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DNA damage activates ATM through intermolecular autophosphorylation and dimer dissociation

Abstract

The ATM protein kinase, mutations of which are associated with the human disease ataxia–telangiectasia, mediates responses to ionizing radiation in mammalian cells. Here we show that ATM is held inactive in unirradiated cells as a dimer or higher-order multimer, with the kinase domain bound to a region surrounding serine 1981 that is contained within the previously described ‘FAT’ domain. Cellular irradiation induces rapid intermolecular autophosphorylation of serine 1981 that causes dimer dissociation and initiates cellular ATM kinase activity. Most ATM molecules in the cell are rapidly phosphorylated on this site after doses of radiation as low as 0.5 Gy, and binding of a phosphospecific antibody is detectable after the introduction of only a few DNA double-strand breaks in the cell. Activation of the ATM kinase seems to be an initiating event in cellular responses to irradiation, and our data indicate that ATM activation is not dependent on direct binding to DNA strand breaks, but may result from changes in the structure of chromatin.

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Figure 1: Metabolic labelling of ATM in response to ionizing radiation.
Figure 2: Intermolecular autophosphorylation of ATM serine 1981 occurs in vivo after irradiation.
Figure 3: S1981A-ATM has kinase activity but is dominant-inhibitory in cells.
Figure 4: Interaction of ATM domains with proteins.
Figure 5: Kinetics, stoichiometry and detection sensitivity of ATM phosphorylation.
Figure 6: Activation of ATM by chromatin-active agents that do not cause DNA breaks.

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Acknowledgements

We acknowledge the technical assistance of H. Davis, M. Reis and D. Woods. We thank S. Powell, Y. Shiloh and R. Abraham for providing reagents, and all members of the Kastan laboratory and J. Cleveland, R. Ivey, T. Izard, P. McKinnon, D. Coffey and C. Sherr for reading the manuscript or for helpful discussions. This work was supported by grants from the National Institutes of Health and by the American Lebanese Syrian Associated Charities of the St Jude Children's Research Hospital.

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Correspondence to Michael B. Kastan.

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Bakkenist, C., Kastan, M. DNA damage activates ATM through intermolecular autophosphorylation and dimer dissociation. Nature 421, 499–506 (2003). https://doi.org/10.1038/nature01368

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