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Autophosphorylation at serine 1987 is dispensable for murine Atm activation in vivo

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.

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Figure 1: Atm null and heterozygous mice express Atm-S1987A from a BAC transgene.
Figure 2: Expression of Atm-S1987A reconstitutes lymphocyte development and restores genomic stability.
Figure 3: S1987 phosphorylation is dispensable for Atm kinase activity and recruitment to DNA breaks.

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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.).

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Correspondence to André Nussenzweig.

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This file contains Supplementary Figures 1–9 and Supplementary Methods. (PDF 11940 kb)

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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

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