Format

Send to

Choose Destination
Cell. 2015 Jul 16;162(2):271-286. doi: 10.1016/j.cell.2015.06.015.

Rad51 Paralogs Remodel Pre-synaptic Rad51 Filaments to Stimulate Homologous Recombination.

Author information

1
DNA Damage Response Laboratory, Clare Hall Laboratory, The Francis Crick Institute, South Mimms EN6 3LD, UK.
2
Department of Biology, Masaryk University, 62500 Brno, Czech Republic; International Clinical Research Center, St. Anne's University Hospital in Brno, 62500 Brno, Czech Republic.
3
Section of Virology, Single Molecule Imaging Group and MRC Clinical Sciences Centre, Department of Medicine, Imperial College London, London W12 0NN, UK.
4
DNA Damage Response Laboratory, Clare Hall Laboratory, The Francis Crick Institute, South Mimms EN6 3LD, UK; UCSF-Mission Bay, Genentech Hall S574, San Francisco, CA 94158, USA.
5
Electron Microscopy Science Technology Platform, Lincoln's Inn Fields Laboratory, The Francis Crick Institute, London WC2A 3LY, UK.
6
Department of Biochemistry and Molecular Genetics, University of Virginia School of Medicine, Charlottesville, VA 22908, USA.
7
Department of Biology, Masaryk University, 62500 Brno, Czech Republic; International Clinical Research Center, St. Anne's University Hospital in Brno, 62500 Brno, Czech Republic; National Centre for Biomolecular Research, Masaryk University, 62500 Brno, Czech Republic. Electronic address: lkrejci@chemi.muni.cz.
8
DNA Damage Response Laboratory, Clare Hall Laboratory, The Francis Crick Institute, South Mimms EN6 3LD, UK. Electronic address: simon.boulton@crick.ac.uk.

Abstract

Repair of DNA double strand breaks by homologous recombination (HR) is initiated by Rad51 filament nucleation on single-stranded DNA (ssDNA), which catalyzes strand exchange with homologous duplex DNA. BRCA2 and the Rad51 paralogs are tumor suppressors and critical mediators of Rad51. To gain insight into Rad51 paralog function, we investigated a heterodimeric Rad51 paralog complex, RFS-1/RIP-1, and uncovered the molecular basis by which Rad51 paralogs promote HR. Unlike BRCA2, which nucleates RAD-51-ssDNA filaments, RFS-1/RIP-1 binds and remodels pre-synaptic filaments to a stabilized, "open," and flexible conformation, in which the ssDNA is more accessible to nuclease digestion and RAD-51 dissociation rate is reduced. Walker box mutations in RFS-1, which abolish filament remodeling, fail to stimulate RAD-51 strand exchange activity, demonstrating that remodeling is essential for RFS-1/RIP-1 function. We propose that Rad51 paralogs stimulate HR by remodeling the Rad51 filament, priming it for strand exchange with the template duplex.

PMID:
26186187
PMCID:
PMC4518479
DOI:
10.1016/j.cell.2015.06.015
[Indexed for MEDLINE]
Free PMC Article

Supplemental Content

Full text links

Icon for Elsevier Science Icon for PubMed Central
Loading ...
Support Center