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:

BRCA1 tumour suppression occurs via heterochromatin-mediated silencing

Nature volume 477, pages 179–184 (2011)Cite this article

Subjects

Abstract

Mutations in the tumour suppressor gene BRCA1 lead to breast and/or ovarian cancer. Here we show that loss of Brca1 in mice results in transcriptional de-repression of the tandemly repeated satellite DNA. Brca1 deficiency is accompanied by a reduction of condensed DNA regions in the genome and loss of ubiquitylation of histone H2A at satellite repeats. BRCA1 binds to satellite DNA regions and ubiquitylates H2A in vivo. Ectopic expression of H2A fused to ubiquitin reverses the effects of BRCA1 loss, indicating that BRCA1 maintains heterochromatin structure via ubiquitylation of histone H2A. Satellite DNA de-repression was also observed in mouse and human BRCA1-deficient breast cancers. Ectopic expression of satellite DNA can phenocopy BRCA1 loss in centrosome amplification, cell-cycle checkpoint defects, DNA damage and genomic instability. We propose that the role of BRCA1 in maintaining global heterochromatin integrity accounts for many of its tumour suppressor functions.

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

Access options

Buy this article

  • Purchase on Springer Link
  • Instant access to full article PDF
Buy now

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

Figure 1: Brca1 deficiency impairs heterochromatin structure.
Figure 2: BRCA1 and its ubiquitin E3 ligase activity are required for gene silencing in constitutive heterochromatin.
Figure 3: Ubiquitylated histone H2A is directly involved in BRCA1-mediated heterochromatic silencing.
Figure 4: De-repression of satellite DNA transcription occurs in Brca1 -deficient breast cancers.
Figure 5: Ectopic expression of satellite DNA transcripts leads to genomic instability in human mammary epithelial cells.

Similar content being viewed by others

Accession codes

Primary accessions

Gene Expression Omnibus

Data deposits

Microarray data have been deposited in the GEO database under the accession number GSE6310 (brain BRCA1 conditional knockout).

References

  1. Tutt, A. & Ashworth, A. The relationship between the roles of BRCA genes in DNA repair and cancer predisposition. Trends Mol. Med. 8, 571–576 (2002)

    Article  CAS  Google Scholar 

  2. King, M. C., Marks, J. H. & Mandell, J. B. Breast and ovarian cancer risks due to inherited mutations in BRCA1 and BRCA2. Science 302, 643–646 (2003)

    Article  ADS  CAS  Google Scholar 

  3. Huen, M. S., Sy, S. M. & Chen, J. BRCA1 and its toolbox for the maintenance of genome integrity. Nature Rev. Mol. Cell Biol. 11, 138–148 (2010)

    Article  CAS  Google Scholar 

  4. Pageau, G. J., Hall, L. L., Ganesan, S., Livingston, D. M. & Lawrence, J. B. The disappearing Barr body in breast and ovarian cancers. Nature Rev. Cancer 7, 628–633 (2007)

    Article  CAS  Google Scholar 

  5. Lane, T. F. et al. Expression of Brca1 is associated with terminal differentiation of ectodermally and mesodermally derived tissues in mice. Genes Dev. 9, 2712–2722 (1995)

    Article  CAS  Google Scholar 

  6. Korhonen, L., Brannvall, K., Skoglosa, Y. & Lindholm, D. Tumor suppressor gene BRCA-1 is expressed by embryonic and adult neural stem cells and involved in cell proliferation. J. Neurosci. Res. 71, 769–776 (2003)

    Article  CAS  Google Scholar 

  7. Maison, C. & Almouzni, G. HP1 and the dynamics of heterochromatin maintenance. Nature Rev. Mol. Cell Biol. 5, 296–305 (2004)

    Article  CAS  Google Scholar 

  8. Xia, Y., Pao, G. M., Chen, H. W., Verma, I. M. & Hunter, T. Enhancement of BRCA1 E3 ubiquitin ligase activity through direct interaction with the BARD1 protein. J. Biol. Chem. 278, 5255–5263 (2003)

    Article  CAS  Google Scholar 

  9. Wang, H. et al. Role of histone H2A ubiquitination in Polycomb silencing. Nature 431, 873–878 (2004)

    Article  ADS  CAS  Google Scholar 

  10. Meneghini, M. D., Wu, M. & Madhani, H. D. Conserved histone variant H2A.Z protects euchromatin from the ectopic spread of silent heterochromatin. Cell 112, 725–736 (2003)

    Article  CAS  Google Scholar 

  11. Martens, J. H. et al. The profile of repeat-associated histone lysine methylation states in the mouse epigenome. EMBO J. 24, 800–812 (2005)

    Article  CAS  Google Scholar 

  12. Hashizume, R. et al. The RING heterodimer BRCA1-BARD1 is a ubiquitin ligase inactivated by a breast cancer-derived mutation. J. Biol. Chem. 276, 14537–14540 (2001)

    Article  CAS  Google Scholar 

  13. Reid, L. J. et al. E3 ligase activity of BRCA1 is not essential for mammalian cell viability or homology-directed repair of double-strand DNA breaks. Proc. Natl Acad. Sci. USA 105, 20876–20881 (2008)

    Article  ADS  CAS  Google Scholar 

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

    Article  CAS  Google Scholar 

  15. Sankaran, S., Starita, L. M., Groen, A. C., Ko, M. J. & Parvin, J. D. Centrosomal microtubule nucleation activity is inhibited by BRCA1-dependent ubiquitination. Mol. Cell. Biol. 25, 8656–8668 (2005)

    Article  CAS  Google Scholar 

  16. Matsui, S. I., Seon, B. K. & Sandberg, A. A. Disappearance of a structural chromatin protein A24 in mitosis: implications for molecular basis of chromatin condensation. Proc. Natl Acad. Sci. USA 76, 6386–6390 (1979)

    Article  ADS  CAS  Google Scholar 

  17. Kallin, E. M. et al. Genome-wide uH2A localization analysis highlights Bmi1-dependent deposition of the mark at repressed genes. PLoS Genet. 5, e1000506 (2009)

    Article  Google Scholar 

  18. Zhou, W. et al. Histone H2A monoubiquitination represses transcription by inhibiting RNA polymerase II transcriptional elongation. Mol. Cell 29, 69–80 (2008)

    Article  Google Scholar 

  19. Doil, C. et al. RNF168 binds and amplifies ubiquitin conjugates on damaged chromosomes to allow accumulation of repair proteins. Cell 136, 435–446 (2009)

    Article  CAS  Google Scholar 

  20. Stewart, G. S. et al. The RIDDLE syndrome protein mediates a ubiquitin-dependent signaling cascade at sites of DNA damage. Cell 136, 420–434 (2009)

    Article  CAS  Google Scholar 

  21. Scheuermann, J. C. et al. Histone H2A deubiquitinase activity of the Polycomb repressive complex PR-DUB. Nature 465, 243–247 (2010)

    Article  ADS  CAS  Google Scholar 

  22. Jensen, D. E. et al. BAP1: a novel ubiquitin hydrolase which binds to the BRCA1 RING finger and enhances BRCA1-mediated cell growth suppression. Oncogene 16, 1097–1112 (1998)

    Article  CAS  Google Scholar 

  23. Ruffner, H., Joazeiro, C. A. P., Hemmati, D., Hunter, T. & Verma, I. M. Cancer-predisposing mutations within the RING domain of BRCA1: Loss of ubiquitin protein ligase activity and protection from radiation hypersensitivity. Proc. Natl Acad. Sci. USA 98, 5134–5139 (2001)

    Article  ADS  CAS  Google Scholar 

  24. Ting, D. T. et al. Aberrant overexpression of satellite repeats in pancreatic and other epithelial cancers. Science 331, 593–596 (2011)

    Article  ADS  CAS  Google Scholar 

  25. Palmer, T. D., Markakis, E. A., Willhoite, A. R., Safar, F. & Gage, F. H. Fibroblast growth factor-2 activates a latent neurogenic program in neural stem cells from diverse regions of the adult CNS. J. Neurosci. 19, 8487–8497 (1999)

    Article  CAS  Google Scholar 

  26. Nakashima, K. et al. Synergistic signaling in fetal brain by STAT3-Smad1 complex bridged by p300. Science 284, 479–482 (1999)

    Article  ADS  CAS  Google Scholar 

  27. van Es, J. H. et al. Wnt signalling induces maturation of Paneth cells in intestinal crypts. Nature Cell Biol. 7, 381–386 (2005)

    Article  CAS  Google Scholar 

  28. Lie, D. C. et al. Wnt signalling regulates adult hippocampal neurogenesis. Nature 437, 1370–1375 (2005)

    Article  ADS  CAS  Google Scholar 

  29. Huang, J. et al. Lsh, an epigenetic guardian of repetitive elements. Nucleic Acids Res. 32, 5019–5028 (2004)

    Article  CAS  Google Scholar 

Download references

Acknowledgements

We thank B. Miller for assistance in culturing NPCs; E. Ke for discussion and analysis of Affymetrix data; A. Yanai for the BRCA1 targeting shRNA construct; C. Lilley for assistance with western blotting; and Z. You for assistance with the LI-COR Odyssey Infrared Imaging System. We thank A. Berns for his sustained interest in this work and providing mutant mice and materials, and M. Vidal for providing mouse embryo fibroblasts containing a conditional deletion allele of Ring1B. Q.Z. was supported by the California Breast Cancer Research Program and Ruth L. Kirschtein National Research Service Award. G.M.P. was supported by a fellowship of the California Institute of Regenerative Medicine. H.S. is a recipient of ASPET-Merck fellowship. I.M.V. is an American Cancer Society Professor of Molecular Biology, and holds the Irwin and Joan Jacobs Chair in Exemplary Life Science. This work was supported in part by grants from the NIH, Ipsen/Biomeasure, Sanofi Aventis, and the H.N. and Frances C. Berger Foundation. F.H.G. is supported by NIH NS52842, NS50217 and the Lookout Fund.

Author information

Author notes

  1. Alexis M. Huynh & Hoonkyo Suh

    Present address: Present addresses: Department of Medicine, Beth Israel Medical Center, Albert Einstein College of Medicine, New York, New York 10461, USA (A.M.H.); Stem Cells and Regenerative Medicine, Lerner Research Institute/Cleveland Clinic, 9500 Euclid Avenue, Cleveland, Ohio 44195, USA (H.S.).,

  2. Quan Zhu and Gerald M. Pao: These authors contributed equally to this work.

Authors and Affiliations

  1. Laboratory of Genetics, The Salk Institute for Biological Studies, La Jolla, 92037, California, USA

    Quan Zhu, Gerald M. Pao, Alexis M. Huynh, Hoonkyo Suh, Nina Tonnu, Fred H. Gage & Inder M. Verma

  2. Department of Pathology, the Netherlands Cancer Institute, 1066 CX, Amsterdam, The Netherlands

    Petra M. Nederlof

Authors
  1. Quan Zhu
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Gerald M. Pao
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Alexis M. Huynh
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Hoonkyo Suh
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Nina Tonnu
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Petra M. Nederlof
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Fred H. Gage
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. Inder M. Verma
    View author publications

    You can also search for this author in PubMed Google Scholar

Contributions

Q.Z. generated and Q.Z., N.T. and G.M.P. maintained all the knockout mice. G.M.P. and Q.Z. made the initial heterochromatin observation. Q.Z. and A.M.H. performed confocal microscopy experiments. G.M.P., Q.Z. and N.T. performed ChIP experiments. RNA isolation and microarray experiments were performed by Q.Z., G.M.P. and N.T. Microdissection of murine brains were performed by G.M.P. and A.M.H. under the guidance of F.H.G. G.M.P. designed the H2A–ubiquitin fusion experiments that were performed by Q.Z., G.M.P. and N.T. Satellite RNA experiments were designed by G.M.P. and Q.Z. and performed by Q.Z., G.M.P. and N.T. H.S. established the embryonic neural stem cell isolation and culture. P.M.N. obtained, isolated and curated the clinical patient samples, which were analysed by Q.Z., G.M.P. and N.T. All other experiments were performed by Q.Z., G.M.P. and N.T. All experiments and experimental design was performed under the supervision of I.M.V. G.M.P., Q.Z. and I.M.V. wrote the manuscript.

Corresponding authors

Correspondence to Gerald M. Pao or Inder M. Verma.

Ethics declarations

Competing interests

The authors declare no competing financial interests.

Supplementary information

Supplementary Figures

The file contains Supplementary Figures 1-14 with legends. (PDF 1249 kb)

PowerPoint slides

Rights and permissions

Reprints and permissions

About this article

Cite this article

Zhu, Q., Pao, G., Huynh, A. et al. BRCA1 tumour suppression occurs via heterochromatin-mediated silencing. Nature 477, 179–184 (2011). https://doi.org/10.1038/nature10371

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1038/nature10371

This article is cited by

Comments

By submitting a comment you agree to abide by our Terms and Community Guidelines. If you find something abusive or that does not comply with our terms or guidelines please flag it as inappropriate.

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