A small-molecule inhibitor of the BRCA2-RAD51 interaction modulates RAD51 assembly and potentiates DNA damage-induced cell death

• Published in: Cell chemical biology Vol. 28; no. 6; pp. 835 - 847.e5
• Main Authors: Scott, Duncan E., Francis-Newton, Nicola J., Marsh, May E., Coyne, Anthony G., Fischer, Gerhard, Moschetti, Tommaso, Bayly, Andrew R., Sharpe, Timothy D., Haas, Kalina T., Barber, Lorraine, Valenzano, Chiara R., Srinivasan, Rajavel, Huggins, David J., Lee, Miyoung, Emery, Amy, Hardwick, Bryn, Ehebauer, Matthias, Dagostin, Claudio, Esposito, Alessandro, Pellegrini, Luca, Perrior, Trevor, McKenzie, Grahame, Blundell, Tom L., Hyvönen, Marko, Skidmore, John, Venkitaraman, Ashok R., Abell, Chris
• Format: Journal Article
• Language: English
• Published: United States Elsevier Ltd 17.06.2021; Cell Press
• Subjects: BRCA2; BRCA2 Protein - antagonists & inhibitors; BRCA2 Protein - chemistry; BRCA2 Protein - metabolism; cancer therapy; Cell Death - drug effects; Crystallography, X-Ray; DNA Damage; DNA repair; homologous recombination; Humans; Models, Molecular; Molecular Conformation; Protein Binding - drug effects; protein-protein interaction inhibition; RAD51; RAD51 inhibitor; Rad51 Recombinase - antagonists & inhibitors; Rad51 Recombinase - chemistry; Rad51 Recombinase - metabolism; radiosensitizer; Small Molecule Libraries - chemical synthesis; Small Molecule Libraries - chemistry; Small Molecule Libraries - pharmacology; structure-guided drug discovery; Tumor Cells, Cultured; cancer therapy; RAD51; homologous recombination; RAD51 inhibitor; structure-guided drug discovery; radiosensitizer; BRCA2; DNA repair; protein-protein interaction inhibition
• ISSN: 2451-9456; 2451-9456; 2451-9448
• DOI: 10.1016/j.chembiol.2021.02.006

BRCA2 controls RAD51 recombinase during homologous DNA recombination (HDR) through eight evolutionarily conserved BRC repeats, which individually engage RAD51 via the motif Phe-x-x-Ala. Using structure-guided molecular design, templated on a monomeric thermostable chimera between human RAD51 and archaeal RadA, we identify CAM833, a 529 Da orthosteric inhibitor of RAD51:BRC with a Kd of 366 nM. The quinoline of CAM833 occupies a hotspot, the Phe-binding pocket on RAD51 and the methyl of the substituted α-methylbenzyl group occupies the Ala-binding pocket. In cells, CAM833 diminishes formation of damage-induced RAD51 nuclear foci; inhibits RAD51 molecular clustering, suppressing extended RAD51 filament assembly; potentiates cytotoxicity by ionizing radiation, augmenting 4N cell-cycle arrest and apoptotic cell death and works with poly-ADP ribose polymerase (PARP)1 inhibitors to suppress growth in BRCA2-wildtype cells. Thus, chemical inhibition of the protein-protein interaction between BRCA2 and RAD51 disrupts HDR and potentiates DNA damage-induced cell death, with implications for cancer therapy. [Display omitted] •CAM833 inhibits the protein-protein interaction between RAD51 and BRCA2•Crystal structure shows CAM833 binds RAD51 at the same site as the BRCA2 FxxA motif•CAM833 blocks formation of RAD51 foci and filaments, preventing DNA repair•CAM833 potentiates cytotoxicity by IR and synergises with PARP1 inhibitors Scott et al. describe CAM833, an inhibitor of DNA recombinase RAD51, that binds at the site normally occupied by the BRC repeats of the BRCA2 protein. Through blocking this protein-protein interaction, CAM833 is able to prevent RAD51-mediated homologous recombination and thus potentiate cancer cells to damage by radiation or PARP1 inhibition.