A region of human BRCA2 containing multiple BRC repeats promotes RAD51-mediated strand exchange.
Human BRCA2, a breast and ovarian cancer suppressor, binds to the DNA recombinase RAD51 through eight conserved BRC repeats, motifs of approximately 30 residues, dispersed across a large region of the protein. BRCA2 is essential for homologous recombination in vivo, but isolated BRC repeat peptides can prevent the assembly of RAD51 into active nucleoprotein filaments in vitro, suggesting a model in which BRCA2 sequesters RAD51 in undamaged cells, and promotes recombinase function after DNA damage. How BRCA2 might fulfill these dual functions is unclear. We have purified a fragment of human BRCA2 (BRCA2(BRC1-8)) with 1127 residues spanning all 8 BRC repeats but excluding the C-terminal DNA-binding domain (BRCA2(CTD)). BRCA2(BRC1-8) binds RAD51 nucleoprotein filaments in a ternary complex, indicating it may organize RAD51 on DNA. Human RAD51 is relatively ineffective in vitro at strand exchange between homologous DNA molecules unless non-physiological ions like NH4+ are present. In an ionic milieu more typical of the mammalian nucleus, BRCA2(BRCI-8) stimulates RAD51-mediated strand exchange, suggesting it may be an essential co-factor in vivo. Thus, the human BRC repeats, embedded within their surronding sequences as an eight-repeat unit, mediate homologous recombination independent of the BRCA2(CTD) through a previously unrecognized role in control of RAD51 activity.
Shivji
Mahmud K K
MK
Cancer Research UK Department of Oncology and The Medical Research Council Cancer Cell Unit, Hutchison/MRC Research Centre, University of Cambridge, Hills Road, Cambridge CB2 2XZ, UK.
Davies
Owen R
OR
Savill
Jane M
JM
Bates
Debbie L
DL
Pellegrini
Luca
L
Venkitaraman
Ashok R
AR
D024682:BRCA2 Protein / Q000737:chemistry* / Q000502:physiology*; D002942:Circular Dichroism; D004247:DNA / Q000378:metabolism; D006801:Humans; D010455:Peptides / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism; D051135:Rad51 Recombinase / Q000378:metabolism*; D011995:Recombination, Genetic; D020449:Repetitive Sequences, Amino Acid
D024682:BRCA2 Protein; D010455:Peptides; D004247:DNA; C495153:RAD51 protein, human; D051135:Rad51 Recombinase