Interplay between Type 1A Topoisomerases and Gyrase in Chromosome Segregation in Escherichia coli

Abstract

ABSTRACT Escherichia coli possesses two type 1A topoisomerases, Topo I ( topA ) and Topo III ( topB ). Topo I relaxes excess negative supercoiling, and topA mutants can grow only in the presence of compensatory mechanisms, such as gyrase mutations. topB mutants grow as well as wild-type cells. In vitro , Topo III, but not Topo I, can efficiently decatenate DNA during replication. However, in vivo , a chromosome segregation defect is seen only when both type 1A topoisomerases are absent. Here we present experimental evidence for an interplay between gyrase and type 1A topoisomerases in chromosome segregation. We found that both the growth defect and the Par − phenotypes of a gyrB (Ts) mutant at nonpermissive temperatures were significantly corrected by deleting topA , but only when topB was present. Overproducing Topo IV, the major cellular decatenase, could not substitute for topB . We also show that overproducing Topo III at a very high level could suppress the Par − phenotype. We previously found that the growth and chromosome segregation defects of a triple topA rnhA gyrB (Ts) mutant in which gyrase supercoiling activity was strongly inhibited could be corrected by overproducing Topo III (V. Usongo, F. Nolent, P. Sanscartier, C. Tanguay, S. Broccoli, I. Baaklini, K. Drlica, and M. Drolet, Mol. Microbiol. 69: 968-981, 2008). We show here that this overproduction could be bypassed by substituting the gyrB (Ts) allele for a gyrB + one or by growing cells in a minimal medium, conditions that reduced both topA - and rnhA -dependent unregulated replication. Altogether, our data point to a role for Topo III in chromosome segregation when gyrase is inefficient and suggest that Topo I plays an indirect role via supercoiling regulation.