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Checkpoint activation in response to double-strand breaks requires the Mre11/Rad50/Xrs2 complex

Nature Cell Biology volume 3, pages 844–847 (2001)Cite this article

Abstract

Studies of human Nijmegen breakage syndrome (NBS) cells have led to the proposal that the Mre11/Rad50/ NBS1 complex, which is involved in the repair of DNA double-strand breaks (DSBs), might also function in activating the DNA damage checkpoint pathways after DSBs occur1,2. We have studied the role of the homologous budding yeast complex, Mre11/Rad50/Xrs2, in checkpoint activation in response to DSB-inducing agents. Here we show that this complex is required for phosphorylation and activation of the Rad53 and Chk1 checkpoint kinases specifically in response to DSBs. Consistent with defective Rad53 activation, we observed defective cell-cycle delays after induction of DSBs in the absence of Mre11. Furthermore, after γ-irradiation phosphorylation of Rad9, which is an early event in checkpoint activation, is also dependent on Mre11. All three components of the Mre11/Rad50/Xrs2 complex are required for activation of Rad53, however, the Ku80, Rad51 or Rad52 proteins, which are also involved in DSB repair, are not. Thus, the integrity of the Mre11/Rad50/Xrs2 complex is specifically required for checkpoint activation after the formation of DSBs.

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Figure 1: Mre11 is involved in the DNA damage-dependent phosphorylation of Rad53 in response to γ-irradiation.
Figure 2: Analysis of transient cell-cycle delay in response to DSBs in mre11Δ cells.
Figure 3: Activation of both Chk1 and Rad9 after γ-irradiation requires Mre11.
Figure 4: The integrity of the Mre11 complex, but not Ku80, Rad51 nor Rad52, is required for checkpoint activation.

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References

  1. Haber, J. E. Cell 95, 583–586 (1998).

    Article  CAS  Google Scholar 

  2. Petrini, J. Am. J. Hum. Genet. 64, 1264–1269 (1999).

    Article  CAS  Google Scholar 

  3. Sanchez, Y. et al. Science 271, 357–360 (1996).

    Article  CAS  Google Scholar 

  4. Sun, Z., Fay, D. S., Marini, F., Foiani, M. & Stern, D. F. Genes Dev. 10, 395–406 (1996).

    Article  CAS  Google Scholar 

  5. de la Torre-Ruiz, M.-A., Green, C. M. & Lowndes, N. F. EMBO J. 17, 2687–2698 (1998).

    Article  CAS  Google Scholar 

  6. Pellicioli, A. et al. EMBO J. 18, 6561–6572 (1999).

    Article  CAS  Google Scholar 

  7. Paulovich, A. G. & Hartwell, L. H. Cell 82, 841–847 (1995).

    Article  CAS  Google Scholar 

  8. Aboussekhra, A. et al. EMBO J. 15, 3912–3922 (1996).

    Article  CAS  Google Scholar 

  9. Sanchez, Y. et al. Science 286, 1166–1171 (1999).

    Article  CAS  Google Scholar 

  10. Emili, A. Mol. Cell 2, 183–189 (1998).

    Article  CAS  Google Scholar 

  11. Vialard, J. E., Gilbert, C. S., Green, C. M. & Lowndes, N. F. EMBO J. 17, 5679–5688 (1998).

    Article  CAS  Google Scholar 

  12. Longhese, M. P. et al. EMBO J. 16, 5216–5226 (1997).

    Article  CAS  Google Scholar 

  13. Paciotti, V., Lucchini, G., Plevani, P. & Longhese, M. P. EMBO J. 17, 4199–4209 (1998).

    Article  CAS  Google Scholar 

  14. Paciotti, V., Clerici, M., Lucchini, G. & Longhese, M. P. Genes Dev. 14, 2046–2059 (2000).

    CAS  PubMed  PubMed Central  Google Scholar 

  15. Weinert, T. Curr. Opin. Genet. Dev. 8, 185–193 (1998).

    Article  CAS  Google Scholar 

  16. Lowndes, N. F. & Murguia, J. R. Curr. Opin. Genet. Dev. 10, 17–25 (2000).

    Article  CAS  Google Scholar 

  17. Usui, T. et al. Cell 95, 705–716 (1998).

    Article  CAS  Google Scholar 

  18. Nelms, B. E., Maser, R. S., Mackay, J. F., Lagally, M. G. & Petrini, J. H. Science 280, 590–590 (1998).

    Article  CAS  Google Scholar 

  19. Neecke, H., Lucchini, G. & Longhese, M. P. EMBO J. 18, 4485–4497 (1999).

    Article  CAS  Google Scholar 

  20. Carney, J. P. et al. Cell 93, 477–486 (1998).

    Article  CAS  Google Scholar 

  21. Stewart, G. S. et al. Cell 99, 577–587. (1999).

    Article  CAS  Google Scholar 

  22. Matsuura, K. et al. Biochem. Biophys. Res. Commun. 242, 602–607 (1998).

    Article  CAS  Google Scholar 

  23. Yamazaki, V., Wegner, R.-D. & Kirchgessner, C. U. Cancer Res. 58, 2316–2322 (1998).

    CAS  PubMed  Google Scholar 

  24. Antoccia, A. et al. Int. J. Radiat. Biol. 75, 583–591 (1999).

    Article  CAS  Google Scholar 

  25. Girard, P. et al. Cancer Res. 60, 4881–4888 (2000).

    CAS  PubMed  Google Scholar 

  26. Antoccia, A., Ricordy, R., Maraschio, P., Prudente, S. & Tanzarella, C. Int. J. Radiat. Biol. 71, 41–49 (1997).

    Article  CAS  Google Scholar 

  27. Pincheira, J., Bravo, M. & Santos, M. Clin. Genet. 53, 262–267 (1998).

    Article  CAS  Google Scholar 

  28. Xiao, Y. & Weaver, D. T. Nucleic Acids Res. 25, 2985–2991 (1997).

    Article  CAS  Google Scholar 

  29. Luo, G. et al. Proc. Natl Acad. Sci. USA 96, 7376–7381 (1999).

    Article  CAS  Google Scholar 

  30. Zhu, J., Petersen, S., Tessarollo, L. & Nussenzweig, A. Curr. Biol. 11, 105–109 (2001).

    Article  CAS  Google Scholar 

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Acknowledgements

We thank J. Diffley, S. Jackson and M. P. Longhese for yeast strains and plasmids; A. Verreault, S. Jackson and S. West for critical reading of the manuscript. We are indebted to C. Green and J. Murguia for many stimulating discussions. We also thank B. Sedgewick and A. Verreault for their generosity. M.G. was supported in part by La Ligue Nationale contre le Cancer, Paris.

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Author notes

  1. Muriel Grenon

    Present address: Wellcome/CRC Institute, Tennis Court Road, Cambridge, CB2 1QR, UK

Authors and Affiliations

  1. ICRF Clare Hall Laboratories, CDC Laboratory, Blanche Lane, South Mimms, Potters Bar, EN6 3LD, Hertfordshire, UK

    Muriel Grenon, Chris Gilbert & Noel F. Lowndes

  2. Department of Biochemistry, National University of Ireland Galway, University Road, Galway, Ireland

    Noel F. Lowndes

Authors
  1. Muriel Grenon
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  2. Chris Gilbert
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Corresponding author

Correspondence to Noel F. Lowndes.

Supplementary information

Supplementary figures and comments

Figure S1 MRE11-dependent phosphorylation of Rad53 (PDF 144 kb)

Figure S2 Ddc2/Lcd1 and Ddc1 are phosphorylated in mre11Δ cells in the absence of DNA damaging agents.

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Grenon, M., Gilbert, C. & Lowndes, N. Checkpoint activation in response to double-strand breaks requires the Mre11/Rad50/Xrs2 complex. Nat Cell Biol 3, 844–847 (2001). https://doi.org/10.1038/ncb0901-844

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