BRCA2 function in DNA binding and recombination from a BRCA2-DSS1-ssDNA structure

Haijuan Yang; Philip D Jeffrey; Julie Miller; Elspeth Kinnucan; Yutong Sun; Nicolas H Thoma; Ning Zheng; Phang-Lang Chen; Wen-Hwa Lee; Nikola P Pavletich

doi:10.1126/science.297.5588.1837

BRCA2 function in DNA binding and recombination from a BRCA2-DSS1-ssDNA structure

Science. 2002 Sep 13;297(5588):1837-48. doi: 10.1126/science.297.5588.1837.

Authors

Haijuan Yang¹, Philip D Jeffrey, Julie Miller, Elspeth Kinnucan, Yutong Sun, Nicolas H Thoma, Ning Zheng, Phang-Lang Chen, Wen-Hwa Lee, Nikola P Pavletich

Affiliation

¹ Department of Pharmacology, Sloan-Kettering Division, Joan and Sanford I. Weill Graduate School of Medical Sciences, Cornell University, New York, NY 10021, USA.

PMID: 12228710
DOI: 10.1126/science.297.5588.1837

Abstract

Mutations in the BRCA2 (breast cancer susceptibility gene 2) tumor suppressor lead to chromosomal instability due to defects in the repair of double-strand DNA breaks (DSBs) by homologous recombination, but BRCA2's role in this process has been unclear. Here, we present the 3.1 angstrom crystal structure of a approximately 90-kilodalton BRCA2 domain bound to DSS1, which reveals three oligonucleotide-binding (OB) folds and a helix-turn-helix (HTH) motif. We also (i) demonstrate that this BRCA2 domain binds single-stranded DNA, (ii) present its 3.5 angstrom structure bound to oligo(dT)9, (iii) provide data that implicate the HTH motif in dsDNA binding, and (iv) show that BRCA2 stimulates RAD51-mediated recombination in vitro. These findings establish that BRCA2 functions directly in homologous recombination and provide a structural and biochemical basis for understanding the loss of recombination-mediated DSB repair in BRCA2-associated cancers.

Publication types

Research Support, Non-U.S. Gov't
Research Support, U.S. Gov't, P.H.S.

MeSH terms

Amino Acid Sequence
Animals
BRCA2 Protein / chemistry*
BRCA2 Protein / genetics
BRCA2 Protein / metabolism*
Binding Sites
Cell Cycle Proteins
Crystallography, X-Ray
DNA / metabolism
DNA Repair*
DNA, Single-Stranded / metabolism*
DNA-Binding Proteins / metabolism
Genes, BRCA2
Helix-Turn-Helix Motifs
Humans
Hydrogen Bonding
Hydrophobic and Hydrophilic Interactions
Mice
Molecular Sequence Data
Mutation
Proteasome Endopeptidase Complex
Protein Conformation
Protein Folding
Protein Structure, Secondary
Protein Structure, Tertiary
Proteins / chemistry
Proteins / metabolism*
Rad51 Recombinase
Rats
Recombination, Genetic*

Substances

BRCA2 Protein
Cell Cycle Proteins
DNA, Single-Stranded
DNA-Binding Proteins
Proteins
SEM1 protein, human
Shfdg1 protein, mouse
DNA
RAD51 protein, human
Rad51 Recombinase
Rad51 protein, mouse
Rad51 protein, rat
Proteasome Endopeptidase Complex

Associated data

PDB/1IYJ
PDB/1MIU
PDB/1MJE