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hCds1-mediated phosphorylation of BRCA1 regulates the DNA damage response

Abstract

Mutations in the BRCA1 (ref. 1) tumour suppressor gene are found in almost all of the families with inherited breast and ovarian cancers and about half of the families with only breast cancer2,3. Although the biochemical function of BRCA1 is not well understood, it is important for DNA damage repair4,5,6,7 and cell-cycle checkpoint8,9,10. BRCA1 exists in nuclear foci but is hyperphosphorylated and disperses after DNA damage11,12. It is not known whether BRCA1 phosphorylation and dispersion and its function in DNA damage response are related. In yeast the DNA damage response and the replication-block checkpoint are mediated partly through the Cds1 kinase family13,14,15,16,17,18,19,20. Here we report that the human Cds1 kinase (hCds1/Chk2)21,22,23 regulates BRCA1 function after DNA damage by phosphorylating serine 988 of BRCA1. We show that hCds1 and BRCA1 interact and co-localize within discrete nuclear foci but separate after gamma irradiation. Phosphorylation of BRCA1 at serine 988 is required for the release of BRCA1 from hCds1. This phosphorylation is also important for the ability of BRCA1 to restore survival after DNA damage in the BRCA1-mutated cell line HCC1937.

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Figure 1: Cds1 phosphorylates Ser 988 of BRCA1 in vitro.
Figure 2: Cds1 phosphorylates Ser 988 of human BRCA1 in vivo.
Figure 3: Cds1 and BRCA1 are localized to overlapping nuclear foci.
Figure 4: BRCA1–hCds1 interaction is sensitive to DNA damage.
Figure 5: Phosphorylation of Ser 988 is important for BRCA1 function.

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References

  1. Miki, Y. et al. A strong candidate for the breast and ovarian cancer susceptibility gene BRCA1. Science 266, 66– 71 (1994).

    Article  ADS  CAS  Google Scholar 

  2. Easton, D. F., Bishop, D. T., Ford, D. & Crockford, G. P. Genetic linkage analysis in familial breast and ovarian cancer: results from 214 families. The breast Cancer Linkage Consortium. Am. J. Hum. Genet. 52, 678–701 (1993).

    CAS  PubMed  PubMed Central  Google Scholar 

  3. Feunteun, J. & Lenoir, G. M. BRCA1, a gene involved in inherited predisposition to breast and ovarian cancer. Biochim. Biophys. Acta 1242, 177–180 ( 1996).

    PubMed  Google Scholar 

  4. Gowen, L. C., Avrutskaya, A. V., Latour, A M., Koller, B. H. & Leadon, S. A. BRCA1 required for transcription-coupled repair of oxidative DNA damage. Science 281, 1009–1012 (1998).

    Article  ADS  CAS  Google Scholar 

  5. Shen, S. X. et al. A targeted disruption of the murine Brca1 gene causes gamma-irradiation hypersensitivity and genetic instability. Oncogene 17, 3115–3124 (1998).

    Article  CAS  Google Scholar 

  6. Abbott, D. W. et al. BRCA1 expression restores radiation resistance in BRCA1-defective cancer cells through enhancement of transcription-coupled DNA repair. J. Biol. Chem. 274, 18808–18812 (1999).

    Article  CAS  Google Scholar 

  7. Moynahan, M. E., Chiu, J. W., Koller, B. H. & Jasin, M. Brca1 controls homology-directed DNA repair. Mol. Cell 4, 511–518 (1999).

    Article  CAS  Google Scholar 

  8. Somasundaram, K. et al. Arrest of the cell cycle by the tumour-suppressor BRCA1 requires the CDK-inhibitor p21WAF1/CiP1. Nature 389, 187–190 (1997).

    Article  ADS  CAS  Google Scholar 

  9. Larson, J. S., Tonkinson, J. L. & Lai, M. T. A BRCA1 mutant alters G2-M cell cycle control in human mammary epithelial cells. Cancer Res. 57, 3351–3355 (1997).

    CAS  PubMed  Google Scholar 

  10. Xu, X. et al. Centrosome amplification and a defective G2-M cell cycle checkpoint induce genetic instability in BRCA1 exon 11 isoform-deficient cells. Mol. Cell 3, 389–395 ( 1999).

    Article  CAS  Google Scholar 

  11. 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  Google Scholar 

  12. Thomas, J. E., Smith, M., Tonkinson, J. L., Rubinfeld, B. & Polakis, P. Induction of phosphorylation on BRCA1 during the cell cycle and after DNA damage. Cell Growth Differ. 8, 801–809 ( 1997).

    CAS  Google Scholar 

  13. Zheng, P. et al. SPK1 is an essential S-phase-specific gene of Saccharomyces cerevisiae that encodes a nuclear serine/threonine/tyrosine kinase. Mol. Cell. Biol. 13, 5829–5842 (1993).

    Article  CAS  Google Scholar 

  14. Allen, J. B., Zhou, Z., Siede, W., Friedberg, E. C. & Elledge, S. J. The SAD1/RAD53 protein kinase controls multiple checkpoints and DNA damage-induced transcription in yeast. Genes Dev. 8, 2401–2415 (1994).

    Article  CAS  Google Scholar 

  15. Murakami, H. & Okayama, H. A kinase from fission yeast responsible for blocking mitosis in S phase. Nature 374, 817–819 (1995).

    Article  ADS  CAS  Google Scholar 

  16. Sanchez, Y. et al. Regulation of RAD53 by the ATM-like kinases MEC1 and TEL1 in yeast cell cycle checkpoint pathways. Science 271 , 357–360 (1996).

    Article  ADS  CAS  Google Scholar 

  17. Sidorova, J. M. & Breeden, L. L. Rad53-dependent phosphorylation of Swi6 and down-regulation of CLN1 and CLN2 transcription occur in response to DNA damage in Saccharomyces cerevisiae. Genes Dev. 11, 3032–3045 (1997).

    Article  CAS  Google Scholar 

  18. Lindsay, H. D. et al. S-phase-specific activation of Cds1 kinase defines a subpathway of the checkpoint response in Schizosaccharomyces pombe. Genes Dev. 12, 382–395 ( 1998).

    Article  CAS  Google Scholar 

  19. Boddy, M. N., Furnari, B., Mondesert, O. & Russell, P. Replication checkpoint enforced by kinases Cds1 and Chk1. Nature 395, 507–510 ( 1998).

    Article  Google Scholar 

  20. Zeng, Y. et al. Replication checkpoint requires phosphorylation of the phosphatase Cdc25 by Cds1 or Chk1. Nature 395, 507– 510 (1998).

    Article  ADS  CAS  Google Scholar 

  21. Matsuoka, S., Huang, M. & Elledge, S. J. Linkage of ATM to cell cycle regulation by the Chk2 protein kinase. Science 282, 1893– 1897 (1998).

    Article  ADS  CAS  Google Scholar 

  22. Blasina, A. et al. A human homologue of the checkpoint kinase Cds1 directly inhibits Cdc25 phosphatase. Curr. Biol. 9, 1– 10 (1999).

    Article  CAS  Google Scholar 

  23. Brown, A. L. et al. A human Cds1-related kinase that functions downstream of ATM protein in the cellular response to DNA damage. Proc. Natl Acad. Sci. USA 96, 3745–3750 ( 1999).

    Article  ADS  CAS  Google Scholar 

  24. Zhong, Q. et al. Association of BRCA1 with the hRad50-hMre11-p95 complex and the DNA damage response. Science 285, 747– 750 (1999).

    Article  CAS  Google Scholar 

  25. Cortez, D., Wang, Y., Qin, J. & Elledge, S. J. Requirement of ATM-dependent phosphorylation of brca1 in the DNA damage response to double-strand breaks. Science 286, 1162– 1166 (1999).

    Article  CAS  Google Scholar 

  26. Ruffner, H., Jiang, W., Craig, A. G., Hunter, T. & Verma, I. M. BRCA1 is phosphorylated at serine 1497 in vivo at a cyclin-dependent kinase 2-phosphorylation site. Mol. Cell. Biol. 19, 4843–4854 ( 1999).

    Article  CAS  Google Scholar 

  27. Scully, R. et al. Association of BRCA1 with Rad51 in mitotic and meiotic cells. Cell 88, 265–275 (1997).

    Article  CAS  Google Scholar 

  28. Ziv, Y. et al. Recombinant ATM protein complements the cellular A-T phenotype. Oncogene 15, 159–167 (1997).

    Article  CAS  Google Scholar 

  29. Wilson, C. A. et al. Localization of human BRCA1 and its loss in high-grade, non-inherited breast carcinomas. Nature Genet. 21, 236 –240 (1999).

    Article  CAS  Google Scholar 

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Acknowledgements

We thank D. Livingston and R. Scully for the BRCA1 plasmids; Y. Shiloh for the AT cells; M. Kirby for flow cytometry; and Y. Xu and C. Combs for help with confocal microscopy. We also thank W. S. Lane, D. P. Kirby and K. A. Pierce of the Harvard Microchemistry facility for expertise in tandem mass spectrometry peptide sequencing.

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Correspondence to Jay H. Chung.

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Lee, JS., Collins, K., Brown, A. et al. hCds1-mediated phosphorylation of BRCA1 regulates the DNA damage response . Nature 404, 201–204 (2000). https://doi.org/10.1038/35004614

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