title: tau couples the leading- and lagging-strand polymerases at the Escherichia coli DNA replication fork.
abstract: Synthesis of an Okazaki fragment occurs once every 1 or 2 s at the Escherichia coli replication fork. To account for the rapid recycling required of the lagging-strand polymerase, it has been proposed that it is held at the replication fork by protein-protein interactions with the leading-strand polymerase as part of a dimeric polymerase assembly. Solution studies showed that the replicative polymerase, the DNA polymerase III holoenzyme, was indeed a dimer with two catalytic cores held together by the tau subunit. However, the functionality of this arrangement at the replication fork has never been demonstrated. We showed previously that the lagging-strand polymerase acted processively during multiple rounds of Okazaki fragment synthesis, i.e. the same polymerase core assembly synthesized each and every fragment made by the fork. Using extreme dilution of active replication forks and the isolation of protein-DNA complexes capable of supporting coupled leading- and lagging-strand synthesis, we demonstrate here that this coupling of leading- and lagging-strand synthesis is, in fact, mediated by the tau subunit of the holoenzyme acting as a physical bridge between the core assemblies synthesizing the leading and lagging strands.
authors.affiliation: Graduate Program in Molecular Biology Cornell University Graduate School of Medical Sciences, New York, New York 10021, USA.
authors.forename: S
authors.initials: S
authors.lastname: Kim
authors.forename: H G
authors.initials: HG
authors.lastname: Dallmann
authors.forename: C S
authors.initials: CS
authors.lastname: McHenry
authors.forename: K J
authors.initials: KJ
authors.lastname: Marians
mesh_terms: D004247:DNA / Q000378:metabolism; D004258:DNA Polymerase III / Q000378:metabolism*; D004261:DNA Replication; D004269:DNA, Bacterial; D004926:Escherichia coli / Q000235:genetics*; D011485:Protein Binding
chemical_list: D004269:DNA, Bacterial; C022262:Okazaki fragments; D004247:DNA; D004258:DNA Polymerase III