Comparative proteomic analysis of Salmonella enterica serovar Typhimurium ppGpp-deficient mutant to identify a novel virulence protein required for intracellular survival in macrophages

BMC Microbiology

Research Article

Takeshi Haneda¹  Tsuyoshi Miki¹  Yoshinori Kumagai¹  Mariko Sugimoto¹  Satomi Shimizu-Izumi¹  Nobuhiko Okada¹  Hirofumi Danbara¹  Yukie Yoshida-Ohta¹  Tadakazu Maeda²  Yoshio Kodera²  Masamichi Oh-Ishi² 

[1] Department of Microbiology, School of Pharmacy, Kitasato University, 5-9-1 Shirokane, 108-8641, Minato-ku, Tokyo, Japan;Laboratory of Biomolecular Dynamics, Department of Physics, Kitasato University School of Science, 1-15-1 Kitasato, Minami-ku, 252-0373, Sagamihara City, Kanagawa, Japan;

关键词: Protein Spot;    Stringent Response;    Comparative Proteomic Analysis;    Typhimurium Strain;    Competitive Index;   

DOI  :  10.1186/1471-2180-10-324

 received in 2010-09-01, accepted in 2010-12-21,  发布年份 2010

来源: Springer

【 摘 要 】

BackgroundThe global ppGpp-mediated stringent response in pathogenic bacteria plays an important role in the pathogenesis of bacterial infections. In Salmonella enterica serovar Typhimurium (S. Typhimurium), several genes, including virulence genes, are regulated by ppGpp when bacteria are under the stringent response. To understand the control of virulence genes by ppGpp in S. Typhimurium, agarose 2-dimensional electrophoresis (2-DE) combined with mass spectrometry was used and a comprehensive 2-DE reference map of amino acid-starved S. Typhimurium strain SH100, a derivative of ATCC 14028, was established.ResultsOf the 366 examined spots, 269 proteins were successfully identified. The comparative analysis of the wild-type and ppGpp0 mutant strains revealed 55 proteins, the expression patterns of which were affected by ppGpp. Using a mouse infection model, we further identified a novel virulence-associated factor, STM3169, from the ppGpp-regulated and Salmonella-specific proteins. In addition, Salmonella strains carrying mutations in the gene encoding STM3169 showed growth defects and impaired growth within macrophage-like RAW264.7 cells. Furthermore, we found that expression of stm3169 was controlled by ppGpp and SsrB, a response regulator of the two-component system located on Salmonella pathogenicity island 2.ConclusionsA proteomic approach using a 2-DE reference map can prove a powerful tool for analyzing virulence factors and the regulatory network involved in Salmonella pathogenesis. Our results also provide evidence of a global response mediated by ppGpp in S. enterica.

【 授权许可】

Unknown   
© Haneda et al; licensee BioMed Central Ltd. 2010. This article is published under license to BioMed Central Ltd. This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.