The surveillance of pre-mRNA splicing is an early step in C. elegans RNAi of endogenous genes

• Published in: Genes & development 2018-05, Vol.32 (9-10), p.670-681
• Main Authors: Newman, Martin A, Ji, Fei, Fischer, Sylvia E J, Anselmo, Anthony, Sadreyev, Ruslan I, Ruvkun, Gary
• Format: Article
• Language: English
• Subjects: Animals; Caenorhabditis elegans - genetics; Caenorhabditis elegans Proteins - genetics; Gene Expression Regulation - genetics; Introns - genetics; Mutation; Research Paper; RNA Interference - physiology; RNA Precursors - metabolism; RNA Splicing; RNA Splicing Factors - genetics; RNA, Small Nuclear - genetics; Spliceosomes - metabolism
• ISSN: 0890-9369; 1549-5477
• DOI: 10.1101/gad.311514.118

RNAi pathways detect and silence foreign nucleic acids such as viruses as well as endogenous genes in many species. The phylogenetic profile across eukaryotes of proteins that mediate key steps in RNAi is correlated with the profiles of multiple mRNA splicing proteins and with intron number, suggesting that RNAi may surveil mRNA splicing to detect the divergent or absent introns of viruses. Here we examine the role of mRNA splicing in RNAi We found that viable null mutations in U1 and U2 small nuclear ribonucleic protein (snRNP)-specific splicing factor genes cause defects in RNAi. The U1A ortholog is required for normal ERGO-1 Argonaute class 26G siRNA biogenesis, -splicing of the transcript, and targeting of poorly conserved gene transcripts by WAGO Argonaute class 22G siRNAs. We found that gene transcripts engaged by the siRNA-generating machinery are poorly conserved, possess few introns, and often have introns that are divergent from introns with strong consensus splicing sites found in highly conserved genes. We present biochemical evidence that RNAi targeted transcripts are tightly bound to spliceosomes. These findings suggest multiple layers of regulation by the spliceosome at early steps of small RNA-mediated gene silencing.