| Summary: | <p>Abstract</p> <p>Background</p> <p>In <it>Enterococcus faecalis </it>the genes encoding the enzymes involved in citrate metabolism are organized in two divergent operons, <it>citHO </it>and <it>oadHDB-citCDEFX-oadA-citMG </it>(<it>citCL </it>locus). Expression of both operons is specifically activated by adding citrate to the medium. This activation is mediated by binding of the GntR-like transcriptional regulator (CitO) to the <it>cis</it>-acting sequences located in the <it>cit </it>intergenic region. Early studies indicated that citrate and glucose could not be co-metabolized suggesting some form of catabolite repression, however the molecular mechanism remained unknown.</p> <p>Results</p> <p>In this study, we observed that the <it>citHO </it>promoter is repressed in the presence of sugars transported by the Phosphoenolpyruvate:carbohydrate Phosphotranserase System (PTS sugars). This result strongly suggested that Carbon Catabolic Repression (CCR) impedes the expression of the activator CitO and the subsequent induction of the <it>cit </it>pathway. In fact, we demonstrate that CCR is acting on both promoters. It is partially relieved in a <it>ccpA</it>-deficient <it>E. faecalis </it>strain indicating that a CcpA-independent mechanism is also involved in regulation of the two operons. Furthermore, sequence analysis of the <it>citH</it>/<it>oadH </it>intergenic region revealed the presence of three putative catabolite responsive elements (<it>cre</it>). We found that they are all active and able to bind the CcpA/P-Ser-HPr complex, which downregulates the expression of the <it>cit </it>operons. Systematic mutation of the CcpA/P-Ser-HPr binding sites revealed that <it>cre1 </it>and <it>cre2 </it>contribute to <it>citHO </it>repression, while <it>cre3 </it>is involved in CCR of <it>citCL</it></p> <p>Conclusion</p> <p>In conclusion, our study establishes that expression of the <it>cit </it>operons in <it>E. faecalis </it>is controlled by CCR via CcpA-dependent and -independent mechanisms.</p>
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