Replisome speed determines the efficiency of the Tus-Ter replication termination barrier

Nature. 2015 Sep 17;525(7569):394-8. doi: 10.1038/nature14866. Epub 2015 Aug 31.

Abstract

In all domains of life, DNA synthesis occurs bidirectionally from replication origins. Despite variable rates of replication fork progression, fork convergence often occurs at specific sites. Escherichia coli sets a 'replication fork trap' that allows the first arriving fork to enter but not to leave the terminus region. The trap is set by oppositely oriented Tus-bound Ter sites that block forks on approach from only one direction. However, the efficiency of fork blockage by Tus-Ter does not exceed 50% in vivo despite its apparent ability to almost permanently arrest replication forks in vitro. Here we use data from single-molecule DNA replication assays and structural studies to show that both polarity and fork-arrest efficiency are determined by a competition between rates of Tus displacement and rearrangement of Tus-Ter interactions that leads to blockage of slower moving replisomes by two distinct mechanisms. To our knowledge this is the first example where intrinsic differences in rates of individual replisomes have different biological outcomes.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Base Sequence
  • Binding, Competitive
  • Chromosomes, Bacterial / genetics
  • Chromosomes, Bacterial / metabolism
  • Crystallography, X-Ray
  • DNA Replication*
  • DNA-Directed DNA Polymerase / chemistry
  • DNA-Directed DNA Polymerase / metabolism*
  • Escherichia coli / genetics*
  • Escherichia coli / metabolism
  • Escherichia coli Proteins / chemistry
  • Escherichia coli Proteins / metabolism*
  • Kinetics
  • Models, Biological
  • Models, Molecular
  • Movement
  • Multienzyme Complexes / chemistry
  • Multienzyme Complexes / metabolism*
  • Protein Conformation
  • Regulatory Sequences, Nucleic Acid / genetics*
  • Surface Plasmon Resonance
  • Time Factors

Substances

  • Escherichia coli Proteins
  • Multienzyme Complexes
  • tus protein, E coli
  • DNA synthesome
  • DNA-Directed DNA Polymerase

Associated data

  • PDB/4XR0
  • PDB/4XR1
  • PDB/4XR2
  • PDB/4XR3