The Relative Contributions of Transcription and Translation to Plasmid DNA Supercoiling in Salmonella Typhimurium

Mutations affecting DNA topoisomerase I (topA) in Salmonella typhimurium were isolated and graded on the basis of their ability to reverse the effects of gyrB mutations on his operon expression. Different topA and gyrB alleles (in otherwise isogenic strains) were used to gather insights into the transcription-dependent variability of plasmid DNA-linking deficit in growing bacteria. This study showed that modulation of DNA supercoiling by transcription results from the action of two components: one is highly dependent on the coupling of translation to RNA-chain elongation; and the other is unrelated to protein synthesis and entirely dependent on promoter determinants. The former greatly predominates in DNA topoisomerase I mutants (topA and topA gyrB) while the latter is the sole contributor to plasmid DNA-linking deficit in wild-type cells. Altogether, these data suggest that whereas translation acts by enhancing the formation of twin supercoiled domains during elongation, the promoter-dependent effects bear no relation to the twin-supercoiled-domain model and are better explained by a mechanism which responds to the binding/unwinding of template DNA by RNA polymerase.