RecG and UvsW catalyse robust DNA rewinding critical for stalled DNA replication fork rescue

Maria Manosas, Senthil K. Perumal, Piero Bianco, Felix Ritort, Stephen J. Benkovic, Vincent Croquette

Research output: Contribution to journalArticlepeer-review

44 Scopus citations

Abstract

Helicases that both unwind and rewind DNA have central roles in DNA repair and genetic recombination. In contrast to unwinding, DNA rewinding by helicases has proved difficult to characterize biochemically because of its thermodynamically downhill nature. Here we use single-molecule assays to mechanically destabilize a DNA molecule and follow, in real time, unwinding and rewinding by two DNA repair helicases, bacteriophage T4 UvsW and Escherichia coli RecG. We find that both enzymes are robust rewinding enzymes, which can work against opposing forces as large as 35 pN, revealing their active character. The generation of work during the rewinding reaction allows them to couple rewinding to DNA unwinding and/or protein displacement reactions central to the rescue of stalled DNA replication forks. The overall results support a general mechanism for monomeric rewinding enzymes.

Original languageEnglish (US)
Article number2368
JournalNature communications
Volume4
DOIs
StatePublished - 2013

All Science Journal Classification (ASJC) codes

  • Chemistry(all)
  • Biochemistry, Genetics and Molecular Biology(all)
  • Physics and Astronomy(all)

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