Genome-wide Map of R-Loop-Induced Damage Reveals How a Subset of R-Loops Contributes to Genomic Instability

Chromatin immuno-precipitation (ChIP) of Rad52-bound to ssDNA that was compatible with next-generation sequencing to map sites of DNA damage induced by accumulation of genomic DNA-RNA hybrids. Examination of RAD52 signal in strains defective for DNA-RNA hybrid removal. Cells possess different DNA-RNA hybrid removal machineries like the RNases-H (RNH1 and RNH201 in yeast), DNA-RNA helicases (the essential Sen1 gene in yeast) and Topoisomerases (Top1 gene used in this study). We removed the activity of those enzymes and analyzed the pattern of Rad52 localization by Rad52-ChIPseq as a readout to DNA damage induced by DNA-RNA hybrids.