Skip to main content

Thank you for visiting nature.com. You are using a browser version with limited support for CSS. To obtain the best experience, we recommend you use a more up to date browser (or turn off compatibility mode in Internet Explorer). In the meantime, to ensure continued support, we are displaying the site without styles and JavaScript.

  • Article
  • Published:

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

Nature Structural & Molecular Biology volume 17, pages 688–695 (2010)Cite this article

Subjects

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.

Access through your institution
Buy or subscribe

This is a preview of subscription content, access via your institution

Access options

Access through your institution

Buy this article

  • Purchase on SpringerLink
  • Instant access to full article PDF
Buy now

Prices may be subject to local taxes which are calculated during checkout

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.

Similar content being viewed by others

References

  1. Venkitaraman, A.R. Cancer susceptibility and the functions of BRCA1 and BRCA2. Cell 108, 171–182 (2002).

    Article  CAS  PubMed  Google Scholar 

  2. Gudmundsdottir, K. & Ashworth, A. The roles of BRCA1 and BRCA2 and associated proteins in the maintenance of genomic stability. Oncogene 25, 5864–5874 (2006).

    Article  CAS  PubMed  Google Scholar 

  3. Boulton, S.J. Cellular functions of the BRCA tumour-suppressor proteins. Biochem. Soc. Trans. 34, 633–645 (2006).

    Article  CAS  PubMed  Google Scholar 

  4. Collins, N. et al. Consistent loss of the wild type allele in breast cancers from a family linked to the BRCA2 gene on chromosome 13q12–13. Oncogene 10, 1673–1675 (1995).

    CAS  PubMed  Google Scholar 

  5. Smith, S.A., Easton, D.F., Evans, D.G. & Ponder, B.A. Allele losses in the region 17q12–21 in familial breast and ovarian cancer involve the wild-type chromosome. Nat. Genet. 2, 128–131 (1992).

    Article  CAS  PubMed  Google Scholar 

  6. Johannsson, O.T. et al. Tumour biological features of BRCA1-induced breast and ovarian cancer. Eur. J. Cancer 33, 362–371 (1997).

    Article  CAS  PubMed  Google Scholar 

  7. Foulkes, W.D. et al. Germline BRCA1 mutations and a basal epithelial phenotype in breast cancer. J. Natl. Cancer Inst. 95, 1482–1485 (2003).

    Article  CAS  PubMed  Google Scholar 

  8. Sorlie, T. et al. Repeated observation of breast tumor subtypes in independent gene expression data sets. Proc. Natl. Acad. Sci. USA 100, 8418–8423 (2003).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  9. Turner, N., Tutt, A. & Ashworth, A. Hallmarks of 'BRCAness' in sporadic cancers. Nat. Rev. Cancer 4, 814–819 (2004).

    Article  CAS  PubMed  Google Scholar 

  10. Evers, B. & Jonkers, J. Mouse models of BRCA1 and BRCA2 deficiency: past lessons, current understanding and future prospects. Oncogene 25, 5885–5897 (2006).

    Article  CAS  PubMed  Google Scholar 

  11. Hakem, R., de la Pompa, J.L., Elia, A., Potter, J. & Mak, T.W. Partial rescue of Brca1 (5–6) early embryonic lethality by p53 or p21 null mutation. Nat. Genet. 16, 298–302 (1997).

    Article  CAS  PubMed  Google Scholar 

  12. Cao, L. et al. A selective requirement for 53BP1 in the biological response to genomic instability induced by Brca1 deficiency. Mol. Cell 35, 534–541 (2009).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  13. Wang, B., Matsuoka, S., Carpenter, P.B. & Elledge, S.J. 53BP1, a mediator of the DNA damage checkpoint. Science 298, 1435–1438 (2002).

    Article  CAS  PubMed  Google Scholar 

  14. Morales, J.C. et al. 53BP1 and p53 synergize to suppress genomic instability and lymphomagenesis. Proc. Natl. Acad. Sci. USA 103, 3310–3315 (2006).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  15. Ward, I.M. et al. 53BP1 cooperates with p53 and functions as a haploinsufficient tumor suppressor in mice. Mol. Cell. Biol. 25, 10079–10086 (2005).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  16. Xu, X. et al. Genetic interactions between tumor suppressors Brca1 and p53 in apoptosis, cell cycle and tumorigenesis. Nat. Genet. 28, 266–271 (2001).

    Article  CAS  PubMed  Google Scholar 

  17. Hakem, R. et al. The tumor suppressor gene Brca1 is required for embryonic cellular proliferation in the mouse. Cell 85, 1009–1023 (1996).

    Article  CAS  PubMed  Google Scholar 

  18. Liu, X. et al. Somatic loss of BRCA1 and p53 in mice induces mammary tumors with features of human BRCA1-mutated basal-like breast cancer. Proc. Natl. Acad. Sci. USA 104, 12111–12116 (2007).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  19. Hameyer, D. et al. Toxicity of ligand-dependent Cre recombinases and generation of a conditional Cre deleter mouse allowing mosaic recombination in peripheral tissues. Physiol. Genomics 31, 32–41 (2007).

    Article  CAS  PubMed  Google Scholar 

  20. Wang, W. et al. Chromosomal transposition of PiggyBac in mouse embryonic stem cells. Proc. Natl. Acad. Sci. USA 105, 9290–9295 (2008).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  21. Bartz, S.R. et al. Small interfering RNA screens reveal enhanced cisplatin cytotoxicity in tumor cells having both BRCA network and TP53 disruptions. Mol. Cell. Biol. 26, 9377–9386 (2006).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  22. Kastan, M.B. & Bartek, J. Cell-cycle checkpoints and cancer. Nature 432, 316–323 (2004).

    Article  CAS  PubMed  Google Scholar 

  23. Taylor, W.R. & Stark, G.R. Regulation of the G2/M transition by p53. Oncogene 20, 1803–1815 (2001).

    Article  CAS  PubMed  Google Scholar 

  24. Silver, D.P. & Livingston, D.M. Self-excising retroviral vectors encoding the Cre recombinase overcome Cre-mediated cellular toxicity. Mol. Cell 8, 233–243 (2001).

    Article  CAS  PubMed  Google Scholar 

  25. Jonkers, J. et al. Synergistic tumor suppressor activity of BRCA2 and p53 in a conditional mouse model for breast cancer. Nat. Genet. 29, 418–425 (2001).

    Article  CAS  PubMed  Google Scholar 

  26. Jazayeri, A. et al. ATM- and cell cycle-dependent regulation of ATR in response to DNA double-strand breaks. Nat. Cell Biol. 8, 37–45 (2006).

    Article  CAS  PubMed  Google Scholar 

  27. Bhattacharyya, A., Ear, U.S., Koller, B.H., Weichselbaum, R.R. & Bishop, D.K. The breast cancer susceptibility gene BRCA1 is required for subnuclear assembly of Rad51 and survival following treatment with the DNA cross-linking agent cisplatin. J. Biol. Chem. 275, 23899–23903 (2000).

    Article  CAS  PubMed  Google Scholar 

  28. te Riele, H., Robanus-Maandag, E. & Berns, A. Highly efficient gene targeting in embryonic stem cells through homologous recombination with isogenic DNA constructs. Proc. Natl. Acad. Sci. USA 89, 5128–5132 (1992).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  29. te Riele, H., Robanus-Maandag, E., Clarke, A., Hooper, M. & Berns, A. Consecutive inactivation of both alleles of the pim-1 proto-oncogene by homologous recombination in embryonic stem cells. Nature 348, 649–651 (1990).

    Article  CAS  PubMed  Google Scholar 

  30. Holstege, H. et al. High incidence of protein-truncating TP53 mutations in BRCA1-related breast cancer. Cancer Res. 69, 3625–3633 (2009).

    Article  CAS  PubMed  Google Scholar 

  31. Manie, E. et al. High frequency of TP53 mutation in BRCA1 and sporadic basal-like carcinomas but not in BRCA1 luminal breast tumors. Cancer Res. 69, 663–671 (2009).

    Article  CAS  PubMed  Google Scholar 

  32. Ludwig, T., Chapman, D.L., Papaioannou, V.E. & Efstratiadis, A. Targeted mutations of breast cancer susceptibility gene homologs in mice: lethal phenotypes of Brca1, Brca2, Brca1/Brca2, Brca1/p53, and Brca2/p53 nullizygous embryos. Genes Dev. 11, 1226–1241 (1997).

    Article  CAS  PubMed  Google Scholar 

  33. Wang, Y. et al. Gene-expression profiles to predict distant metastasis of lymph-node-negative primary breast cancer. Lancet 365, 671–679 (2005).

    Article  CAS  PubMed  Google Scholar 

  34. Alexe, G. et al. High expression of lymphocyte-associated genes in node-negative HER2+ breast cancers correlates with lower recurrence rates. Cancer Res. 67, 10669–10676 (2007).

    Article  CAS  PubMed  Google Scholar 

  35. Richardson, A.L. et al. X chromosomal abnormalities in basal-like human breast cancer. Cancer Cell 9, 121–132 (2006).

    Article  CAS  PubMed  Google Scholar 

  36. Scully, R. et al. Dynamic changes of BRCA1 subnuclear location and phosphorylation state are initiated by DNA damage. Cell 90, 425–435 (1997).

    Article  CAS  PubMed  Google Scholar 

  37. Bartek, J. & Hodny, Z. SUMO boosts the DNA damage response barrier against cancer. Cancer Cell 17, 9–11 (2010).

    Article  CAS  PubMed  Google Scholar 

  38. Galanty, Y. et al. Mammalian SUMO E3-ligases PIAS1 and PIAS4 promote responses to DNA double-strand breaks. Nature 462, 935–939 (2009).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  39. Morris, J.R. et al. The SUMO modification pathway is involved in the BRCA1 response to genotoxic stress. Nature 462, 886–890 (2009).

    Article  CAS  PubMed  Google Scholar 

  40. Deng, C.X. BRCA1: cell cycle checkpoint, genetic instability, DNA damage response and cancer evolution. Nucleic Acids Res. 34, 1416–1426 (2006).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  41. Ward, I.M., Minn, K., van Deursen, J. & Chen, J. p53 Binding protein 53BP1 is required for DNA damage responses and tumor suppression in mice. Mol. Cell. Biol. 23, 2556–2563 (2003).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  42. Bunting, S.F. et al. 53BP1 inhibits homologous recombination in Brca1-deficient cells by blocking resection of DNA breaks. Cell 141, 243–254 (2010).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  43. Shiotani, B. & Zou, L. Single-stranded DNA orchestrates an ATM-to-ATR switch at DNA breaks. Mol. Cell 33, 547–558 (2009).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  44. Silver, D.P. et al. Efficacy of neoadjuvant Cisplatin in triple-negative breast cancer. J. Clin. Oncol. 28, 1145–1153 (2010).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  45. Fong, P.C. et al. Inhibition of poly(ADP-ribose) polymerase in tumors from BRCA mutation carriers. N. Engl. J. Med. 361, 123–134 (2009).

    Article  CAS  PubMed  Google Scholar 

  46. Rottenberg, S. et al. High sensitivity of BRCA1-deficient mammary tumors to the PARP inhibitor AZD2281 alone and in combination with platinum drugs. Proc. Natl. Acad. Sci. USA 105, 17079–17084 (2008).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  47. Rottenberg, S. et al. Selective induction of chemotherapy resistance of mammary tumors in a conditional mouse model for hereditary breast cancer. Proc. Natl. Acad. Sci. USA 104, 12117–12122 (2007).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  48. Swisher, E.M. et al. Secondary BRCA1 mutations in BRCA1-mutated ovarian carcinomas with platinum resistance. Cancer Res. 68, 2581–2586 (2008).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  49. Shafee, N. et al. Cancer stem cells contribute to cisplatin resistance in Brca1/p53-mediated mouse mammary tumors. Cancer Res. 68, 3243–3250 (2008).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  50. Jacobs, J.J. et al. Senescence bypass screen identifies TBX2, which represses Cdkn2a (p19(ARF)) and is amplified in a subset of human breast cancers. Nat. Genet. 26, 291–299 (2000).

    Article  CAS  PubMed  Google Scholar 

  51. Tarsounas, M. et al. Telomere maintenance requires the RAD51D recombination/repair protein. Cell 117, 337–347 (2004).

    Article  CAS  PubMed  Google Scholar 

  52. Dirac, A.M. & Bernards, R. Reversal of senescence in mouse fibroblasts through lentiviral suppression of p53. J. Biol. Chem. 278, 11731–11734 (2003).

    Article  CAS  PubMed  Google Scholar 

  53. Palmero, I. & Serrano, M. Induction of senescence by oncogenic Ras. Methods Enzymol. 333, 247–256 (2001).

    Article  CAS  PubMed  Google Scholar 

  54. Tarsounas, M., Davies, D. & West, S.C. BRCA2-dependent and independent formation of RAD51 nuclear foci. Oncogene 22, 1115–1123 (2003).

    Article  CAS  PubMed  Google Scholar 

Download references

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.

Author information

Author notes

  1. Peter Bouwman, Amal Aly and Jose M Escandell: These authors contributed equally to this work.

Authors and Affiliations

  1. Division of Molecular Biology, The Netherlands Cancer Institute, Amsterdam, The Netherlands

    Peter Bouwman, Mark Pieterse, Hanneke van der Gulden, Sanne Hiddingh & Jos Jonkers

  2. Cancer Institute of New Jersey, New Brunswick, New Jersey, USA

    Amal Aly, Atul Kulkarni, Qifeng Yang, Bruce G Haffty & Shridar Ganesan

  3. Telomere and Genome Stability Group, The Cancer Research UK-MRC Gray Institute for Radiation Oncology and Biology, Oxford, UK

    Jose M Escandell, Maria Thanasoula & Madalena Tarsounas

  4. Institute of Cancer Biology and Centre for Genotoxic Stress Research, Danish Cancer Society, Copenhagen, Denmark

    Jirina Bartkova & Jiri Bartek

  5. Department of Obstetrics and Gynecology, Helsinki University Central Hospital, Helsinki, Finland

    Johanna Tommiska & Heli Nevanlinna

  6. Department of Oncology, Helsinki University Central Hospital, Helsinki, Finland

    Carl Blomqvist

  7. Department of Medical Oncology, Center of Molecular Oncologic Pathology, Dana-Farber Cancer Institute, Boston, Massachusetts, USA

    Ronny Drapkin

  8. Wellcome Trust Sanger Institute, Wellcome Trust Genome Campus Hinxton, Cambridge, UK

    David J Adams

  9. Institute of Molecular and Translational Medicine, Palacky University, Olomouc, Czech Republic

    Jiri Bartek

Authors
  1. Peter Bouwman
    View author publications

    You can also search for this author inPubMed Google Scholar

  2. Amal Aly
    View author publications

    You can also search for this author inPubMed Google Scholar

  3. Jose M Escandell
    View author publications

    You can also search for this author inPubMed Google Scholar

  4. Mark Pieterse
    View author publications

    You can also search for this author inPubMed Google Scholar

  5. Jirina Bartkova
    View author publications

    You can also search for this author inPubMed Google Scholar

  6. Hanneke van der Gulden
    View author publications

    You can also search for this author inPubMed Google Scholar

  7. Sanne Hiddingh
    View author publications

    You can also search for this author inPubMed Google Scholar

  8. Maria Thanasoula
    View author publications

    You can also search for this author inPubMed Google Scholar

  9. Atul Kulkarni
    View author publications

    You can also search for this author inPubMed Google Scholar

  10. Qifeng Yang
    View author publications

    You can also search for this author inPubMed Google Scholar

  11. Bruce G Haffty
    View author publications

    You can also search for this author inPubMed Google Scholar

  12. Johanna Tommiska
    View author publications

    You can also search for this author inPubMed Google Scholar

  13. Carl Blomqvist
    View author publications

    You can also search for this author inPubMed Google Scholar

  14. Ronny Drapkin
    View author publications

    You can also search for this author inPubMed Google Scholar

  15. David J Adams
    View author publications

    You can also search for this author inPubMed Google Scholar

  16. Heli Nevanlinna
    View author publications

    You can also search for this author inPubMed Google Scholar

  17. Jiri Bartek
    View author publications

    You can also search for this author inPubMed Google Scholar

  18. Madalena Tarsounas
    View author publications

    You can also search for this author inPubMed Google Scholar

  19. Shridar Ganesan
    View author publications

    You can also search for this author inPubMed Google Scholar

  20. Jos Jonkers
    View author publications

    You can also search for this author inPubMed Google Scholar

Contributions

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.

Corresponding authors

Correspondence to Madalena Tarsounas, Shridar Ganesan or Jos Jonkers.

Ethics declarations

Competing interests

The authors declare no competing financial interests.

Supplementary information

Supplementary Text and Figures

Supplementary Data, Supplementary Methods, Supplementary Tables 1 and 2 and Supplementary Figures 1–4 (PDF 3184 kb)

Rights and permissions

Reprints and permissions

About this article

Cite this article

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

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1038/nsmb.1831

Search

Advanced search

Quick links

Nature Briefing: Cancer

Sign up for the Nature Briefing: Cancer newsletter — what matters in cancer research, free to your inbox weekly.

Get what matters in cancer research, free to your inbox weekly. Sign up for Nature Briefing: Cancer