Abstract
The breast cancer susceptibility protein, BRCA2, is essential for recombinational DNA repair. BRCA2 delivers RAD51 to double-stranded DNA (dsDNA) breaks through interaction with eight conserved, ∼35 amino acid motifs, the BRC repeats. Here we show that the solitary BRC4 promotes assembly of RAD51 onto single-stranded DNA (ssDNA), but not dsDNA, to stimulate DNA strand exchange. BRC4 acts by blocking ATP hydrolysis and thereby maintaining the active ATP-bound form of the RAD51-ssDNA filament. Single-molecule visualization shows that BRC4 does not disassemble RAD51-dsDNA filaments but rather blocks nucleation of RAD51 onto dsDNA. Furthermore, this behavior is manifested by a domain of BRCA2 comprising all eight BRC repeats. These results establish that the BRC repeats modulate RAD51-DNA interaction in two opposing but functionally reinforcing ways: targeting active RAD51 to ssDNA and prohibiting RAD51 nucleation onto dsDNA. Thus, BRCA2 recruits RAD51 to DNA breaks and, we propose, the BRC repeats regulate DNA-binding selectivity.
Original language | English (US) |
---|---|
Pages (from-to) | 1032-1043 |
Number of pages | 12 |
Journal | Cell |
Volume | 136 |
Issue number | 6 |
DOIs | |
State | Published - Mar 20 2009 |
Fingerprint
Keywords
- DNA
ASJC Scopus subject areas
- Biochemistry, Genetics and Molecular Biology(all)
Cite this
The BRC Repeats of BRCA2 Modulate the DNA-Binding Selectivity of RAD51. / Carreira, Aura; Hilario, Jovencio; Amitani, Ichiro; Baskin, Ronald J.; Shivji, Mahmud K K; Venkitaraman, Ashok R.; Kowalczykowski, Stephen C.
In: Cell, Vol. 136, No. 6, 20.03.2009, p. 1032-1043.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - The BRC Repeats of BRCA2 Modulate the DNA-Binding Selectivity of RAD51
AU - Carreira, Aura
AU - Hilario, Jovencio
AU - Amitani, Ichiro
AU - Baskin, Ronald J.
AU - Shivji, Mahmud K K
AU - Venkitaraman, Ashok R.
AU - Kowalczykowski, Stephen C.
PY - 2009/3/20
Y1 - 2009/3/20
N2 - The breast cancer susceptibility protein, BRCA2, is essential for recombinational DNA repair. BRCA2 delivers RAD51 to double-stranded DNA (dsDNA) breaks through interaction with eight conserved, ∼35 amino acid motifs, the BRC repeats. Here we show that the solitary BRC4 promotes assembly of RAD51 onto single-stranded DNA (ssDNA), but not dsDNA, to stimulate DNA strand exchange. BRC4 acts by blocking ATP hydrolysis and thereby maintaining the active ATP-bound form of the RAD51-ssDNA filament. Single-molecule visualization shows that BRC4 does not disassemble RAD51-dsDNA filaments but rather blocks nucleation of RAD51 onto dsDNA. Furthermore, this behavior is manifested by a domain of BRCA2 comprising all eight BRC repeats. These results establish that the BRC repeats modulate RAD51-DNA interaction in two opposing but functionally reinforcing ways: targeting active RAD51 to ssDNA and prohibiting RAD51 nucleation onto dsDNA. Thus, BRCA2 recruits RAD51 to DNA breaks and, we propose, the BRC repeats regulate DNA-binding selectivity.
AB - The breast cancer susceptibility protein, BRCA2, is essential for recombinational DNA repair. BRCA2 delivers RAD51 to double-stranded DNA (dsDNA) breaks through interaction with eight conserved, ∼35 amino acid motifs, the BRC repeats. Here we show that the solitary BRC4 promotes assembly of RAD51 onto single-stranded DNA (ssDNA), but not dsDNA, to stimulate DNA strand exchange. BRC4 acts by blocking ATP hydrolysis and thereby maintaining the active ATP-bound form of the RAD51-ssDNA filament. Single-molecule visualization shows that BRC4 does not disassemble RAD51-dsDNA filaments but rather blocks nucleation of RAD51 onto dsDNA. Furthermore, this behavior is manifested by a domain of BRCA2 comprising all eight BRC repeats. These results establish that the BRC repeats modulate RAD51-DNA interaction in two opposing but functionally reinforcing ways: targeting active RAD51 to ssDNA and prohibiting RAD51 nucleation onto dsDNA. Thus, BRCA2 recruits RAD51 to DNA breaks and, we propose, the BRC repeats regulate DNA-binding selectivity.
KW - DNA
UR - http://www.scopus.com/inward/record.url?scp=62149104415&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=62149104415&partnerID=8YFLogxK
U2 - 10.1016/j.cell.2009.02.019
DO - 10.1016/j.cell.2009.02.019
M3 - Article
C2 - 19303847
AN - SCOPUS:62149104415
VL - 136
SP - 1032
EP - 1043
JO - Cell
JF - Cell
SN - 0092-8674
IS - 6
ER -