Global analysis of RNA oxidation in Saccharomyces cerevisiae.
Oxidative RNA damage has been linked to loss of RNA function and to the development of many human age-related diseases. Consequently, a need exists for methods to identify and quantify the extent of RNA oxidation on a genome-wide basis. We developed such a method by combining affinity selection of mRNA containing 8-hydroxyguanine with high throughput DNA sequencing. We demonstrate that this assay is suitable for detecting differences in the extent of oxidation between RNA transcripts. We applied this method to the yeast Saccharomyces cerevisiae grown under physiological conditions and in response to hydrogen peroxide, and detected significantly oxidized RNA transcripts.
Department of Genome Sciences, University of Washington, Seattle, WA 98195, USA.
Anastasia
A
McKinlay
Wayne
W
Gerard
Stanley
S
Fields
D057166:High-Throughput Screening Assays / Q000379:methods*; D010084:Oxidation-Reduction; D012331:RNA, Fungal / Q000235:genetics / Q000378:metabolism*; D012441:Saccharomyces cerevisiae / Q000235:genetics; D029701:Saccharomyces cerevisiae Proteins / Q000235:genetics / Q000378:metabolism*
D012331:RNA, Fungal; D029701:Saccharomyces cerevisiae Proteins