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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.

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