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

Methods Mol Biol. 2019; 
Song C, Grummt I.
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摘要

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.

关键词

DDX21; DNA-RNA hybrids; Gel electrophoresis; Helicase; Immunopurification; Protein acetylation; R-loops; Recombinant proteins; Unwinding of DNA-RNA structures