Ribosome clearance during RNA interference

• DOI: 10.1261/rna.070813.119
• Publication Date: 2019-05-20T21:05:49Z
• Authors: Makena N. Pule; Marissa L. Glover; Andrew Z. Fire; Joshua A. Arribere
• Keywords: 0301 basic medicine; 571; Messenger; nonstop; Article; 03 medical and health sciences; Genetics; Helminth; Animals; RNA, Messenger; Caenorhabditis elegans; Caenorhabditis elegans Proteins; Sequence Analysis, RNA; PELOTA; SKI; Endonucleases; ribosome; RNAi; C. elegans; RNA; RNA Interference; Generic health relevance; Biochemistry and Cell Biology; RNA, Helminth; Sequence Analysis; Ribosomes; Biotechnology; Developmental Biology

In the course of identifying and cleaving RNA, the RNAi machinery must encounter and contend with the megadalton-sized ribosomes that carry out translation. We investigated this interface by examining the fate of actively translated mRNAs subjected to RNAi in C. elegans. Quantifying RNA levels (RNA-seq) and ongoing translation (Ribo-seq), we found there is a greater fold repression of ongoing translation than expected from loss of RNA alone, observing stronger translation repression relative to RNA repression for multiple, independent double-stranded RNA triggers, and for multiple genes. In animals that lack the RNA helicase SKI complex and the ribosome rescue factor PELOTA, ribosomes stall on the 3′ edges of mRNAs at and upstream of the RNAi trigger. One model to explain these observations is that ribosomes are actively cleared from mRNAs by SKI and PELO during or following mRNA cleavage. Our results expand prior studies that show a role for the SKI RNA helicase complex in removing RNA targets following RNAi in flies and plants, illuminating the widespread role of the nonstop translation surveillance in RNA silencing during RNAi. Our results are also consistent with proposals that RNAi can attack messages during active translation.