Repressor CopG prevents access of RNA polymerase to promoter and actively dissociates open complexes

Abstract

Replication of the promiscuous plasmid pMV158 requires expression of the initiator repB gene, which is controlled by the repressor CopG. Genes repB and copG are co-transcribed from promoter Pcr. We have studied the interactions between RNA polymerase, CopG and the promoter to elucidate the mechanism of repression by CopG. Complexes formed at 0°C and at 37°C between RNA polymerase and Pcr differed from each other in stability and in the extent of the DNA contacted. The 37°C complex was very stable (half-life of about 3 h), and shared features with typical open complexes generated at a variety of promoters. CopG protein repressed transcription from Pcr at two different stages in the process leading to the initiation complex. First, CopG hindered binding of RNA polymerase to the promoter. Second, CopG was able to displace RNA polymerase once the enzyme has formed a stable complex with Pcr. A model for the CopG-mediated disassembly of the stable RNA polymerase–Pcr promoter complex is presented.