Roychoudhury Shrabasti, Pramanik Suravi, Harris Hannah L, Bhakat Kishor K
Department of Genetics, Cell Biology and Anatomy, University of Nebraska Medical Center, Omaha, NE, USA.
Fred and Pamela Buffet Cancer Center, University of Nebraska Medical Center, Omaha, NE, USA.
Methods Mol Biol. 2019;1983:191-206. doi: 10.1007/978-1-4939-9434-2_11.
Protein posttranslational modifications (PTMs), including acetylation, have emerged as important regulators for controlling many cellular processes. DNA base excision repair (BER), a highly coordinated multistep cellular process, is primarily involved in the repair of both endogenous and drug-induced exogenous DNA base damages. BER relies on sequential recruitment and coordinated actions of multiple proteins. Increasing evidence suggests that acetylation of lysine residues of BER proteins facilitates fine-tuning of enzymatic activities, protein-protein interactions, and coordination of the steps in BER pathway. In this chapter, we describe detailed in vitro and in vivo approaches to examine the effect of acetylation on BER enzymes, focusing on the impact of acetylation of AP-endonuclease (APE1), a key enzyme in BER pathway, on its DNA damage repair activity, substrate-binding, and subcellular localization.
蛋白质翻译后修饰（PTM），包括乙酰化，已成为控制许多细胞过程的重要调节因子。DNA碱基切除修复（BER）是一个高度协调的多步骤细胞过程，主要参与内源性和药物诱导的外源性DNA碱基损伤的修复。BER依赖于多种蛋白质的顺序募集和协同作用。越来越多的证据表明，BER蛋白赖氨酸残基的乙酰化有助于对酶活性、蛋白质-蛋白质相互作用以及BER途径中各步骤的协调进行微调。在本章中，我们详细描述了体外和体内方法，以研究乙酰化对BER酶的影响，重点关注BER途径中的关键酶——脱嘌呤嘧啶内切酶（APE1）的乙酰化对其DNA损伤修复活性、底物结合和亚细胞定位的影响。
