Strand switching of a replicative DNA helicase promoted by the E. coli primosome.

Abstract

The E. coli primosome assembles at an origin on a single-stranded DNA, like that of phi X174, to promote replication of that template. Upon conversion to the duplex form, the primosome can generate a rolling circle product from this template. Rolling circle synthesis implies the transfer of the DnaB helicase from its initial loading site on the viral strand to a displaced complementary strand. Isolated primosomes promote only unit-length synthesis; supplementation with PriC, DnaC, and DnaT is necessary to reconstitute rolling circle synthesis. Rolling circle replication is sensitive to salts, whereas primosome assembly and unit-length synthesis are not. Thus, the primosome promotes two distinct reactions: assembly for first-round synthesis and strand switching for rolling circle synthesis.