Format

Send to

Choose Destination
Elife. 2019 Aug 29;8. pii: e45909. doi: 10.7554/eLife.45909.

Homology sensing via non-linear amplification of sequence-dependent pausing by RecQ helicase.

Author information

1
Laboratory of Single Molecule Biophysics, National Heart, Lung, and Blood Institute, National Institutes of Health, Bethesda, United States.
2
Department of Biochemistry, ELTE-MTA "Momentum" Motor Enzymology Research Group, Eötvös Loránd University, Budapest, Hungary.
3
Department of Biochemistry, MTA-ELTE Motor Pharmacology Research Group, Eötvös Loránd University, Budapest, Hungary.
#
Contributed equally

Abstract

RecQ helicases promote genomic stability through their unique ability to suppress illegitimate recombination and resolve recombination intermediates. These DNA structure-specific activities of RecQ helicases are mediated by the helicase-and-RNAseD like C-terminal (HRDC) domain, via unknown mechanisms. Here, employing single-molecule magnetic tweezers and rapid kinetic approaches we establish that the HRDC domain stabilizes intrinsic, sequence-dependent, pauses of the core helicase (lacking the HRDC) in a DNA geometry-dependent manner. We elucidate the core unwinding mechanism in which the unwinding rate depends on the stability of the duplex DNA leading to transient sequence-dependent pauses. We further demonstrate a non-linear amplification of these transient pauses by the controlled binding of the HRDC domain. The resulting DNA sequence- and geometry-dependent pausing may underlie a homology sensing mechanism that allows rapid disruption of unstable (illegitimate) and stabilization of stable (legitimate) DNA strand invasions, which suggests an intrinsic mechanism of recombination quality control by RecQ helicases.

KEYWORDS:

E. coli; biochemistry; chemical biology; genome stability; helicase; homologous recombination; magnetic tweezers; molecular biophysics; single molecule biophysics; structural biology; unwinding mechanism

PMID:
31464683
PMCID:
PMC6773442
DOI:
10.7554/eLife.45909
[Indexed for MEDLINE]
Free PMC Article

Supplemental Content

Full text links

Icon for eLife Sciences Publications, Ltd Icon for PubMed Central
Loading ...
Support Center