Identification of ATM-interacting proteins by co-immunoprecipitation and glutathione-s-transferase (GST) pull-down assays

Abstract

The ATM kinase is a master regulator of the DNA damage response, and can interact with more than 700 proteins in response to DNA damage. These interactions play a critical role in fine-tuning the response of ATM to multiple cellular stressors, and can play both a positive or negative role in regulating its activity. Here, we detail using protein-protein interaction methods, including co-immunoprecipitation and Glutathione-S-transferase (GST) fusion protein pull-down assays to understand the molecular interactions of ATM. These assays give valuable functional insights into the role of ATM, as they are easy to establish within the laboratory, are not overly laborious, and are easily reproducible.