In vitro binding of the CcpA protein ofBacillus megateriumtocis-acting catabolite responsive elements (CREs) of Gram-positive bacteria

Abstract
Using DNA band migration retardation assays, specific binding of the CcpA protein of Bacillus megaterium to the ds-acting catabolite responsive element (CRE) of the xyl operon of B. subtilis has been demonstrated. Binding of CcpA was specifically inhibited by addition of unlabeled DNA fragments containing CREs of other operons but not by DNA fragments lacking a CRE. Binding was stimulated by high concentrations of phosphate, pyrophosphate, and organic phosphate esters and specifically inhibited by serine phosphorylated HPr and its conformational analogue, S46D HPr. This report therefore documents the specific binding of CcpA to a target CRE and defines its regulation by HPr(ser-P) and phosphorylated metabolites.

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