Sun Yingli, Du Fengxia
Cancer and Epigenetic Group, Key Laboratory of Genomic and Precision Medicine, China Gastrointestinal Cancer Research Center, Beijing Institute of Genomics, Chinese Academy of Sciences, No.1 Beichen West Road, Chaoyang District, Beijing, 100101, China.
Methods Mol Biol. 2017;1599:157-162. doi: 10.1007/978-1-4939-6955-5_12.
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.
ATM的激活在DNA双链断裂修复途径中至关重要。Tip60组蛋白乙酰转移酶（HAT）对ATM的乙酰化在响应DNA损伤时激活ATM激酶活性中起关键作用。ATM通过其FATC结构域与Tip60形成稳定复合物。Tip60使ATM的赖氨酸3016乙酰化，这种乙酰化诱导ATM的激活。Tip60对ATM乙酰化的研究包括多种技术，如体外激酶测定、系统诱变、蛋白质免疫印迹。在此，我们描述如何使用乙酰化特异性抗体研究ATM的乙酰化。
