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Background  The 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.    Results  Of the 366 examined spots, 269 proteins were successfully identified. The comparative analysis of the wild-type and ppGpp 0  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.    Conclusions  A 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 .

bmc microbiology

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