Dataset Information

Differential Regulation of CsrC and CsrB by CRP-cAMP in Salmonella enterica.

ABSTRACT: Post-transcriptional regulation mediated by regulatory small RNAs (sRNAs) has risen as a key player in fine-tuning gene expression in response to environmental stimuli. Here, we show that, in Salmonella enterica, the central metabolic regulator CRP-cAMP differentially regulates the sRNAs CsrB and CsrC in a growth phase-dependent manner. While CsrB expression remains unchanged during growth, CsrC displays a growth phase-dependent expression profile, being weakly expressed at the logarithmic growth phase and induced upon entry into stationary phase. We show that CRP-cAMP contributes to the expression pattern of CsrC by repressing its expression during the logarithmic growth phase. The CRP-cAMP mediated repression of CsrC is independent of SirA, a known transcriptional CsrB/CsrC activator. We further show that the sRNA Spot 42, which is derepressed in a Δcrp strain, upregulates CsrC during logarithmic growth. We propose a model where the growth-dependent regulation of CsrC is sustained by the CRP-cAMP-mediated repression of Spot 42. Together, our data point toward a differential regulation of the sRNAs CsrB and CsrC in response to environmental stimuli, leading to fine-tuning of gene expression via the sequestration of the RNA-binding protein CsrA.

SUBMITTER: El Mouali Y

PROVIDER: S-EPMC7591399 | biostudies-literature | 2020

REPOSITORIES: biostudies-literature

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Publications

Differential Regulation of CsrC and CsrB by CRP-cAMP in <i>Salmonella enterica</i>.

El Mouali Youssef Y Esteva-Martínez Guillem G García-Pedemonte David D Balsalobre Carlos C

Frontiers in microbiology 20201014

Post-transcriptional regulation mediated by regulatory small RNAs (sRNAs) has risen as a key player in fine-tuning gene expression in response to environmental stimuli. Here, we show that, in <i>Salmonella enterica</i>, the central metabolic regulator CRP-cAMP differentially regulates the sRNAs CsrB and CsrC in a growth phase-dependent manner. While CsrB expression remains unchanged during growth, CsrC displays a growth phase-dependent expression profile, being weakly expressed at the logarithmi ...[more]

PMID: 33162952

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Project description:Colonization of the intestinal tract and dissemination into deeper tissues by the enteric pathoÂgen Yersinia pseudotuberculosis demands expression of a special set of virulence factors important for the initiation and the persistence of the infection. In this study we demonstrate that many virulence-associated functions are coregulated with the carbohydrate metabolism. This link is mediated by the carbon storage regulator (Csr) system, including the regulatory RNAs CsrB and CsrC, and the cAMP receptor protein (Crp), which both control virulence gene expression in response to the nutrient composition of the medium. Here, we show that Crp regulates the synthesis of both Csr RNAs in an opposite manner. A loss of the crp gene resulted in a strong upregulation of CsrB synthesis, whereas CsrC levels were strongly reduced leading to downregulation of the viruÂlence regulator RovA. Switching of the Csr RNA involves Crp-mediated reÂpression of the response regulator UvrY which activates csrB transcription. To elucidate the regulatory links between virulence and carbon metabolism, we performed comparative metabolome, transÂÂÂcripÂtome and phenotypic microarray analyses and found that Crp promotes oxidative catabolism of many different carbon sources, whereas fermentative patterns of metabolism are favoured when crp is deleted. Mouse infection experiments further demonstrated that Crp is pivotal for a successÂful Y. pseudoÂtuberÂculosis infection. In summary, placement of the Csr system and important virulence factors under control of Crp enables this pathogen to link its nutritional status to virulence in order to optimize bioÂlogical fitness and infection efficiency through the infecÂtious life cycle. Y. pseudotuberculosis YPIII or the isogenic crp mutant strain were grown to late stationary phase at 25Â°C. Four biological replicates were employed for each experiment consisting of two pooled individual cultures and two pooled RNA preparation samples, respectively. Total RNA was extracted using SV Total RNA Isolation System (Promega). The samples were treated with RNase-free DNase (Roche Applied Science) and the quality of the RNA was confirmed by the lack of PCR amplification of the hns gene and by using an Agilent 2100 Bioanalyzer.

Dataset Information

Differential Regulation of CsrC and CsrB by CRP-cAMP in Salmonella enterica.

Publications

Differential Regulation of CsrC and CsrB by CRP-cAMP in <i>Salmonella enterica</i>.

Similar Datasets

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