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Methods Enzymol. 2006;408:529-39.

Purification and biochemical characterization of ataxia-telangiectasia mutated and Mre11/Rad50/Nbs1.

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  • 1Institute of Cellular and Molecular Biology, University of Texas at Austin, USA.


Ataxia-telangiectasia mutated (ATM) is a serine-threonine kinase that is activated by DNA double strand breaks to phosphorylate many cellular proteins involved in cell cycle regulation and DNA repair. We have shown previously that the activation of ATM can be reconstituted in an in vitro system using recombinant human ATM. In this system, ATM activity is dependent on the Mre11/Rad50/Nbs1 (MRN) complex and linear DNA, similar to requirements observed in human cells. This chapter describes methods used for the overexpression and purification of human ATM and MRN, as well as a protocol for in vitro kinase assays.

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