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HilD is an AraC-like transcriptional regulator encoded in the  Salmonella  pathogenicity island 1 (SPI-1), which actives transcription of many genes within and outside SPI-1 that are mainly required for invasion of  Salmonella  into host cells. HilD controls expression of target genes directly or by acting through distinct regulators; three different regulatory cascades headed by HilD have been described to date. Here, by analyzing the effect of HilD on the  yobH  gene in  Salmonella enterica  serovar Typhimurium ( S . Typhimurium), we further define an additional regulatory cascade mediated by HilD, which was revealed by previous genome-wide analyses. In this regulatory cascade, HilD acts through SprB, a LuxR-like regulator encoded in SPI-1, to induce expression of virulence genes. Our data show that HilD induces expression of  sprB  by directly counteracting H-NS-mediated repression on the promoter region upstream of this gene. Then, SprB directly activates expression of several genes including  yobH ,  slrP  and  ugtL . Interestingly, we found that YobH, a protein of only 79 amino acids, is required for invasion of  S . Typhimurium into HeLa cells and mouse macrophages. Thus, our results reveal a novel  S . Typhimurium invasion factor and provide more evidence supporting the HilD-SprB regulatory cascade.

HilD induces expression of a novel Salmonella Typhimurium invasion factor, YobH, through a regulatory cascade involving SprB