Molecular analysis of OmpR binding sequences involved in the regulation of ompF in Escherichia coli

Abstract

OmpR, the transcriptional regulatory protein of ompF, had not been previously shown to specifically bind to the −70 to −60-bp region of ompF. We show that the −102 to −76-bp sequence of ompF has a high affinity binding site for OmpR and produced a single OmpR/ ompF complex (complex b). Extension of this DNA fragment to include an inverted repeat sequence located between the −71 and −64-bp region resulted in the formation of a second, slower migrating complex (complex a). A −102 to −58-bp fragment containing a substitution of the −70 CG bp was able to form complex b, but not complex a. A mutant OmpR protein derived from a strain that can not repress ompF was unable to form complex a, while complex b was formed normally. Deletion of the −70 CG bp resulted in incomplete repression of OmpF. These results suggest that OmpR binds to the −71 to −64-bp region and that this sequence plays a role in the regulation of ompF in Escherichia coli.