Purified human BRCA2 stimulates RAD51-mediated recombination
Open accessAbstract
Mutation of the breast cancer susceptibility gene, BRCA2, leads to breast and ovarian cancers. Mechanistic insight into the functions of human BRCA2 has been limited by the difficulty of isolating this large protein (3,418 amino acids). Here we report the purification of full-length BRCA2 and show that it both binds RAD51 and potentiates recombinational DNA repair by promoting assembly of RAD51 onto single-stranded DNA (ssDNA). BRCA2 acts by targeting RAD51 to ssDNA over double-stranded DNA, enabling RAD51 to displace replication protein-A (RPA) from ssDNA and stabilizing RAD51-ssDNA filaments by blocking ATP hydrolysis. BRCA2 does not anneal ssDNA complexed with RPA, implying it does not directly function in repair processes that involve ssDNA annealing. Our findings show that BRCA2 is a key mediator of homologous recombination, and they provide a molecular basis for understanding how this DNA repair process is disrupted by BRCA2 mutations, which lead to chromosomal instability and cancer.
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Unlike BRCA2-DSS1, BRCA1-BARD1 fails to promote assembly of RAD51 presynaptic filaments on RPA-coated ssDNA.
“while, as reported previously36,38, the BRCA2-DSS1 complex promotes RAD51 presynaptic filament assembly on RPA-coated ssDNA, BRCA1-BARD1 does not possess such an attribute”
From BRCA1–BARD1 promotes RAD51-mediated homologous DNA pairing (2017) · cites this paper as [38]
When dsDNA is present, BRCA1-BARD1 cannot target RAD51 to ssDNA, whereas BRCA2-DSS1 can.
“unlike BRCA2-DSS137,38, BRCA1-BARD1 is devoid of the ability to target RAD51 to ssDNA when dsDNA is present”
From BRCA1–BARD1 promotes RAD51-mediated homologous DNA pairing (2017) · cites this paper as [38]
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- Authors
- Jensen, R. B., Carreira, A., & Kowalczykowski, S. C.
- Affiliations
- University of California, Davis (US)
- Corresponding author
- Kowalczykowski, Stephen C.
- Venue
- Nature
- Published
- 2010-08-22
- Volume
- 467
- Issue
- 7316
- Pages
- 678-683
- ISSN
- 0028-0836 (ISSN-L)
- 1476-4687
- PMC
- PMC2952063
- OpenAlex
- W2085544302
- Cited by
- 715
- References
- 52
- Citations by year
- 2026: 30 · 2025: 32 · 2024: 45 · 2023: 44 · 2022: 43 · 2021: 58 · 2020: 61 · 2019: 41 · 2018: 39 · 2017: 51 · 2016: 54 · 2015: 39 · 2014: 39 · 2013: 43 · 2012: 34
- FWCI
- 18.10
- Type
- journal article
- Language
- en
- Open access
- green
- www.ncbi.nlm.nih.gov
- Licence
- www.springer.com/tdm (tdm)
- Locations
- Nature (publishedVersion)
- PubMed (publishedVersion)
- Europe PMC (PubMed Central) (acceptedVersion)
- PubMed Central (acceptedVersion)
- RePEc: Research Papers in Economics (submittedVersion)
- eScholarship (California Digital Library) (submittedVersion)
- Funders
- National Institute of General Medical Sciences (GM-62653, R37GM062653, R01GM041347, R01GM062653)
- Topics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › DNA Repair Mechanisms
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Genetics › BRCA gene mutations in cancer
- Health Sciences › Medicine › Oncology › PARP inhibition in cancer therapy
- Keywords
- BRCA2, RAD51, homologous recombination, recombinational DNA repair, single-stranded DNA, replication protein A, ATP hydrolysis, genomic instability, breast cancer susceptibility, ovarian cancer, protein purification
- MeSH terms
- Cell Line, DNA, DNA Repair, DNA-Binding Proteins, DNA, Single-Stranded, Humans, Mutation, Protein Binding, Recombination, Genetic, Sequence Homology, Nucleic Acid, Substrate Specificity, Cell Cycle Proteins, Amino Acid Motifs, BRCA2 Protein, Chromosomal Instability, Apoptosis Regulatory Proteins, Rad51 Recombinase, Replication Protein A
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