Stabilization of RAD51 nucleoprotein filaments by the C-terminal region of BRCA2

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Abstract

The human breast cancer susceptibility gene BRCA2 is required for the regulation of RAD51-mediated homologous recombinational repair. BRCA2 interacts with RAD51 monomers, as well as nucleoprotein filaments, primarily though the conserved BRC motifs. The unrelated C-terminal region of BRCA2 also interacts with RAD51. Here we show that the BRCA2 C terminus interacts directly with RAD51 filaments, but not monomers, by binding an interface created by two adjacent RAD51 protomers. These interactions stabilize filaments so that they cannot be dissociated by association with BRC repeats. Interaction of the BRCA2 C terminus with the RAD51 filament causes a large movement of the flexible RAD51 N-terminal domain that is important in regulating filament dynamics. We suggest that interactions of the BRCA2 C-terminal region with RAD51 may facilitate efficient nucleation of RAD51 multimers on DNA and thereby stimulate recombination-mediated repair.

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Figure 1: Effect of BRC4 on RAD51-TR2 interactions.
Figure 2: Analysis of the interaction of the C-terminal region of BRCA2 with RAD51 mutants that do not self-associate.
Figure 3: Particle size analysis of RAD51 in the presence of BRCA2 peptides.
Figure 4: Visualization of RAD51–DNA nucleoprotein filaments formed in the presence of TR2.
Figure 5: TR2 binds nucleoprotein filaments made by the RAD5182–339 core.
Figure 6: BRC4 does not disrupt RAD51–DNA nucleoprotein filaments in the presence of TR2.

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Acknowledgements

We thank R. Court and D. Wigley for help with the DLS analyses, and our lab members for their comments. This work was supported by Cancer Research UK, the Breast Cancer Campaign, the EU DNA Repair Consortium, the Jeantet Foundation (S.C.W.) and US National Institutes of Health grant GM035269 (E.H.E.). F.E. is a recipient of a postdoctoral fellowship from the Human Frontiers Science Program and the Japanese Society for the Promotion of Science.

Author information

F.E. designed and carried out the biochemical studies. V.E.G. and X.Y. carried out the EM studies. E.H.E. did the image reconstructions. F.E. and S.C.W. were responsible for manuscript preparation.

Correspondence to Stephen C West.

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The authors declare no competing financial interests.

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Esashi, F., Galkin, V., Yu, X. et al. Stabilization of RAD51 nucleoprotein filaments by the C-terminal region of BRCA2. Nat Struct Mol Biol 14, 468–474 (2007) doi:10.1038/nsmb1245

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