研究组蛋白去乙酰化酶在神经元DNA双链断裂修复中的作用。

Examining the Role of HDACs in DNA Double-Strand Break Repair in Neurons.

作者信息

Pao Ping-Chieh, Penney Jay, Tsai Li-Huei

机构信息

Picower Institute for Learning and Memory, Massachusetts Institute of Technology, Cambridge, MA, USA.

Department of Brain and Cognitive Sciences, Massachusetts Institute of Technology, Cambridge, MA, USA.

出版信息

Methods Mol Biol. 2019;1983:225-234. doi: 10.1007/978-1-4939-9434-2_13.

DOI:10.1007/978-1-4939-9434-2_13

PMID:31087301

Abstract

Histone deacetylases (HDACs) modulate chromatin structure by removing acetyl groups from histones. Upon DNA double-strand breaks (DSBs), deacetylation of H3K56 and H4K16 by HDACs occurs immediately at break sites, and is crucial for DSB repair. Here we describe two assays that examine defective DSB repair caused by HDAC inhibition in primary cortical neurons: single-cell gel electrophoresis to assay DNA integrity (the comet assay) and western blot analysis for γH2AX, a phosphorylated histone variant associated with DSBs.

摘要

组蛋白去乙酰化酶（HDACs）通过去除组蛋白上的乙酰基团来调节染色质结构。在DNA双链断裂（DSBs）时，HDACs会立即在断裂位点使H3K56和H4K16去乙酰化，这对DSB修复至关重要。在此，我们描述了两种检测原代皮质神经元中由HDAC抑制引起的DSB修复缺陷的方法：用于检测DNA完整性的单细胞凝胶电泳（彗星试验）以及针对γH2AX的蛋白质印迹分析，γH2AX是一种与DSBs相关的磷酸化组蛋白变体。