • NAID: 120006984618
• [Journal Article] Porphyromonas gingivalis peptidoglycans induce excessive activation of the innate immune system in silkworm larvae (2010)
  • Author(s): Ishii K, Hamamoto H, Imamura K, Adachi T, Shoji M, Nakayama K, and Sekimizu K.
  • Journal Title: J Biol Chem Volume: 285 Pages: 33338-33347
• [Journal Article] Porphyromonas gingivalis mutant defective in a putative glycosyltransferase exhibits defective biosynthesis of polysaccharide portions of lipopolysaccharide, decreased gingipain activities, strong autoaggregation and increased biofilm formation (2010)
  • Author(s): Yamaguchi M, Sato K, Yukitake H, Noiri Y, Ebisu S, and Nakayama K
  • Journal Title: Infect Immun Volume: 78 Pages: 3801-3812
• [Journal Article] Tetratricopeptide repeat protein-associated proteins contribute to the virulence of Porphyromonas gingivalis (2010)
  • Author(s): Kondo Y, Ohara N, Sato K, Yoshimura M, Yukitake H, Naito M, Fujiwara T, and Nakayama K
  • Journal Title: Infect Immun Volume: 78 Pages: 2846-2856
  • NAID: 120006984317
• [Journal Article] Characterization of hemin-binding protein 35(HBP35) in Porphyromonas gingivalis : its cellular distribution, thioredoxin activity and role in heme utilization (2010)
  • Author(s): Shoji M, Shibata Y, Shiroza T, Yukitake H, Peng B, Chen Y-Y, Sato K, Naito M, Abiko Y, Reynolds EC, and Nakayama K
  • Journal Title: BMC Microbiol Volume: 10 Pages: 152-152
• [Journal Article] Neutralization of toxic haem by Porphyromonas gingivalis haemoglobin receptor (2010)
  • Author(s): Nhien NT, Huy NT, Naito M, Oida T, Uyen DT, Huang M, Kikuchi M, Harada S, Nakayama K, Hirayama K, and Kamei K
  • Journal Title: J. Biochem Volume: 147 Pages: 317-325
  • NAID: 10027874258
• [Journal Article] Recombinant Porphyromonas gingivalis FimA preproprotein expressed in Escherichia coli is lipidated and a mature/ processed recombinant FimA protein forms a short filament in vitro (2010)
  • Author(s): Shoji M, Yoshimura A, Yoshioka H, Takade A, Takuma Y, Yukitake H, Naito M, Hara Y, Yoshida S, and Nakayama K
  • Journal Title: Can J Microbiol Volume: 56 Pages: 959-967
• [Journal Article] Hemmaglutinin/ adhesion domains of Porphyromonas gingivalis play key roles in coaggregation with Treponema denticola (2010)
  • Author(s): Ito R, Ishihara K, Shoji M, Nakayama K, and Okuda K.
  • Journal Title: FEMS Immunol Med Microbiol Volume: 60 Pages: 251-260
• [Journal Article] Porphyromonas gingivalis cell-induced hemagglutination and platelet aggregation (2010)
  • Author(s): Nakayama K
  • Journal Title: Periodontol 2000 Volume: 54 Pages: 45-52
• [Journal Article] Hemmaglutinin/adhesion Domains of Porphyromonas gingivalis Play Key Roles in Coaggregation with Treponema denticola. (2010)
  • Author(s): Ito R, Ishihara K, Shoji M, Nakayama K, Okuda K
  • Journal Title: FEMS Immunol.Med.Microbiol. Volume: 60 Pages: 251-260
  • Peer Reviewed
• [Journal Article] Recombinant Porphyromonas gingivalis FimA preproprotein expressed in Escherichia coli is lipidated and a mature or processed recombinant FimA protein forms a short filament in vitro. (2010)
  • Author(s): Shoji M, Yoshimura A, Yoshioka H, Takade A, Takuma Y, Yukitake H, Naito M, Hara Y, Yoshida S, Nakayama K
  • Journal Title: Canadian Journal of Microbiology Volume: 56 Pages: 959-967
  • NAID: 120006985158
  • Peer Reviewed
• [Journal Article] Porphyromonas gingivalis peptidoglycans induce excessive activation of the innate immune system in silkworm larvae. (2010)
  • Author(s): Ishii K, Hamamoto H, Imamura K, Adachi T, Shoji M, Nakayama K, Sekimizu K
  • Journal Title: J.Biol.Chem. Volume: 285 Pages: 33338-33347
  • Peer Reviewed
• [Journal Article] Porphyromonas gingivalis cell-induced hemagglutination and platelet aggregation. (2010)
  • Author(s): Nakayama K
  • Journal Title: Periodontology 2000 Volume: 54 Pages: 45-52
  • Peer Reviewed
• [Journal Article] Porphyromonas gingivalis mutant defective in a putative glycosyltransferase exhibits defective biosynthesis of polysaccharide portions of lipopolysaccharide, decreased gingioain activities, strong autoaggregation and increased biofilm formation. (2010)
  • Author(s): Yamaguchi M, Sato K, Yukitake H, Noiri Y, Ebisu S, Nakayama K
  • Journal Title: Infection and Immunity Volume: 78 Pages: 3801-3812
  • Peer Reviewed
• [Journal Article] Tetratricopeptide repeat protein-associated proteins contribute to the virulence of Porphyromonas gingivalis. (2010)
  • Author(s): Kondo Y, Ohara N, Sato K, Yoshimura M, Yukitake H, Naito M, Fujiwara T, Nakayama K
  • Journal Title: Infection and Immunity Volume: 78 Pages: 2846-2856
  • NAID: 120006984317
  • Peer Reviewed
• [Journal Article] Characterization of hemin-binding protein 35 (HBP35) in Porphyromonas gingivalis : its cellular distribution, thioredoxin activity and role in heme utilization (2010)
  • Author(s): Shoji M, Shibata Y, Shiroza T, Yukitake H, Pang B, Chen Y-Y, Sato K, Naito M, Abiko Y, Reynolds EC, Nakayama K
  • Journal Title: BMC Microbiology Volume: 10(online, journal) Pages: 152-152
  • Peer Reviewed
• [Journal Article] Neutralization of toxic haem by Porphyromonas gingivalis haemoglobin receptor. (2010)
  • Author(s): Nhien NT, Huy NT, Naito M, Oida T, Uyen DT, Huang M, Kikuchi M, Harada S, Nakayama K, Hirayama K, Kamei K
  • Journal Title: J.Biochem Volume: 147 Pages: 317-325
  • NAID: 10027874258
  • Peer Reviewed
• [Presentation] A novel protein secretion system, Por secretion system in periodontal pathogens (2011)
  • Author(s): Nakayama K
  • Organizer: International Union of Microbiological Societies 2011
  • Place of Presentation: Congress in Sapporo
• [Presentation] バクテロイデーテス細菌におけるタンパク分泌、滑走運動および病原性 (2010)
  • Author(s): 佐藤啓子, 内藤真理子, 雪竹英治, 庄子幹郎, 中山浩次
  • Organizer: 歯科基礎医学会
  • Place of Presentation: 東京
  • Year and Date: 2010-09-22
• [Presentation] Genome analysis and a novel secretion system of the periodontal pathogen Porphyromonas gingivalis (2010)
  • Author(s): Nakayama K
  • Organizer: 日本学術振興会・中国国家自然科学基金委員会主催第3回日中科学フォーラム
  • Place of Presentation: 武漢,中国
• [Presentation] 比較ゲノム解析から発見されたグラム陰性細菌の新規タンパク分泌機構 (2010)
  • Author(s): 中山浩次
  • Organizer: 第4回日本ゲノム微生物学会年会
  • Place of Presentation: 福岡
• [Patent(Industrial Property Rights)] 口腔細菌の迅速検出法 (2011)
  • Inventor(s): 吉村満美子、中山浩次、大原直也、竹原直道、吉田明弘
  • Industrial Property Rights Holder: 国立大学法人長崎大学
  • Acquisition Date: 2011-08-05