Interaction with the BRCA2 C terminus protects RAD51–DNA filaments from disassembly by BRC repeats

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  • A Corrigendum to this article was published on 01 July 2007

Abstract

BRCA2 has an essential function in DNA repair by homologous recombination, interacting with RAD51 via short motifs in the middle and at the C terminus of BRCA2. Here, we report that a conserved 36-residue sequence of human BRCA2 encoded by exon 27 (BRCA2Exon27) interacts with RAD51 through the specific recognition of oligomerized RAD51 ATPase domains. BRCA2Exon27 binding stabilizes the RAD51 nucleoprotein filament against disassembly by BRC repeat 4. The protection is specific for RAD51 filaments formed on single-stranded DNA and is lost when BRCA2Exon27 is phosphorylated on Ser3291. We propose that productive recombination results from the functional balance between the different RAD51-binding modes of the BRC repeat and exon 27 regions of BRCA2. Our results further suggest a mechanism in which CDK phosphorylation of BRCA2Exon27 at the G2-M transition alters the balance in favor of RAD51 filament disassembly, thus terminating recombination.

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Figure 1: Structure and function of BRCA2 and RAD51 proteins.
Figure 2: BRCA2Exon27 binds oligomeric RAD51.
Figure 3: Cross-linking of RAD51, RAD51–BRC4 and RAD51–BRCA2Exon27 complexes.
Figure 4: BRCA2Exon27 binding requires an oligomeric RAD51 assembly.
Figure 5: BRCA2Exon27 binds RAD51 nucleoprotein filaments.
Figure 6: BRCA2Exon27 protects RAD51 nucleoprotein filaments.
Figure 7: EM of RAD51 nucleoprotein filaments.
Figure 8: BRCA2Exon27 protection is specific for ssDNA and independent of ATP hydrolysis.

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Acknowledgements

We thank T. Blundell, S. Klinge, M. Shivji, E. Rajendra, N. Ayoub and A. Venkitaraman for comments and advice, N. Allcott and T. Yamamoto for their contributions in the early stages of this work, and J. Skepper (University of Cambridge) for assistance with EM. This work was supported by a Wellcome Trust Senior Fellowship in basic biomedical sciences to L.P. and a Biotechnology and Biological Sciences Research Council PhD studentship to O.R.D.

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O.R.D. and L.P. designed the experiments and interpreted the data; O.R.D. performed all experiments.

Correspondence to Luca Pellegrini.

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The authors declare no competing financial interests.

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Supplementary Methods

Preparation of DNA substrates. (PDF 43 kb)

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Davies, O., Pellegrini, L. Interaction with the BRCA2 C terminus protects RAD51–DNA filaments from disassembly by BRC repeats. Nat Struct Mol Biol 14, 475–483 (2007) doi:10.1038/nsmb1251

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