Analysis of the Salmonella regulatory network suggests involvement of SsrB and H-NS in σ^E-regulated SPI-2 gene expression

The extracytoplasmic functioning sigma factor σ^E is known to play an essential role for Salmonella enterica serovar Typhimurium to survive and proliferate in macrophages and mice. However, its regulatory network is not well-characterized, especially during infection. Here we used microarray to identify genes regulated by σ^E in Salmonella grown in three conditions: a nutrient-rich condition and two others that mimic early and late intracellular infection. We found that in each condition σ^E regulated different sets of genes, and notably, several global regulators. When comparing nutrient-rich and infection-like conditions, large changes were observed in the expression of genes involved in Salmonella pathogenesis island (SPI)-1 type-three secretion system (TTSS), SPI-2 TTSS, protein synthesis, and stress responses. In total, the expression of 58% of Salmonella genes was affected by σ^E in at least one of the three conditions. An important finding is that σ^E up-regulates SPI-2 genes, which are essential for Salmonella intracellular survival, by up-regulating SPI-2 activator ssrB expression at the early stage of infection and down-regulating SPI-2 repressor hns expression at a later stage. Moreover, σ^E is capable of countering the silencing of H-NS, releasing the expression of SPI-2 genes. This connection between σ^E and SPI-2 genes, combined with the global regulatory effect of σ^E, may account for the lethality of rpoE-deficient Salmonella in murine infection.