Global RNA stability profiling reveals stabilization of P-body-enriched transcripts in the absence of human Dcp2

Multiple RNA decapping enzymes coexist in mammalian cells to regulate decay of distinct subsets of cellular transcripts, but their specificity remains incompletely defined to date. Here we present a global profile of RNA stability changes in human Dcp2 knockout cells via TimeLapse-seq. We demonstrate that P-body enrichment is the strongest correlate of Dcp2-dependent RNA decay, and that post-transcriptional modifications such as m6A present additive effect for Dcp2 targeting. Importantly, our data support a model in which P-bodies are sites that sort translationally repressed transcripts for cytoplasmic decay through additional molecular marks. Overall design: RNA isolated from cells +/- s4U feed was subjected to oxidative-nucleophilic-aromatic-substitution chemistry and sequenced. All sequencing for experimental samples was performed in duplicate per condition assessed, while negative controls were performed as single replicates without s4U treatment.