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研究解旋酶乙酰化在调节R环稳定性中作用的实验方法

Experimental Approaches to Investigate the Role of Helicase Acetylation in Regulating R-Loop Stability.

作者信息

Song Chenlin, Grummt Ingrid

机构信息

Division of Molecular Biology of the Cell II, German Cancer Research Center (DKFZ), DKFZ-ZMBH Alliance, Heidelberg, Germany.

出版信息

Methods Mol Biol. 2019;1983:237-253. doi: 10.1007/978-1-4939-9434-2_14.

Abstract

R-loops are three-stranded nucleic acid structures composed of a DNA-RNA heteroduplex and a displaced single-stranded DNA. Although R-loops serve important roles in transcription and chromatin structure, they are also a major threat to genome stability. Cells prevent accumulation of genomic R-loops by mechanisms that remove these structures, such as ribonucleases which digest DNA-RNA hybrids and helicases which unwind R-loops. Here we describe methods to monitor resolvement of R-loops by the helicase DDX21 focussing on the impact of acetylation on helicase activity.

摘要

R环是由DNA-RNA异源双链体和一条单链DNA组成的三链核酸结构。尽管R环在转录和染色质结构中发挥着重要作用,但它们也是基因组稳定性的重大威胁。细胞通过去除这些结构的机制来防止基因组R环的积累,比如消化DNA-RNA杂交体的核糖核酸酶和解开R环的解旋酶。在这里,我们描述了通过解旋酶DDX21监测R环分解的方法,重点关注乙酰化对解旋酶活性的影响。

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