53BP1 loss rescues BRCA1 deficiency and is associated with triple-negative and BRCA-mutated breast cancers

Abstract

Germ-line mutations in breast cancer 1, early onset (BRCA1) result in predisposition to breast and ovarian cancer. BRCA1-mutated tumors show genomic instability, mainly as a consequence of impaired recombinatorial DNA repair. Here we identify p53-binding protein 1 (53BP1) as an essential factor for sustaining the growth arrest induced by Brca1 deletion. Depletion of 53BP1 abrogates the ATM-dependent checkpoint response and G2 cell-cycle arrest triggered by the accumulation of DNA breaks in Brca1-deleted cells. This effect of 53BP1 is specific to BRCA1 function, as 53BP1 depletion did not alleviate proliferation arrest or checkpoint responses in Brca2-deleted cells. Notably, loss of 53BP1 partially restores the homologous-recombination defect of Brca1-deleted cells and reverts their hypersensitivity to DNA-damaging agents. We find reduced 53BP1 expression in subsets of sporadic triple-negative and BRCA-associated breast cancers, indicating the potential clinical implications of our findings.

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Figure 1: Inactivation of 53BP1 rescues proliferation defects and drug sensitivity of Brca1-null ES cells.
Figure 2: 53BP1 depletion rescues cell-cycle defects of Brca1-null ES cells.
Figure 3: 53BP1 depletion abrogates CHK2-mediated DNA-damage responses and rescues proliferation defects in Brca1-null but not Brca2-null MEFs.
Figure 4: 53BP1 depletion rescues RAD51 foci in Brca1-null cells.
Figure 5: 53BP1 expression is reduced in a subset of human BLC.

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Acknowledgements

We wish to thank M. Treur-Mulder for her help with targeting of the Brca1SCo allele, W. Wang and P. Liu (Wellcome Trust Sanger Institute) for their kind gift of the piggyBac transposon MSCV 5′-LTR transposon and the mPB transposase, E. de Bruin, E. Cuppen and M. Koudijs for help with PCR amplification of the piggyBac insertions, J. Kong and D. Sie for mapping of piggyBac insertions, K, Szuhai for multicolor fluorescence in situ hybridization karyotyping, S. Philipsen (Erasmus Univ.) for providing shRNA vectors, H. te Riele and P. Borst for their comments on the manuscript, E. Bilal and G. Bhanot for their help with analysis of microarray data and P. Heikkilä and K. Aittomäki for their help with the Finnish breast cancer data and sample collection. We are most grateful to the patients who provided clinical samples that were analyzed in this study. Work in the Jonkers laboratory is supported by the Dutch Cancer Society, the Netherlands Organization for Scientific Research and the European Community 7th Framework Program (EuroSyStem project). Work in the Tarsounas laboratory is supported by Cancer Research UK and Breast Cancer Campaign. Work in the Ganesan laboratory is supported by the US Department of Defense (A.A.), the US National Cancer Institute (S.G.), the Sidney Kimmel Foundation (S.G.) and the Breast Cancer Research Foundation (S.G.). B.G.H. is supported by the Breast Cancer Research Foundation and the US National Cancer Institute. Work in the Bartek laboratory is supported by the Danish Cancer Society, the Danish National Research Foundation, Vilhelm Pedersen and Hustrus Mindelegat, the Czech Ministry of Education and the European Community 7th Framework Program (projects GENICA and INFLA-CARE). Work in the Nevanlinna laboratory is supported by the Helsinki University Central Hospital Research Fund, the Finnish Cancer Society, the Academy of Finland (132473) and the Sigrid Juselius Foundation. D.J.A. is supported by Cancer Research UK and the Wellcome Trust.

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P.B., A.A., J. Bartek, M.T., S.G. and J.J. designed research; P.B., A.A., J.M.E., M.P., J. Bartkova, H.v.d.G., S.H., M.T., A.K., Q.Y., B.G.H., J.T., C.B., D.J.A. and H.N. performed research; R.D. contributed new reagents/analytical tools; P.B., A.A., J.M.E., M.P., J. Bartek, H.N., M.T., S.G. and J.J. analyzed data; P.B., A.A., J. Bartek, M.T., S.G. and J.J. wrote the paper.

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Correspondence to Madalena Tarsounas or Shridar Ganesan or Jos Jonkers.

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Bouwman, P., Aly, A., Escandell, J. et al. 53BP1 loss rescues BRCA1 deficiency and is associated with triple-negative and BRCA-mutated breast cancers. Nat Struct Mol Biol 17, 688–695 (2010). https://doi.org/10.1038/nsmb.1831

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