RecG and UvsW Catalyse Robust DNA Rewinding Critical for Stalled DNA Replication Fork Rescue

Overview

Journal Nat Commun

Specialty Biology

Date 2013 Sep 10

PMID 24013402

Citations 50

Authors

Maria Manosas

Senthil K Perumal

Piero R Bianco

Piero Bianco

Felix Ritort

Stephen J Benkovic

Vincent Croquette

Affiliations

Soon will be listed here.

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.

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