Effects of the P1 Plasmid Centromere on Expression of P1 Partition Genes

Abstract

ABSTRACT The partition operon of P1 plasmid encodes two proteins, ParA and ParB, required for the faithful segregation of plasmid copies to daughter cells. The operon is followed by a centromere analog, parS , at which ParB binds. ParA, a weak ATPase, represses the par promoter most effectively in its ADP-bound form. ParB can recruit ParA to parS , stimulate its ATPase, and significantly stimulate the repression. We report here that parS also participates in the regulation of expression of the par genes. A single chromosomal parS was shown to augment repression of several copies of the par promoter by severalfold. The repression increase was sensitive to the levels of ParA and ParB and to their ratio. The increase may be attributable to a conformational change in ParA mediated by the parS -ParB complex, possibly acting catalytically. We also observed an in cis effect of parS which enhanced expression of parB , presumably due to a selective modulation of the mRNA level. Although ParB had been earlier found to spread into and silence genes flanking parS , silencing of the par operon by ParB spreading was not significant. Based upon analogies between partitioning and septum placement, we speculate that the regulatory switch controlled by the parS -ParB complex might be essential for partitioning itself.