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The carboxy terminus of NBS1 is required for induction of apoptosis by the MRE11 complex

Abstract

The MRE11 complex (MRE11, RAD50 and NBS1) and the ataxia-telangiectasia mutated (ATM) kinase function in the same DNA damage response pathway to effect cell cycle checkpoint activation and apoptosis1,2,3. The functional interaction between the MRE11 complex and ATM has been proposed to require a conserved C-terminal domain of NBS1 for recruitment of ATM to sites of DNA damage4,5. Human Nijmegen breakage syndrome (NBS) cells and those derived from multiple mouse models of NBS express a hypomorphic NBS1 allele that exhibits impaired ATM activity despite having an intact C-terminal domain3,6,7,8,9,10,11. This indicates that the NBS1 C terminus is not sufficient for ATM function. We derived Nbs1ΔC/ΔC mice in which the C-terminal ATM interaction domain is deleted. Nbs1ΔC/ΔC cells exhibit intra-S-phase checkpoint defects, but are otherwise indistinguishable from wild-type cells with respect to other checkpoint functions, ionizing radiation sensitivity and chromosome stability. However, multiple tissues of Nbs1ΔC/ΔC mice showed a severe apoptotic defect, comparable to that of ATM- or CHK2-deficient animals. Analysis of p53 transcriptional targets and ATM substrates showed that, in contrast to the phenotype of Chk2-/- mice, NBS1ΔC does not impair the induction of proapoptotic genes. Instead, the defects observed in Nbs1ΔC/ΔC result from impaired phosphorylation of ATM targets including SMC1 and the proapoptotic factor, BID.

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Figure 1: Generation of Nbs1 ΔC/ΔC mice.
Figure 2: Cellular phenotypes of Nbs1ΔC/ΔC.
Figure 3: Apoptotic phenotypes of Nbs1ΔC/ΔC.
Figure 4: Apoptotic signalling in Nbs1ΔC/ΔC.

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References

  1. Stracker, T. H., Theunissen, J. W., Morales, M. & Petrini, J. H. The Mre11 complex and the metabolism of chromosome breaks: the importance of communicating and holding things together. DNA Repair 3, 845–854 (2004)

    Article  CAS  Google Scholar 

  2. Morales, M. et al. The Rad50S allele promotes ATM-dependent DNA damage responses and suppresses ATM deficiency: implications for the Mre11 complex as a DNA damage sensor. Genes Dev. 19, 3043–3054 (2005)

    Article  CAS  Google Scholar 

  3. Shiloh, Y. ATM and related protein kinases: safeguarding genome integrity. Nature Rev. Cancer 3, 155–168 (2003)

    Article  CAS  Google Scholar 

  4. You, Z., Chahwan, C., Bailis, J., Hunter, T. & Russell, P. ATM activation and its recruitment to damaged DNA require binding to the C terminus of Nbs1. Mol. Cell. Biol. 25, 5363–5379 (2005)

    Article  CAS  Google Scholar 

  5. Falck, J., Coates, J. & Jackson, S. P. Conserved modes of recruitment of ATM, ATR and DNA-PKcs to sites of DNA damage. Nature 434, 605–611 (2005)

    Article  ADS  CAS  Google Scholar 

  6. Carney, J. P. et al. The hMre11/hRad50 protein complex and Nijmegen breakage syndrome: linkage of double-strand break repair to the cellular DNA damage response. Cell 93, 477–486 (1998)

    Article  CAS  Google Scholar 

  7. Maser, R. S., Zinkel, R. & Petrini, J. H. J. An alternative mode of translation permits production of a variant NBS1 protein from the common Nijmegen breakage syndrome allele. Nature Genet. 27, 417–421 (2001)

    Article  CAS  Google Scholar 

  8. Maser, R. S. et al. The MRE11 complex and DNA replication: linkage to E2F and sites of DNA synthesis. Mol. Cell. Biol. 21, 6006–6016 (2001)

    Article  CAS  Google Scholar 

  9. Williams, B. R. et al. A murine model of Nijmegen breakage syndrome. Curr. Biol. 12, 648–653 (2002)

    Article  CAS  Google Scholar 

  10. Kang, J., Bronson, R. T. & Xu, Y. Targeted disruption of NBS1 reveals its roles in mouse development and DNA repair. EMBO J. 21, 1447–1455 (2002)

    Article  CAS  Google Scholar 

  11. Difilippantonio, S. et al. Role of Nbs1 in the activation of the Atm kinase revealed in humanized mouse models. Nature Cell Biol. 7, 675–685 (2005)

    Article  CAS  Google Scholar 

  12. Barlow, C. et al. Atm-deficient mice: a paradigm of ataxia telangiectasia. Cell 86, 159–171 (1996)

    Article  CAS  Google Scholar 

  13. Xu, Y. & Baltimore, D. Dual roles of ATM in the cellular response to radiation and in cell growth control. Genes Dev. 10, 2401–2410 (1996)

    Article  CAS  Google Scholar 

  14. Xu, Y. et al. Targeted disruption of ATM leads to growth retardation, chromosomal fragmentation during meiosis, immune defects, and thymic lymphoma. Genes Dev. 10, 2411–2422 (1996)

    Article  CAS  Google Scholar 

  15. Yamazaki, V., Wegner, R. D. & Kirchgessner, C. U. Characterization of cell cycle checkpoint responses after ionizing radiation in Nijmegen breakage syndrome cells. Cancer Res. 58, 2316–2322 (1998)

    CAS  PubMed  Google Scholar 

  16. Kang, J. et al. Functional interaction of H2AX, NBS1, and p53 in ATM-dependent DNA damage responses and tumor suppression. Mol. Cell. Biol. 25, 661–670 (2005)

    Article  CAS  Google Scholar 

  17. Kang, J., Bronson, R. & Xu, Y. Targeted disruption of NBS1 reveals its roles in mouse development and DNA repair. EMBO J. 21, 1447–1455 (2002)

    Article  CAS  Google Scholar 

  18. Kitagawa, R., Bakkenist, C. J., McKinnon, P. J. & Kastan, M. B. Phosphorylation of SMC1 is a critical downstream event in the ATM–NBS1–BRCA1 pathway. Genes Dev. 18, 1423–1438 (2004)

    Article  CAS  Google Scholar 

  19. Bakkenist, C. J. & Kastan, M. B. DNA damage activates ATM through intermolecular autophosphorylation and dimer dissociation. Nature 421, 499–506 (2003)

    Article  ADS  CAS  Google Scholar 

  20. Bender, C. F. et al. Cancer predisposition and hematopoietic failure in Rad50S/S mice. Genes Dev. 16, 2237–2251 (2002)

    Article  CAS  Google Scholar 

  21. Takai, H. et al. Chk2-deficient mice exhibit radioresistance and defective p53-mediated transcription. EMBO J. 21, 5195–5205 (2002)

    Article  CAS  Google Scholar 

  22. Hirao, A. et al. Chk2 is a tumor suppressor that regulates apoptosis in both an ataxia telangiectasia mutated (ATM)-dependent and an ATM-independent manner. Mol. Cell. Biol. 22, 6521–6532 (2002)

    Article  CAS  Google Scholar 

  23. Falck, J., Petrini, J. H., Williams, B. R., Lukas, J. & Bartek, J. The DNA damage-dependent intra–S phase checkpoint is regulated by parallel pathways. Nature Genet. 30, 290–294 (2002)

    Article  Google Scholar 

  24. Kamer, I. et al. Proapoptotic BID is an ATM effector in the DNA-damage response. Cell 122, 593–603 (2005)

    Article  CAS  Google Scholar 

  25. Zinkel, S. S. et al. A role for proapoptotic BID in the DNA-damage response. Cell 122, 579–591 (2005)

    Article  CAS  Google Scholar 

  26. Kastan, M. B. et al. A mammalian cell cycle checkpoint pathway utilizing p53 and GADD45 is defective in ataxia-telangiectasia. Cell 71, 587–597 (1992)

    Article  CAS  Google Scholar 

  27. Lee, J. H. & Paull, T. T. ATM activation by DNA double-strand breaks through the Mre11–Rad50–Nbs1 complex. Science 308, 551–554 (2005)

    Article  ADS  CAS  Google Scholar 

  28. Theunissen, J. W. & Petrini, J. H. Methods for studying the cellular response to DNA damage: influence of the Mre11 complex on chromosome metabolism. Methods Enzymol. 409, 251–284 (2006)

    Article  CAS  Google Scholar 

  29. Theunissen, J. W. et al. Checkpoint failure and chromosomal instability without lymphomagenesis in Mre11ATLD1/ATLD1 mice. Mol. Cell 12, 1511–1523 (2003)

    Article  CAS  Google Scholar 

  30. Liu, P., Jenkins, N. A. & Copeland, N. G. A highly efficient recombineering-based method for generating conditional knockout mutations. Genome Res. 13, 476–484 (2003)

    Article  CAS  Google Scholar 

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Acknowledgements

We thank N. Copeland, N. Jenkins, and C. Adelman for assistance with recombineering and ES cell culture, J. Theunissen for assistance with checkpoint and apoptotic analysis, G. Oltz and E. Rhuley for AC1 ES cells, Y. Shiloh for anti-ATM (MAT3) antibodies, and Petrini laboratory members for helpful suggestions. T.H.S. was supported by an NRSA fellowship and this work was supported by NIH grants awarded to J.H.P. and the Joel and Joan Smilow Initiative.

Author Contributions T.H.S. and J.H.P. conceived the experiments and wrote the paper. T.H.S., M.M., S.S.C., and H.H. performed the experiments.

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Correspondence to John H. J. Petrini.

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Stracker, T., Morales, M., Couto, S. et al. The carboxy terminus of NBS1 is required for induction of apoptosis by the MRE11 complex. Nature 447, 218–221 (2007). https://doi.org/10.1038/nature05740

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