Activation of the antiviral factor RNase L triggers translation of non-coding mRNA sequences

Ribonuclease L (RNase L) is activated as part of the innate immune response and plays an important role in the clearance of viral infections. When activated, it endonucleolytically cleaves both viral and host RNAs, leading to a global reduction in protein synthesis. However, it remains unknown how widespread RNA decay, and consequent changes in the translatome, promote the elimination of viruses. To study how this altered transcriptome is translated, we assayed the global distribution of ribosomes in RNase L activated human cells with ribosome profiling. We found that RNase L activation leads to a substantial increase in the fraction of translating ribosomes in ORFs internal to coding sequences (iORFs) and ORFs within 59 and 39 UTRs (uORFs and dORFs). Translation of these alternative ORFs was dependent on RNase L9s cleavage activity, suggesting that mRNA decay fragments are translated to produce short peptides that may be important for antiviral activity.