Ribosome recycling factor ABCE1 depletion inhibits nonsense-mediated mRNA decay by promoting stop codon readthrough

Abstract

Nonsense-mediated mRNA decay (NMD) is an essential post-transcriptional surveillance pathway in vertebrates that appears to be mechanistically linked with translation termination. To gain more insight into this connection, we interfered with translation termination by depleting human cells of the ribosome recycling factor ABCE1, which resulted in an upregulation of many but not all endogenous NMD-sensitive mRNAs. Notably, the suppression of NMD on these mRNAs occurs at a step prior to their SMG6-mediated endonucleolytic cleavage. Ribosome profiling revealed that ABCE1 depletion results in ribosome stalling at stop codons and increased ribosome occupancy in 3 UTRs, indicative of enhanced stop codon readthrough or re-initiation. Using reporter genes, we further demonstrate that the absence of ABCE1 indeed increases the rate of readthrough, which would explain the observed NMD inhibition, since enhanced readthrough has been previously shown to render NMD-sensitive transcripts resistant to NMD by displacing NMD triggering factors like UPF1 and exon junction complexes (EJCs) from the 3 UTR. Collectively, our results show that improper ribosome disassembly interferes with proper NMD activation. HighlightsO_LIABCE1 knockdown suppresses NMD of many NMD-sensitive mRNAs C_LIO_LIThe observed NMD inhibition occurs at a stage prior to SMG6-mediated cleavage of the mRNA C_LIO_LIABCE1 depletion enhances ribosome occupancy at stop codons and in the 3 UTR C_LIO_LIABCE1 depletion enhances readthrough of the stop codon C_LIO_LIEnhanced readthrough inhibits NMD, presumably by clearing the 3 UTR of NMD factors C_LI