Characterization of expression and adherent property of staphylococcal surface proteins associated with staphylococcus infectious diseases.

• Project/Area Number: 19590439
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Single-year Grants
• Section: 一般
• Research Field: Bacteriology (including Mycology)
• Research Institution: National Institute of Infectious Diseases
• Principal Investigator: KURODA Makoto National Institute of Infectious Diseases, 病原体ゲノム解析研究センター・第三室, 室長 (80317411)
• Co-Investigator(Renkei-kenkyūsha): 田中 良和 北海道大学, 創成科学共同研究機構・北大基礎融合科学領域リーダー育成システム, 特任助教
• Project Period (FY): 2007 – 2008
• Project Status: Completed (Fiscal Year 2008)
• Budget Amount: ¥4,550,000 (Direct Cost: ¥3,500,000、Indirect Cost: ¥1,050,000); Fiscal Year 2008: ¥2,210,000 (Direct Cost: ¥1,700,000、Indirect Cost: ¥510,000); Fiscal Year 2007: ¥2,340,000 (Direct Cost: ¥1,800,000、Indirect Cost: ¥540,000)
• Keywords: 病原性 / 黄色ブドウ球菌 / 細胞壁架橋型蛋白質 / sarファミリー / 菌凝集 / バイオフィルム / SasG / Ebh

Research Abstract

ワクチン等による病原細菌の感染制御には初期感染メカニズムをより重点的に解明していく必要があり、本研究では定着因子候補である因子群とそれら制御レギュレーターの転写変動に着目した。黄色ブドウ球菌の21種の定着因子候補と8種の病原因子レギュレーターのマウス生体内での発現プロファイルを解析し、定着因子群がマウス生体内で強く転写増強していることを明らかにした。加えて、感染時においてブドウ球菌属に特徴的なSar 病原因子レギュレーターが転写増強し、それら制御下にあるsasG遺伝子が転写増強していることも明らかにした。sasG遺伝子に変異を導入すると菌凝集・バイオフィルム形成が消失し、プラスミドによりsasG遺伝子を相補すると回復した。SasGのAドメインの組換え発現体をNative電気泳動法およびフォルマリン架橋実験で分析したところ、4もしくは8量体のホモ複合体を形成していることが分かった。黄色ブドウ球菌の感染時において、Sarファミリーの複合的な転写調節系とSasGによる菌凝集・バイオフィルム形成が臨床で重要視される微小膿瘍形成に関与し、抗菌薬治療の難治化に関連している可能性が示唆された。

Research Products

• [Journal Article] Staphylococcus aureus Surface Protein SasG Contributes to Intercellular Autoaggregation of Staphylococcus aureus. (2008)
  • Author(s): Kuroda, M., Ito, R., Tanaka, Y., Yao, M., Matoba, K., Saito, S., Tanaka, I., Ohta, T.
  • Journal Title: Biochem Biophys Res Commun. 377(4) Pages: 1102-6
• [Journal Article] Structural basis for multimeric heme complexation through a specific protein-heme interaction: the case of the third neat domain of IsdH from Staphylococcus aureus. (2008)
  • Author(s): Watanabe, M., Tanaka, Y., Suenaga, A., Kuroda, M., Yao, M., Watanabe, N., Arisaka, F., Ohta, T., Tanaka, I., Tsumoto, K.
  • Journal Title: J Biol Chem. 283(42) Pages: 28649-59
• [Journal Article] Staphylococcus aureus giant protein Ebh is involved in tolerance to transient hyperosmotic pressure. (2008)
  • Author(s): Kuroda, M., Tanaka, Y., Aoki, R., Shu, D., Tsumoto, K., Ohta, T.
  • Journal Title: Biochem Biophys Res Commun. 374(2) Pages: 237-41
• [Journal Article] Expression, Purification, Crystallization, and Primary Diffraction Analysis of CapF, a Capsular Polysaccharide Synthesis Enzyme from Staphylococcus aureus. (2008)
  • Author(s): Miyafusa, T., Tanaka, Y., Kuroda, M., Ohta, T. and Tsumoto, K.
  • Journal Title: Acta Crystallogr Sect F Struct Biol Cryst Commun. 64 Pages: 512-5
• [Journal Article] A Helical String of Alternately Connected Two Three-helix Bundles for the 1.1-Megadalton Cell Wall-Associated Adhesion Protein Ebh from Staphylococcus aureus. (2008)
  • Author(s): Tanaka, Y., Sakamoto, S., Kuroda, M., Goda, S., Gao, YG., Tsumoto, K., Hiragi, Y., Yao, M., Watanabe, N., Ohta, T. and Tanaka, I.
  • Journal Title: Structure. 16 Pages: 488-496
• [Journal Article] Staphylococcus aureus Surface Protein SasG Contributes to Intercellular Autoaggregation of Staphylococcus aureus (2008)
  • Author(s): Kuroda, M., Ito, R., Tanaka, Y., Yao, M., Matoba, K., Saito, S., Tanaka, I., Ohta, T.
  • Journal Title: Biochem Biophys Res Commun. 377 Pages: 1102-1106
  • Peer Reviewed
• [Journal Article] Structural basis for multimeric heme complexation through a specific protein-heme interaction : the case of the third neat domain of IsdH from Staphylococcus aureus (2008)
  • Author(s): Watanabe, M., Tanaka, Y., Suenaga, A., Kuroda, M., Yao, M., Watanabe, N., Arisaka, F., Ohta, T., Tanaka, I., Tsumoto, K.
  • Journal Title: J Biol Chem. 283 Pages: 28649-28659
  • Peer Reviewed
• [Journal Article] Staphylococcus aureus giant protein Ebh is involved in tolerance to transient hyperosmotic pressure (2008)
  • Author(s): Kuroda, M., Tanaka, Y., Aoki, R., Shu, D., Tsumoto, K., Ohta, T.
  • Journal Title: Biochem Biophys Res Commun. 374 Pages: 237-241
  • Peer Reviewed
• [Journal Article] Expression, Purification, Crystallization, and Primary Diffraction Analysis of CapF, a Capsular Polysaccharide Synthesis Enzyme from Staphylococcus aureus (2008)
  • Author(s): Miyafusa, T., Tanaka, Y., Kuroda, M., Ohta, T., Tsumoto, K.
  • Journal Title: Acta Crystallogr Sect F Struct Biol Cryst Commun. 64 Pages: 512-515
  • Peer Reviewed
• [Journal Article] A helical string of alternately connected three-helix bundles for the cell wall-associated adhesion protein Ebh from Staphylococcus aureus. (2008)
  • Author(s): Yoshikazu Tanaka
  • Journal Title: Structure 16
  • Peer Reviewed
• [Journal Article] Crystal structure analysis reveals a novel forkhead-associated domain of ESAT-6 secretion system C protein in Staphylococcus aureus. (2007)
  • Author(s): Yoshikazu Tanaka
  • Journal Title: Proteins 69
  • Peer Reviewed
• [Presentation] 黄色ブドウ球菌のSarAパラログによる細胞壁架橋型タンパク質SasGの転写制御の解析 (2009)
  • Author(s): 黒田誠, 関塚剛史
  • Organizer: 第82回日本細菌学会総会
  • Place of Presentation: 名古屋国際会議場
  • Year and Date: 2009-03-12
• [Presentation] 黄色ブドウ球菌のSarAパラログによる細胞壁架橋型タンパク質SasGの転写制御の解析 (2009)
  • Author(s): 黒田誠、関塚剛史
  • Organizer: 第82回日本細菌学会
  • Place of Presentation: 名古屋国際会議場
• [Presentation] Methicillin-Resistant Staphylococcus saprophyticus Carrying Staphylococcal Cassette Chromosome mec (SCCmec) Have Emerged in Urogenital Tract Infections (2008)
  • Author(s): Makoto Kuroda, Masato Higashide, Carlos Takashi Neves Omura, Saburo Ohkawa, Sadahiro Ichimura, Toshiko Ohta.
  • Organizer: 108^<th> ASM general meeting.
  • Place of Presentation: Boston, MA USA.
• [Presentation] Methicillin-Resistant Staphylococcus saprophyticus Carrying Staphylococcal Cassette Chromosome mec (SCCmec) Have Emerged in Urogenital Tract Infections (2008)
  • Author(s): Makoto Kuroda, Masato Higashide, Carlos Takashi Neves Omura, Saburo Ohkawa, Sadahiro Ichimura, Toshiko Ohta
  • Organizer: 108^<th> ASM general meeting
  • Place of Presentation: Boston, MA USA.
• [Presentation] In vivo expression of cell surface proteins in Staphylococcus aureus. (2007)
  • Author(s): Makoto Kuroda
  • Organizer: Gordon Research Conference, staphylococcal diseases.
  • Place of Presentation: Les Diablerets,Switzerland
  • Year and Date: 2007-09-05
• [Presentation] In vivo expression of cell surface proteins in Staphylococcus aureus. (2007)
  • Author(s): Makoto Kuroda.
  • Organizer: Gordon Research Conference, staphylococcal diseases.
  • Place of Presentation: Sep. Les Diablerets, Switzerland
• [Presentation] Sesquiterpene Farnesol Affects Lipase Activity of Staphylococcus aureus as a Competitive Inhibitor. (2007)
  • Author(s): Makoto Kuroda, Sanae Nagasaki and Toshiko Ohta.
  • Organizer: 107^<th> ASM general meeting.
  • Place of Presentation: Toronto, Canada.
• [Presentation] Staphylococcus aureus surface protein SasG contributes to intercellular adhesion and biofilm formation. (2007)
  • Author(s): Ryuta Ito, Makoto Kuroda and Toshiko Ohta.
  • Organizer: 107^<th> ASM general meeting.
  • Place of Presentation: Toronto, Canada.
• [Presentation] (W5. 近縁ゲノム配列比較から病原性へ)比較ゲノム解析から見えるブドウ球菌属の種特徴的な病原性と生存機構 (2007)
  • Author(s): 黒田誠
  • Organizer: 第80回日本細菌学会総会
  • Place of Presentation: 大阪市アジア太平洋トレードセンター