Concerted 2-5A-Mediated mRNA Decay and Transcription Reprogram Protein Synthesis in dsRNA Response

Sneha Rath; Eliza Prangley; Jesse Donovan; Kaitlin Demarest; Yigal Meir; Ned Wingreen; Alexei Korennykh

To test whether a uniform mRNA decay is taking place, we used qPCR that we designed to detect full-length mRNAs (Fig. 4A ). Using this assay we observed a rapid decay of a several abundant basal mRNAs (Fig. 4B ). As expected, the decay was absent in RNase L -/cells (Fig. S4A ). All tested mRNAs were cleaved rapidly. The decay traces for all mRNAs leveled before 100% cleavage, suggesting that cells have 2-5AMD-sensitive and 2-5AMD-resistant mRNA pools. The size of the resistant fraction was higher for PRKDC and SON mRNAs (Fig. 4B , dotted lines). By quantifying loglinear regions of the data (Fig. 4B , inset graphs), we determined first-order decay . CC-BY 4.0 International license is made available under a The copyright holder for this preprint (which was not peer-reviewed) is the author/funder. It . https://doi.org/10.1101/484675 doi: bioRxiv preprint kinetics for each mRNA (Fig. 4C ). The housekeeping mRNAs decay considerably faster than rRNA, and on the same time scale as the translational arrest ( Fig. 3D vs 4C ).