rifampin and 2-nitrophenyl-beta-fucoside

Genes are often transcribed by multiple RNA polymerases (RNAPs) at densities that can vary widely across genes and environmental conditions. Here, we provide in vitro and in vivo evidence for a built-in mechanism by which co-transcribing RNAPs display either collaborative or antagonistic dynamics over long distances (>2 kb) through transcription-induced DNA supercoiling. In Escherichia coli, when the promoter is active, co-transcribing RNAPs translocate faster than a single RNAP, but their average speed is not altered by large variations in promoter strength and thus RNAP density. Environmentally induced promoter repression reduces the elongation efficiency of already-loaded RNAPs, causing premature termination and quick synthesis arrest of no-longer-needed proteins. This negative effect appears independent of RNAP convoy formation and is abrogated by topoisomerase I activity. Antagonistic dynamics can also occur between RNAPs from divergently transcribed gene pairs. Our findings may be broadly applicable given that transcription on topologically constrained DNA is the norm across organisms.

Topics: DNA-Directed RNA Polymerases; DNA, Bacterial; DNA, Superhelical; Escherichia coli; Gene Expression Regulation, Bacterial; Glucose; Glycosides; Isopropyl Thiogalactoside; Kinetics; Lac Operon; Plasmids; Promoter Regions, Genetic; Real-Time Polymerase Chain Reaction; Rifampin; RNA, Bacterial; Transcription, Genetic