Identification of the genes in multicopy plasmids affecting ompC and ompF expression in Escherichia coli

Abstract

Osmoregulation of the porin genes, ompF and ompC of Escherichia coli, occurs at the level of transcription through the action of EnvZ and OmpR proteins as well as at the level of translation through micF antisense RNA. In this study, we used a genetic screening approach to identify new genes which interfere with the expression of ompC or ompF. Using an E. coli genomic library in pUC19, we identified three clones whose products altered expression of ompC and ompF in response to medium osmolarity. One clone carrying the secB gene was found to block ompC and inhibit ompF expression. One clone carrying gcvA, a transcriptional regulator for the gvcA operon, was found to block ompF expression at high osmolarity and elevate ompC expression at low osmolarity. One clone carrying rbsR, a repressor for the rbs operon, was found to block ompF expression at both low and high osmolarities and elevate ompC expression at low osmolarity. These results suggest that ompF and ompC expression is associated with other physiological regulating systems in addition to osmoregulation.