The BRC Repeats of BRCA2 Modulate the DNA-Binding Selectivity of RAD51

Aura Carreira, Jovencio Hilario, Ichiro Amitani, Ronald J. Baskin, Mahmud K K Shivji, Ashok R. Venkitaraman, Stephen C. Kowalczykowski

Research output: Contribution to journal › Article

134 Citations (Scopus)

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 languageEnglish (US)
Pages (from-to)1032-1043
Number of pages12
JournalCell
Volume136
Issue number6
DOIs
StatePublished - Mar 20 2009

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Single-Stranded DNA
DNA
BRCA2 Protein
Nucleation
Adenosine Triphosphate
Amino Acid Motifs
Hydrolysis
Repair
Visualization
Breast Neoplasms
Amino Acids
Molecules
Haemophilus influenzae type b-polysaccharide vaccine-diphtheria toxoid conjugate

Keywords

  • DNA

ASJC Scopus subject areas

  • Biochemistry, Genetics and Molecular Biology(all)

Cite this

Carreira, A., Hilario, J., Amitani, I., Baskin, R. J., Shivji, M. K. K., Venkitaraman, A. R., & Kowalczykowski, S. C. (2009). The BRC Repeats of BRCA2 Modulate the DNA-Binding Selectivity of RAD51. Cell, 136(6), 1032-1043. https://doi.org/10.1016/j.cell.2009.02.019

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

Carreira, A, Hilario, J, Amitani, I, Baskin, RJ, Shivji, MKK, Venkitaraman, AR & Kowalczykowski, SC 2009, 'The BRC Repeats of BRCA2 Modulate the DNA-Binding Selectivity of RAD51', Cell, vol. 136, no. 6, pp. 1032-1043. https://doi.org/10.1016/j.cell.2009.02.019
Carreira A, Hilario J, Amitani I, Baskin RJ, Shivji MKK, Venkitaraman AR et al. The BRC Repeats of BRCA2 Modulate the DNA-Binding Selectivity of RAD51. Cell. 2009 Mar 20;136(6):1032-1043. https://doi.org/10.1016/j.cell.2009.02.019
Carreira, Aura ; Hilario, Jovencio ; Amitani, Ichiro ; Baskin, Ronald J. ; Shivji, Mahmud K K ; Venkitaraman, Ashok R. ; Kowalczykowski, Stephen C. / The BRC Repeats of BRCA2 Modulate the DNA-Binding Selectivity of RAD51. In: Cell. 2009 ; Vol. 136, No. 6. pp. 1032-1043.
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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.",
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AU - Shivji, Mahmud K K

AU - Venkitaraman, Ashok R.

AU - Kowalczykowski, Stephen C.

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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.

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