Functional Characterization of ATM Kinase Using Acetylation-Specific Antibodies

Methods Mol Biol. 2017:1599:157-162. doi: 10.1007/978-1-4939-6955-5_12.

Abstract

The activation of ATM is critical in the DNA double strand breaks repair pathway. Acetylation of ATM by Tip60 histone acetyltransferase (HAT) plays a key role in the activation of ATM kinase activity in response to DNA damage. ATM forms a stable complex with Tip60 through the FATC domain of ATM. Tip60 acetylates lysine3016 of ATM, and this acetylation induces the activation of ATM. Several techniques are included in the study of ATM acetylation by Tip60, such as in vitro kinase assay, systematic mutagenesis, western blots. Here, we describe how to study the acetylation of ATM using acetylation-specific antibodies.

Keywords: ATM acetylation; ATM activation; Acetylation; Tip60; Western blot.

MeSH terms

  • Acetylation
  • Ataxia Telangiectasia Mutated Proteins / genetics
  • Ataxia Telangiectasia Mutated Proteins / metabolism*
  • Blotting, Western
  • DNA Damage / genetics
  • DNA Damage / physiology
  • Humans
  • Lysine Acetyltransferase 5 / genetics
  • Lysine Acetyltransferase 5 / metabolism

Substances

  • KAT5 protein, human
  • Lysine Acetyltransferase 5
  • Ataxia Telangiectasia Mutated Proteins