Post-transcriptional control of mRNAs by RNA-binding proteins (RBPs) has a prominent role in the regulation of gene expression. RPBs interact with mRNAs to control their biogenesis, splicing, transport, localization, translation, and stability. Defects in such regulation can lead to a wide range of human diseases from neurological disorders to cancer.
Many RBPs are conserved between Caenorhabditis elegans and humans, and several RBPs are known to regulate apoptosis in the adult C. elegans germline. How these RBPs control apoptosis is, however, largely unknown. We identified wago-4, one of the 27 worm argonautes, in an RNAi screen as novel regulator of germ cell apoptosis. In an in vivo CLIP experiment we found that wago-4 mRNA is controlled by MINA-1, another novel apoptosis relevant RBP.
By RNAi experiments we determined that WAGO-4 acts specifically in the germ line. This is supported by the exclusive expression of WAGO-4 in the germline ranging from the distal tip cell to the region of late oocyte formation. Also, the identification of protein-protein interaction partners led to the conclusion that WAGO-4 plays a role in RNAi.
A system wide analysis including quantitative proteome data (SILAC) and transcriptome data (RNAseq) of a wago-4 deletion mutant compared to wild type pointed at a post-transcriptional regulatory function of WAGO-4 during apoptosis. Our comprehensive approach allowed us to build a model of the germ cell apoptosis RNA regulon and to broaden our understanding of how RBPs orchestrate different cellular events.
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