skip to main content
OSTI.GOV title logo U.S. Department of Energy
Office of Scientific and Technical Information

Title: The BRCA2 homologue Brh2 nucleates RAD51 filament formation at a dsDNA–ssDNA junction

Journal Article · · Nature (London)
 [1];  [1];  [1];  [2];  [3]
  1. Memorial Sloan-Kettering Cancer Center, New York, NY (United States). Structural Biology Program
  2. Cornell University Weill Medical College, New York, NY (United States). Hearst Microbiology Research Center
  3. Memorial Sloan-Kettering Cancer Center, New York, NY (United States). Structural Biology Program; Memorial Sloan-Kettering Cancer Center, NY, NY (United States). Howard Hughes Medical Institute
+ Show Author Affiliations

The BRCA2 tumour suppressor is known to be essential for the error-free repair of double-strand breaks (DSBs) in DNA by homologous recombination. This is mediated by RAD51, which forms a nucleoprotein filament with the 3' overhanging single-stranded DNA (ssDNA) of the resected DSB, searches for a homologous donor sequence, and catalyses strand exchange with the donor DNA. The 3,418-amino-acid BRCA2 contains eight ~30-amino-acid BRC repeats that bind RAD51 and a ~700-amino-acid DBD domain that binds ssDNA7. The isolated BRC and DBD domains have the opposing effects of inhibiting and stimulating recombination, respectively, and the role of BRCA2 in repair has been unclear. Here we show that a full-length BRCA2 homologue (Brh2) stimulates Rad51-mediated recombination at substoichiometric concentrations relative to Rad51. Brh2 recruits Rad51 to DNA and facilitates the nucleation of the filament, which is then elongated by the pool of free Rad51. Brh2 acts preferentially at a junction between double-stranded DNA (dsDNA) and ssDNA, with strict specificity for the 3' overhang polarity of a resected DSB. These results establish a BRCA2 function in RAD51-mediated DSB repair and explain the loss of this repair capacity in BRCA2-associated cancers.

Research Organization:
Brookhaven National Lab. (BNL), Upton, NY (United States). National Synchrotron Light Source (NSLS)
Sponsoring Organization:
USDOE Office of Science (SC)
DOE Contract Number:
AC02-98CH10886
OSTI ID:
884535
Report Number(s):
BNL-76663-2005-JA
Journal Information:
Nature (London), Vol. 433, Issue 7026; ISSN 0028-0836
Publisher:
Nature Publishing Group
Country of Publication:
United States
Language:
English

References (27)

The Genetics of Breast Cancer Susceptibility journal December 1998
Mutation in Brca2 stimulates error-prone homology-directed repair of DNA double-strand breaks occurring between repeated sequences journal September 2001
Rad52 Protein Associates with Replication Protein A (RPA)-Single-stranded DNA to Accelerate Rad51-mediated Displacement of RPA and Presynaptic Complex Formation journal June 2002
Targeted Inactivation of Mouse RAD52 Reduces Homologous Recombination but Not Resistance to Ionizing Radiation journal November 1998
A Single-stranded DNA-binding Protein Is Needed for Efficient Presynaptic Complex Formation by the Saccharomyces cerevisiae Rad51 Protein journal March 1997
Sequence fingerprints in BRCA2 and RAD51: implications for DNA repair and cancer journal September 2003
Function of Yeast Rad52 Protein as a Mediator between Replication Protein A and the Rad51 Recombinase journal November 1997
Rad52 protein stimulates DNA strand exchange by Rad51 and replication protein A journal January 1998
Molecular views of recombination proteins and their control journal June 2003
BRCA2 Homolog Required for Proficiency in DNA Repair, Recombination, and Genome Stability in Ustilago maydis journal September 2002
Insights into DNA recombination from the structure of a RAD51–BRCA2 complex journal November 2002
The BRC repeats in BRCA2 are critical for RAD51 binding and resistance to methyl methanesulfonate treatment journal April 1998
In Vivo Roles of Rad52, Rad54, and Rad55 Proteins in Rad51-Mediated Recombination journal July 2003
The BRCA2-Interacting Protein DSS1 Is Vital for DNA Repair, Recombination, and Genome Stability in Ustilago maydis journal October 2003
Rapid Exchange of A:T Base Pairs Is Essential for Recognition of DNA Homology by Human Rad51 Recombination Protein journal November 1999
RAD51 Interacts with the Evolutionarily Conserved BRC Motifs in the Human Breast Cancer Susceptibility Gene brca2 journal December 1997
Reconstitution of the strand invasion step of double-strand break repair using human Rad51 Rad52 and RPA proteins journal November 2000
Choreography of the DNA Damage Response journal September 2004
RecFOR Proteins Load RecA Protein onto Gapped DNA to Accelerate DNA Strand Exchange journal May 2003
Role of BRCA2 in Control of the RAD51 Recombination and DNA Repair Protein journal February 2001
BRCA2 Is Required for Homology-Directed Repair of Chromosomal Breaks journal February 2001
BRCA2 Function in DNA Binding and Recombination from a BRCA2-DSS1-ssDNA Structure journal September 2002
Expression of BRC Repeats in Breast Cancer Cells Disrupts the BRCA2-Rad51 Complex and Leads to Radiation Hypersensitivity and Loss of G 2 /M Checkpoint Control journal November 1999
Recruitment of the Recombinational Repair Machinery to a DNA Double-Strand Break in Yeast journal July 2003
Stimulation by Rad52 of yeast Rad51- mediated recombination journal January 1998
DNA strand exchange mediated by a RAD51-ssDNA nucleoprotein filament with polarity opposite to that of RecA journal August 1995
Role of Saccharomyces Single-Stranded DNA-Binding Protein RPA in the Strand Invasion Step of Double-Strand Break Repair journal January 2004

Similar Records

Identification of Rad51 regulation by BRCA2 using Caenorhabditis elegans BRCA2 and bimolecular fluorescence complementation analysis
Journal Article · Sat Nov 03 00:00:00 EDT 2007 · Biochemical and Biophysical Research Communications · OSTI ID:884535
+2 more
Mechanism of homologous recombination from the RecA-ssDNA/dsDNA structures
Journal Article · Tue Jul 08 00:00:00 EDT 2008 · Nature · OSTI ID:884535
Promotion of Homologous Recombination and Genomic Stability byRAD51AP1 via RAD51 Recombinase Enhancement
Journal Article · Wed Apr 11 00:00:00 EDT 2007 · Molecular Cell · OSTI ID:884535
+8 more

Related Subjects

59 BASIC BIOLOGICAL SCIENCES
DNA
FILAMENTS
BIOSYNTHESIS
national synchrotron light source