Single-molecule analysis reveals that the lagging strand increases replisome processivity but slows replication fork progression

Proc Natl Acad Sci U S A. 2009 Aug 11;106(32):13236-41. doi: 10.1073/pnas.0906157106. Epub 2009 Aug 3.

Abstract

Single-molecule techniques are developed to examine mechanistic features of individual E. coli replisomes during synthesis of long DNA molecules. We find that single replisomes exhibit constant rates of fork movement, but the rates of different replisomes vary over a surprisingly wide range. Interestingly, lagging strand synthesis decreases the rate of the leading strand, suggesting that lagging strand operations exert a drag on replication fork progression. The opposite is true for processivity. The lagging strand significantly increases the processivity of the replisome, possibly reflecting the increased grip to DNA provided by 2 DNA polymerases anchored to sliding clamps on both the leading and lagging strands.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • DNA Polymerase III / metabolism
  • DNA Replication*
  • DNA, Circular / metabolism
  • DNA-Directed DNA Polymerase / metabolism*
  • Diffusion
  • DnaB Helicases / metabolism
  • Escherichia coli / enzymology
  • Escherichia coli / metabolism*
  • Lipid Bilayers / metabolism
  • Multienzyme Complexes / metabolism*

Substances

  • DNA, Circular
  • Lipid Bilayers
  • Multienzyme Complexes
  • DNA synthesome
  • DNA Polymerase III
  • DNA-Directed DNA Polymerase
  • dnaB protein, E coli
  • DnaB Helicases