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PLoS One. 2011;6(8):e23314. doi: 10.1371/journal.pone.0023314. Epub 2011 Aug 18.

Phosphorylated CpxR restricts production of the RovA global regulator in Yersinia pseudotuberculosis.

Author information

1
Department of Molecular Biology, Umeå University, Umeå, Sweden.

Abstract

BACKGROUND:

RovA is a global transcriptional regulator of gene expression in pathogenic Yersinia. RovA levels are kept in check by a sophisticated layering of distinct transcriptional and post-transcriptional regulatory mechanisms. In the enteropathogen Y. pseudotuberculosis, we have previously reported that the extracytoplasmic stress sensing CpxA-CpxR two-component regulatory system modulates rovA expression.

METHODOLOGY/PRINCIPAL FINDINGS:

In this study, we characterized CpxR phosphorylation (CpxR∼P) in vitro, and determined that phosphorylation was necessary for CpxR to efficiently bind to the PCR-amplified upstream regulatory region of rovA. The precise CpxR∼P binding site was mapped by a nuclease protection assay and directed mutagenesis confirmed that in vivo binding to the rovA promoter inhibits transcription. Reduced RovA production was most pronounced following CpxR∼P accumulation in the Yersinia cytoplasm during chronic Cpx pathway activation and by the indiscriminate phosphodonor action of acetyl phosphate.

CONCLUSIONS/SIGNIFICANCE:

Cpx pathway activation restricts levels of the RovA global regulator. The regulatory influence of CpxR∼P must therefore extend well beyond periplasmic quality control in the Yersinia envelope, to include genes involved in environmental survival and pathogenicity.

PMID:
21876746
PMCID:
PMC3158067
DOI:
10.1371/journal.pone.0023314
[Indexed for MEDLINE]
Free PMC Article

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