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Role of Nbs1 in the activation of the Atm kinase revealed in humanized mouse models

Nature Cell Biology volume 7, pages 675–685 (2005)Cite this article

Abstract

Nijmegen breakage syndrome (NBS) is a chromosomal fragility disorder that shares clinical and cellular features with ataxia telangiectasia. Here we demonstrate that Nbs1-null B cells are defective in the activation of ataxia-telangiectasia-mutated (Atm) in response to ionizing radiation, whereas ataxia-telangiectasia- and Rad3-related (Atr)-dependent signalling and Atm activation in response to ultraviolet light, inhibitors of DNA replication, or hypotonic stress are intact. Expression of the main human NBS allele rescues the lethality of Nbs1−/− mice, but leads to immunodeficiency, cancer predisposition, a defect in meiotic progression in females and cell-cycle checkpoint defects that are associated with a partial reduction in Atm activity. The Mre11 interaction domain of Nbs1 is essential for viability, whereas the Forkhead-associated (FHA) domain is required for T-cell and oocyte development and efficient DNA damage signalling. Reconstitution of Nbs1 knockout mice with various mutant isoforms demonstrates the biological impact of impaired Nbs1 function at the cellular and organismal level.

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Figure 1: Human Nbs1 functionally replaces mouse Nbs1.
Figure 2: Infertility in hNbs1657Δ5 female mice.
Figure 3: Abnormal T-cell development in hNbs1657Δ5 mice.
Figure 4: Class-switch recombination and lymphoma predisposition in hNbs1657Δ5 mice.
Figure 5: Response to cellular stress in Nbs1 knockout and hypomorphic mutant B cells.
Figure 6: Association of Mre11 with DSBs in hNbs1657Δ5 B cells.
Figure 7: Inefficient accumulation of Mre11 at DSBs and at sites of DNA replication.
Figure 8: Role of the Nbs1 FHA and Mre11-binding domains in vivo.

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Acknowledgements

We thank J. Petrini, S. Ganesan and A. Lee for generously providing antibodies, R. Kitagawa for experimental protocols, A. Hohenstein and N. Puri for assistance with western blotting, L. Stapleton and T. Ried for chromosome paints, M. Kastan and Y. Shiloh for helpful suggestions, and M. Difilippantonio and M. Pellegrini for comments on the manuscript. This work was supported in part by grants from NIH and the Leukemia Society to M.C.N. M.C.N. is an HHMI investigator.

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Author notes

  1. Oscar Fernandez-Capetillo

    Present address: Genomic Instability Group, Molecular Oncology Program, Spanish National Cancer Center (CNIO), Madrid, 28029, Spain

Authors and Affiliations

  1. Experimental Immunology Branch, National Cancer Institute, National Institutes of Health, Bethesda, 20892, MD, USA

    Simone Difilippantonio, Arkady Celeste, Oscar Fernandez-Capetillo, Hua-Tang Chen, Francois Van Laethem, Michael Kruhlak & André Nussenzweig

  2. Laboratory of Molecular Immunology, The Rockefeller University, New York, 10021, NY, USA

    Bernardo Reina San Martin & Michel Nussenzweig

  3. Howard Hughes Medical Institute, The Rockefeller University, New York, 10021, NY, USA

    Michel Nussenzweig

  4. Mouse Cancer Genetics Program, National Cancer Institute at Frederick, 1050 Boyles Street, Frederick, 21702, MD, USA

    Yong-Ping Yang & Shyam Sharan

  5. Genetics and Biochemistry Branch, National Institutes of Diabetes and Digestive and Kidney Diseases, National Institutes of Health, Bethesda, 20892, MD, USA

    Galina V. Petukhova & R. Daniel Camerini-Otero

  6. Division of Veterinary Resources, Office of Research Services, Office of the Director, National Institutes of Health, Bethesda, 20892, MD, USA

    Michael Eckhaus

  7. SAIC-Frederick, National Cancer Institute-Frederick Cancer Research and Development Center, Frederick, 21702, MD, USA

    Lionel Feigenbaum

  8. Molecular Cytology Core Facility and Molecular Biology Program, Memorial Sloan-Kettering Cancer Center, New York, 10021, NY, USA

    Katia Manova

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Correspondence to André Nussenzweig.

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Difilippantonio, S., Celeste, A., Fernandez-Capetillo, O. et al. Role of Nbs1 in the activation of the Atm kinase revealed in humanized mouse models. Nat Cell Biol 7, 675–685 (2005). https://doi.org/10.1038/ncb1270

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