Abstract
The ATM (ataxia telangiectasia mutated) protein kinase is activated under physiological and pathological conditions that induce DNA double-strand breaks (DSBs). Loss of ATM or failure of its activation in humans and mice lead to defective cellular responses to DSBs, such as cell cycle checkpoints, radiation sensitivity, immune dysfunction, infertility and cancer predisposition. A widely used biological marker to identify the active form of ATM is the autophosphorylation of ATM at a single, conserved serine residue (Ser 1981 in humans; Ser 1987 in mouse)1. Here we show that Atm-dependent responses are functional at the organismal and cellular level in mice that express a mutant form of Atm (mutation of Ser to Ala at position 1987) as their sole Atm species. Moreover, the mutant protein does not exhibit dominant-negative interfering activity when expressed physiologically or overexpressed in the context of Atm heterozygous mice. These results suggest an alternative mode for stimulation of Atm by DSBs in which Atm autophosphorylation at Ser 1987, like trans-phosphorylation of downstream substrates, is a consequence rather than a cause of Atm activation.
This is a preview of subscription content, access via your institution
Access options
Subscribe to this journal
Receive 51 print issues and online access
$199.00 per year
only $3.90 per issue
Buy this article
- Purchase on Springer Link
- Instant access to full article PDF
Prices may be subject to local taxes which are calculated during checkout
Similar content being viewed by others
References
Bakkenist, C. J. & Kastan, M. B. DNA damage activates ATM through intermolecular autophosphorylation and dimer dissociation. Nature 421, 499–506 (2003)
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)
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)
Yang, Y. & Sharan, S. K. A simple two-step, ‘hit and fix’ method to generate subtle mutations in BACs using short denatured PCR fragments. Nucleic Acids Res. 31, e80 (2003)
Bartkova, J. et al. ATM activation in normal human tissues and testicular cancer. Cell Cycle 4, 838–845 (2005)
Ramiro, A. R. et al. Role of genomic instability and p53 in AID-induced c-myc–Igh translocations. Nature 440, 105–109 (2006)
Liu, Q. et al. Chk1 is an essential kinase that is regulated by Atr and required for the G(2)/M DNA damage checkpoint. Genes Dev. 14, 1448–1459 (2000)
Zachos, G., Rainey, M. D. & Gillespie, D. A. Chk1-deficient tumour cells are viable but exhibit multiple checkpoint and survival defects. EMBO J. 22, 713–723 (2003)
Takai, H. et al. Aberrant cell cycle checkpoint function and early embryonic death in Chk1-/- mice. Genes Dev. 14, 1439–1447 (2000)
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)
Bartek, J., Lukas, C. & Lukas, J. Checking on DNA damage in S phase. Nature Rev. Mol. Cell Biol. 5, 792–804 (2004)
Hickson, I. et al. Identification and characterization of a novel and specific inhibitor of the ataxia-telangiectasia mutated kinase ATM. Cancer Res. 64, 9152–9159 (2004)
Kozlov, S., Gueven, N., Keating, K., Ramsay, J. & Lavin, M. F. ATP activates ataxia-telangiectasia mutated (ATM) in vitro. Importance of autophosphorylation. J. Biol. Chem. 278, 9309–9317 (2003)
Uziel, T. et al. Requirement of the MRN complex for ATM activation by DNA damage. EMBO J. 22, 5612–5621 (2003)
Carson, C. T. et al. The Mre11 complex is required for ATM activation and the G2/M checkpoint. EMBO J. 22, 6610–6620 (2003)
Horejsi, Z. et al. Distinct functional domains of Nbs1 modulate the timing and magnitude of ATM activation after low doses of ionizing radiation. Oncogene 23, 3122–3127 (2004)
Cerosaletti, K. & Concannon, P. Independent roles for nibrin and Mre11-Rad50 in the activation and function of Atm. J. Biol. Chem. 279, 38813–38819 (2004)
Costanzo, V., Paull, T., Gottesman, M. & Gautier, J. Mre11 assembles linear DNA fragments into DNA damage signaling complexes. PLoS Biol. 2, E110 (2004)
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)
Mochan, T. A., Venere, M., DiTullio, R. A. Jr. & Halazonetis, T. D. 53BP1 and NFBD1/MDC1-Nbs1 function in parallel interacting pathways activating ataxia-telangiectasia mutated (ATM) in response to DNA damage. Cancer Res. 63, 8586–8591 (2003)
Sun, Y., Jiang, X., Chen, S., Fernandes, N. & Price, B. D. A role for the Tip60 histone acetyltransferase in the acetylation and activation of ATM. Proc. Natl Acad. Sci. USA 102, 13182–13187 (2005)
Gupta, A. et al. Involvement of human MOF in ATM function. Mol. Cell. Biol. 25, 5292–5305 (2005)
Ali, A. et al. Requirement of protein phosphatase 5 in DNA-damage-induced ATM activation. Genes Dev. 18, 249–254 (2004)
Lee, J. H. & Paull, T. T. ATM activation by DNA double-strand breaks through the Mre11-Rad50-Nbs1 complex. Science 308, 551–554 (2005)
Dupre, A., Boyer-Chatenet, L. & Gautier, J. Two-step activation of ATM by DNA and the Mre11–Rad50–Nbs1 complex. Nature Struct. Mol. Biol. 13, 451–457 (2006)
Goldstine, J. V. et al. Constitutive phosphorylation of ATM in lymphoblastoid cell lines from patients with ICF syndrome without downstream kinase activity. DNA Repair (Amst.) 5, 432–443 (2006)
Goodarzi, A. A. & Lees-Miller, S. P. Biochemical characterization of the ataxia-telangiectasia mutated (ATM) protein from human cells. DNA Repair (Amst.) 3, 753–767 (2004)
Cerosaletti, K., Wright, J. & Concannon, P. Active role for nibrin in the kinetics of atm activation. Mol. Cell. Biol. 26, 1691–1699 (2006)
Kruhlak, M. J. et al. Changes in chromatin structure and mobility in living cells at sites of DNA double-strand breaks. J. Cell Biol. 172, 823–834 (2006)
Andegeko, Y. et al. Nuclear retention of ATM at sites of DNA double strand breaks. J. Biol. Chem. 276, 38224–38230 (2001)
Acknowledgements
We thank S. Sharan for assistance with BAC recombineering; S. Jay, D. Winkler, F. Van Laethem, M. Kruhlak and M. Eckhhaus for technical assistance; G. Smith for providing ATM and DNA-PKcs small molecule inhibitors; T. Paull for providing human monomeric ATM; and O. Fernandez-Capetillo and A. Singer for helpful suggestions on the manuscript. This work was supported by the Intramural research Program of the NIH, National Cancer Institute, Center for Cancer Research, National Institute of Aging, and the AT Children's Project (grant to A.N.).
Author information
Authors and Affiliations
Corresponding author
Ethics declarations
Competing interests
Reprints and permissions information is available at npg.nature.com/reprintsandpermissions. The authors declare no competing financial interests.
Supplementary information
Supplementary Notes
This file contains Supplementary Figures 1–9 and Supplementary Methods. (PDF 11940 kb)
Rights and permissions
About this article
Cite this article
Pellegrini, M., Celeste, A., Difilippantonio, S. et al. Autophosphorylation at serine 1987 is dispensable for murine Atm activation in vivo. Nature 443, 222–225 (2006). https://doi.org/10.1038/nature05112
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1038/nature05112
This article is cited by
-
Bone Marrow Failure and Immunodeficiency Associated with Human RAD50 Variants
Journal of Clinical Immunology (2023)
-
Elevated inflammatory responses and targeted therapeutic intervention in a preclinical mouse model of ataxia-telangiectasia lung disease
Scientific Reports (2021)
-
ATM, ATR and DNA-PKcs kinases—the lessons from the mouse models: inhibition ≠ deletion
Cell & Bioscience (2020)
-
ATM, DNA-PKcs and ATR: shaping development through the regulation of the DNA damage responses
Genome Instability & Disease (2020)
-
Regulation of DNA damage-induced ATM activation by histone modifications
Genome Instability & Disease (2020)
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.