Studying Topoisomerase 1-Mediated Damage at Genomic Ribonucleotides

Abstract

Ribonucleotides incorporated into DNA by the DNA polymerases can be incised by Topoisomerase 1 (Top1) to initiate removal of ribonucleotides from the genome. This Top1-dependent ribonucleotide removal has been demonstrated to result in multiple forms of genome instability in yeast. Here, we describe both quantitative and qualitative assays to identify mutations and other forms of DNA damage resulting from Top1-cleavage at unrepaired genomic ribonucleotides.

Keywords: DNA replication; Genome instability; Mutagenesis; RNase H2; Replication stress; Ribonucleotide excision repair; Topoisomerase 1.