Skip to main content
placeholder image

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

Journal Article


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.

Publication Date


  • 2008

Citation


  • 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.

Scopus Eid


  • 2-s2.0-38849174361

Ro Metadata Url


  • http://ro.uow.edu.au/scipapers/4164

Has Global Citation Frequency


Number Of Pages


  • 6

Start Page


  • 170

End Page


  • 176

Volume


  • 15

Issue


  • 2

Place Of Publication


  • United States

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.

Publication Date


  • 2008

Citation


  • 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.

Scopus Eid


  • 2-s2.0-38849174361

Ro Metadata Url


  • http://ro.uow.edu.au/scipapers/4164

Has Global Citation Frequency


Number Of Pages


  • 6

Start Page


  • 170

End Page


  • 176

Volume


  • 15

Issue


  • 2

Place Of Publication


  • United States