GSE124144
Rnase L reprograms translation by widespread mRNA turnover escaped by antiviral mRNAs
In response to foreign and endogenous double-stranded RNA (dsRNA), protein kinase R (PKR) and ribonuclease L (RNase L) reprogram translation in mammalian cells. PKR inhibits translation initiation through eIF2  phosphorylation, which triggers stress granule (SG) formation and promotes translation of stress responsive mRNAs. The mechanisms of RNase L-driven translation repression, its contribution to SG assembly, and its regulation of dsRNA stress-induced mRNAs are unknown. We demonstrate that RNase L drives translational shut-off in response to dsRNA by promoting widespread turnover of mRNAs. This alters stress granule assembly and reprograms translation by only allowing for the translation of mRNAs resistant to RNase L degradation, including numerous antiviral mRNAs such as IFN- . Individual cells differentially activate dsRNA responses revealing variation that can affect cellular outcomes. This identifies bulk mRNA degradation and the resistance of antiviral mRNAs as the mechanism by which RNaseL reprograms translation in response to dsRNA. Overall design: Total RNA from parental (WT) and RNase L knockout (RLKO) A549 cells before and after 6-hours post-transfection of poly(I:C) (500-ng/ml) was isolated for library preparation. Three independent samples from each cell line and condition were analyzed. ERRC spike-in controls were added prior to rRNA depletion and library preparation.