Replication fork blockage by transcription factor-DNA complexes in Escherichia coli

B. T. I. Payne, I. C. Van Knippenberg, H. Bell, S. R. Filipe, D. A. Sherratt, Peter McGlynn

Research output: Contribution to journalArticlepeer-review

39 Citations (Scopus)

Abstract

All organisms require mechanisms that resuscitate replication forks when they break down, reflecting the complex intracellular environments within which DNA replication occurs. Here we show that as few as three lac repressor-operator complexes block Escherichia coli replication forks in vitro regardless of the topological state of the DNA. Blockage with tandem repressor-operator complexes was also observed in vivo, demonstrating that replisomes have a limited ability to translocate through high affinity protein-DNA complexes. However, cells could tolerate tandem repressor-bound operators within the chromosome that were sufficient to block all forks in vitro. This discrepancy between in vitro and in vivo observations was at least partly explained by the ability of RecA, RecBCD and RecG to abrogate the effects of repressor-operator complexes on cell viability. However, neither RuvABC nor RecF were needed for normal cell growth in the face of such complexes. Holliday junction resolution by RuvABC and facilitated loading of RecA by RecF were not therefore critical for tolerance of protein-DNA blocks. We conclude that there is a trade-off between efficient genome duplication and other aspects of DNA metabolism such as transcriptional control, and that recombination enzymes, either directly or indirectly, provide the means to tolerate such conflicts.

Original languageEnglish
Pages (from-to)5194-5202
Number of pages8
JournalNucleic Acids Research
Volume34
Issue number18
DOIs
Publication statusPublished - Oct 2006

Keywords

  • RNA-POLYMERASE
  • RECOMBINATION
  • MODULATION
  • RESTART
  • REPAIR
  • PURIFICATION
  • REQUIREMENT
  • TERMINATION
  • CHROMOSOME
  • PROTEINS

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