The response regulator SsrB activates expression of diverse Salmonella pathogenicity island 2 promoters and counters silencing by the nucleoid-associated protein H-NS

Don Walthers, Ronan K. Carroll, William Wiley Navarre, Stephen J. Libby, Ferric C. Fang, Linda Kenney

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Abstract

The two-component system SsrA-SsrB activates expression of a type III secretion system required for replication in macrophages and systemic infection in mice. Here we characterize the SsrB-dependent regulation of genes within Salmonella pathogenicity island 2 (SPI-2). Primer extension and DNase I footprinting identified multiple SsrB-regulated promoters within SPI-2 located upstream of ssaB, sseA, ssaG and ssaM. We previously demonstrated that ssrA and ssrB transcription is uncoupled. Overexpression of SsrB in the absence of its cognate kinase, SsrA, is sufficient to activate SPI-2 transcription. Because SsrB requires phosphorylation to relieve inhibitory contacts that occlude its DNA-binding domain, additional components must phosphorylate SsrB. SPI-2 promoters examined in single copy were highly SsrB-dependent, activated during growth in macrophages and induced by acidic pH. The nucleoid structuring protein H-NS represses horizontally acquired genes; we confirmed that H-NS is a negative regulator of SPI-2 gene expression. In the absence of H-NS, the requirement for SsrB in activating SPI-2 genes is substantially reduced, suggesting a role for SsrB in countering H-NS silencing. SsrB activates transcription of multiple operons within SPI-2 by binding to degenerate DNA targets at diversely organized promoters. SsrB appears to possess dual activities to promote SPI-2 gene expression: activation of transcription and relief of H-NS-mediated repression.

Original languageEnglish (US)
Pages (from-to)477-493
Number of pages17
JournalMolecular Microbiology
Volume65
Issue number2
DOIs
StatePublished - Jul 1 2007
Externally publishedYes

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Genomic Islands
Salmonella
Proteins
Macrophages
Genes
Gene Expression
Deoxyribonuclease I
DNA
Operon
Transcriptional Activation
Phosphotransferases
Phosphorylation

ASJC Scopus subject areas

  • Molecular Biology
  • Microbiology

Cite this

The response regulator SsrB activates expression of diverse Salmonella pathogenicity island 2 promoters and counters silencing by the nucleoid-associated protein H-NS. / Walthers, Don; Carroll, Ronan K.; Navarre, William Wiley; Libby, Stephen J.; Fang, Ferric C.; Kenney, Linda.

In: Molecular Microbiology, Vol. 65, No. 2, 01.07.2007, p. 477-493.

Research output: Contribution to journalArticle

Walthers, Don ; Carroll, Ronan K. ; Navarre, William Wiley ; Libby, Stephen J. ; Fang, Ferric C. ; Kenney, Linda. / The response regulator SsrB activates expression of diverse Salmonella pathogenicity island 2 promoters and counters silencing by the nucleoid-associated protein H-NS. In: Molecular Microbiology. 2007 ; Vol. 65, No. 2. pp. 477-493.
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