Title

Single-molecule studies of fork dynamics in Escherichia coli DNA replication

RIS ID

22378

Publication Details

Tanner, N. A., Hamdan, S. M., Jergic, S., Schaeffer, P. M., Dixon, N. E. & van Oijen, A. M. (2008). Single-molecule studies of fork dynamics in Escherichia coli DNA replication. Nature Structural and Molecular Biology, 15 (2), 170-176.

Abstract

We present single-molecule studies of the Escherichia coli replication machinery. We visualize individual E. coli DNA polymerase III (Pol III) holoenzymes engaging in primer extension and leading-strand synthesis. When coupled to the replicative helicase DnaB, Pol III mediates leading-strand synthesis with a processivity of 10.5 kilobases (kb), eight-fold higher than that by Pol III alone. Addition of the primase DnaG causes a three-fold reduction in the processivity of leading-strand synthesis, an effect dependent upon the DnaB-DnaG protein-protein interaction rather than primase activity. A single-molecule analysis of the replication kinetics with varying DnaG concentrations indicates that a cooperative binding of two or three DnaG monomers to DnaB halts synthesis. Modulation of DnaB helicase activity through the interaction with DnaG suggests a mechanism that prevents leading-strand synthesis from outpacing lagging-strand synthesis during slow primer synthesis on the lagging strand.

Grant Number

ARC/LX0882706

Please refer to publisher version or contact your library.

Share

COinS