In vivo relations between pAMβ1‐encoded type I topoisomerase and plasmid replication

Abstract

A number of large extrachromosomal elements encode prokaryotic type I topoisomerases of unknown functions. Here, we analysed the topoisomerase Topβ encoded by the Gram-positive broad-host-range plasmid pAMβ1. We show that this enzyme possesses the DNA relaxation activity of type I topoisomerases. Interestingly, it is active only on plasmids that use DNA polymerase I to initiate replication, such as pAMβ1, and depends on the activity of this polymerase. This is the first example, to our knowledge, of prokaryotic type I topoisomerase that is specific for a given type of replicon. During pAMβ1 replication in Bacillus subtilis cells, Topβ promotes premature arrest of DNA polymerase I, ≈190 bp downstream of the replication initiation point. We propose that Topβ acts on the early replication intermediates of pAMβ1, which contain D-loops formed by DNA polymerase I-mediated strand displacement. The possible role of the resulting DNA Pol I arrest in plasmid replication is discussed.