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CRP-Mediated Carbon Catabolite Regulation of Yersinia pestis Biofilm Formation Is Enhanced by the Carbon Storage Regulator Protein, CsrA.

Willias SP, Chauhan S, Lo CC, Chain PS, Motin VL - PLoS ONE (2015)

Bottom Line: Media supplementation with cAMP, a small-molecule activator of CRP, did not significantly alter Y. pestis biofilm production.Therefore, our findings indicate CRP does not confer a direct stimulatory effect, but may indirectly promote Y. pestis biofilm production by facilitating the alternate carbon source expression profile.Loss of csrA did not significantly alter relative hmsH, hmsP, or hmsT mRNA abundance.

View Article: PubMed Central - PubMed

Affiliation: Department of Pathology, University of Texas Medical Branch, Galveston, Texas, United States of America.

ABSTRACT
The natural transmission of Yersinia pestis is reliant upon biofilm blockage of the flea vector. However, the environmentally-responsive adaptive regulators which facilitate Y. pestis biofilm production in accordance with the flea midgut milieu are not well understood. We seek to establish the impact of available carbon source metabolism and storage upon Y. pestis biofilm production. Our findings demonstrate that Y. pestis biofilm production is subject to carbon catabolite regulation in which the presence of glucose impairs biofilm production; whereas, the sole metabolism of alternate carbon sources promotes robust biofilm formation. This observation is facilitated by the cAMP receptor protein, CRP. In accordance with a stark growth defect, deletion of crp in both CO92 and KIM6+ Y. pestis strains significantly impaired biofilm production when solely utilizing alternate carbon sources. Media supplementation with cAMP, a small-molecule activator of CRP, did not significantly alter Y. pestis biofilm production. Furthermore, CRP did not alter mRNA abundance of previously-characterized hms biofilm synthesis and regulation factors. Therefore, our findings indicate CRP does not confer a direct stimulatory effect, but may indirectly promote Y. pestis biofilm production by facilitating the alternate carbon source expression profile. Additionally, we assessed the impact of the carbon storage regulator protein, CsrA, upon Y. pestis biofilm production. Contrary to what has been described for E. coli, Y. pestis biofilm formation was found to be enhanced by CsrA. Regardless of media composition and available carbon source, deletion of csrA significantly impaired Y. pestis biofilm production. CsrA was found to promote Y. pestis biofilm production independent of glycogen regulation. Loss of csrA did not significantly alter relative hmsH, hmsP, or hmsT mRNA abundance. However, deletion of hmsP in the csrA-deficient mutant enabled excessive biofilm production, suggesting CsrA enables potent Y. pestis biofilm production through cyclic diguanylate regulation.

No MeSH data available.


Related in: MedlinePlus

Carbon Catabolite Regulation of Y. pestis Biofilm Production.Crystal violet biofilm quantification assay of KIM6+ and CO92 when cultured in BCS medium supplemented with primary (glucose) and/or alternate (glycerol or K-gluconate) carbon sources. Error bars reflect standard deviation from the mean derived from two independent experiments, each consisting of 6 technical replicates. * P-value <0.005 determined by Tukey’s HSD post-hoc analysis.
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pone.0135481.g003: Carbon Catabolite Regulation of Y. pestis Biofilm Production.Crystal violet biofilm quantification assay of KIM6+ and CO92 when cultured in BCS medium supplemented with primary (glucose) and/or alternate (glycerol or K-gluconate) carbon sources. Error bars reflect standard deviation from the mean derived from two independent experiments, each consisting of 6 technical replicates. * P-value <0.005 determined by Tukey’s HSD post-hoc analysis.

Mentions: CO92 and KIM6+ biofilm production was quantified by crystal violet assay during growth in BCS media utilizing either 0.2% glucose or 0.2% alternate carbon sources (Fig 3). After 24 hours post-inoculation, growth of Y. pestis in BCS medium solely supplemented with alternate carbon sources yielded greater than 2-fold enhanced biofilm production relative to the biofilm production in media containing glucose. Moreover, the biofilm production during growth in BCS medium containing a combination of both primary and alternate carbon sources was significantly impaired relative to the biofilm formation when grown in media solely supplemented with alternate carbon sources, indicative of a carbon catabolite repression effect. No change in growth rate was observed for either Y. pestis strain during culture in BCS medium regardless of primary or alternate carbon source supplementation. Therefore, the catabolite regulation of Y. pestis biofilm formation is independent of growth discrepancies.


CRP-Mediated Carbon Catabolite Regulation of Yersinia pestis Biofilm Formation Is Enhanced by the Carbon Storage Regulator Protein, CsrA.

Willias SP, Chauhan S, Lo CC, Chain PS, Motin VL - PLoS ONE (2015)

Carbon Catabolite Regulation of Y. pestis Biofilm Production.Crystal violet biofilm quantification assay of KIM6+ and CO92 when cultured in BCS medium supplemented with primary (glucose) and/or alternate (glycerol or K-gluconate) carbon sources. Error bars reflect standard deviation from the mean derived from two independent experiments, each consisting of 6 technical replicates. * P-value <0.005 determined by Tukey’s HSD post-hoc analysis.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4549057&req=5

pone.0135481.g003: Carbon Catabolite Regulation of Y. pestis Biofilm Production.Crystal violet biofilm quantification assay of KIM6+ and CO92 when cultured in BCS medium supplemented with primary (glucose) and/or alternate (glycerol or K-gluconate) carbon sources. Error bars reflect standard deviation from the mean derived from two independent experiments, each consisting of 6 technical replicates. * P-value <0.005 determined by Tukey’s HSD post-hoc analysis.
Mentions: CO92 and KIM6+ biofilm production was quantified by crystal violet assay during growth in BCS media utilizing either 0.2% glucose or 0.2% alternate carbon sources (Fig 3). After 24 hours post-inoculation, growth of Y. pestis in BCS medium solely supplemented with alternate carbon sources yielded greater than 2-fold enhanced biofilm production relative to the biofilm production in media containing glucose. Moreover, the biofilm production during growth in BCS medium containing a combination of both primary and alternate carbon sources was significantly impaired relative to the biofilm formation when grown in media solely supplemented with alternate carbon sources, indicative of a carbon catabolite repression effect. No change in growth rate was observed for either Y. pestis strain during culture in BCS medium regardless of primary or alternate carbon source supplementation. Therefore, the catabolite regulation of Y. pestis biofilm formation is independent of growth discrepancies.

Bottom Line: Media supplementation with cAMP, a small-molecule activator of CRP, did not significantly alter Y. pestis biofilm production.Therefore, our findings indicate CRP does not confer a direct stimulatory effect, but may indirectly promote Y. pestis biofilm production by facilitating the alternate carbon source expression profile.Loss of csrA did not significantly alter relative hmsH, hmsP, or hmsT mRNA abundance.

View Article: PubMed Central - PubMed

Affiliation: Department of Pathology, University of Texas Medical Branch, Galveston, Texas, United States of America.

ABSTRACT
The natural transmission of Yersinia pestis is reliant upon biofilm blockage of the flea vector. However, the environmentally-responsive adaptive regulators which facilitate Y. pestis biofilm production in accordance with the flea midgut milieu are not well understood. We seek to establish the impact of available carbon source metabolism and storage upon Y. pestis biofilm production. Our findings demonstrate that Y. pestis biofilm production is subject to carbon catabolite regulation in which the presence of glucose impairs biofilm production; whereas, the sole metabolism of alternate carbon sources promotes robust biofilm formation. This observation is facilitated by the cAMP receptor protein, CRP. In accordance with a stark growth defect, deletion of crp in both CO92 and KIM6+ Y. pestis strains significantly impaired biofilm production when solely utilizing alternate carbon sources. Media supplementation with cAMP, a small-molecule activator of CRP, did not significantly alter Y. pestis biofilm production. Furthermore, CRP did not alter mRNA abundance of previously-characterized hms biofilm synthesis and regulation factors. Therefore, our findings indicate CRP does not confer a direct stimulatory effect, but may indirectly promote Y. pestis biofilm production by facilitating the alternate carbon source expression profile. Additionally, we assessed the impact of the carbon storage regulator protein, CsrA, upon Y. pestis biofilm production. Contrary to what has been described for E. coli, Y. pestis biofilm formation was found to be enhanced by CsrA. Regardless of media composition and available carbon source, deletion of csrA significantly impaired Y. pestis biofilm production. CsrA was found to promote Y. pestis biofilm production independent of glycogen regulation. Loss of csrA did not significantly alter relative hmsH, hmsP, or hmsT mRNA abundance. However, deletion of hmsP in the csrA-deficient mutant enabled excessive biofilm production, suggesting CsrA enables potent Y. pestis biofilm production through cyclic diguanylate regulation.

No MeSH data available.


Related in: MedlinePlus