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A link between transcription termination, antibiotic action and virulence: Inhibition of Rho activity increases expression of SaeRS-dependent virulence factor genes in Staphylococcus aureus

ABSTRACT: Comparative proteome analysis of cells and supernatant from S. aureus HG001 and its isogenic rho deletion mutant in exponential and stationary growth phase in RPMI and TSB medium. Proteome analysis of supernatant from S. aureus HG001 and its isogenic rho deletion mutant in RPMI medium after treatment with Bicyclomycin (BCM).

INSTRUMENT(S): Q Exactive

ORGANISM(S): Staphylococcus Aureus Hg001

SUBMITTER: Uwe Voelker

PROVIDER: MSV000082270 | MassIVE | Thu Apr 12 08:46:00 BST 2018

REPOSITORIES: MassIVE

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Transcriptome analysis of a Staphylococcus aureus mutant defective in the transcription termination factor Rho.

Project description:This analysis is part of the study Whole-transcriptome analysis of Staphylococcus aureus under laboratory and infection-mimicking conditions (MÃ¤der, Nicolas et al., to be submitted) where the S. aureus HG001 transcriptome was analyzed under more than 40 different biological conditions. The data revealed a relatively low abundance of antisense RNAs in S. aureus, overlapping only 6% of the coding genes. Transcriptome analysis of the rho deletion mutant revealed a remarkable overall increase in antisense transcription in S. aureus. delta rho and wild type

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Project description:CodY is a conserved regulator in gram-positive organisms described to repress metabolic genes mainly involved in nitrogen metabolism but also to control the expression of virulence genes in pathogens. We constructed codY gene-replacement mutants in three unrelated S. aureus strains (Newman, UAMS-1, RN1HG). codY mutants grew slower in a chemically defined medium compared to the wild type strains. However, only codY mutants were able to grow in medium lacking threonine. Excess of isoleucine resulted in growth inhibition in the wild type but not in the codY mutants indicating a role of isoleucine for CodY dependent repression of metabolic genes. The prototypic CodY-repressed operon ilvDBCleuABCilvA is preceded by a CodY-binding motif and repressed after up-shift with isoleucine and to a lesser extent guanidine. Transcription of the quorum sensing system agr followed a similar expression pattern. The codY dependent repression of agr is in line with the concomitant influence of CodY on typical agr regulated genes such as cap, spa fnbA and coa. However, transcriptional analysis revealed that most of these virulence genes (e.g. cap, fnbA, spa hla) are also regulated by CodY in an agr negative background. Microarray analysis revealed the large majority of codY-repressed genes are involved in amino-acid transport and metabolism, genes showing codY dependent activation were mainly involved in nucleotide transport and metabolism or virulence. In summary, CodY in S. aureus not only acts as repressor for genes involved in nitrogen metabolism but also contributes to virulence gene regulation by supporting as well as substituting agr function.

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CodY in Staphylococcus aureus: regulatory link between metabolism and virulence gene expression

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