Analysis of NRAS RNA G-quadruplex binding proteins reveals DDX3X as a novel interactor of cellular G-quadruplex containing transcripts.
Nucleic acids research
Oxford University Press
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Herdy, B., Mayer, C., Varshney, D., Marsico, G., Murat, P., Taylor, C., D'Santos, C., et al. (2018). Analysis of NRAS RNA G-quadruplex binding proteins reveals DDX3X as a novel interactor of cellular G-quadruplex containing transcripts.. Nucleic acids research, 46 (21), 11592-11604. https://doi.org/10.1093/nar/gky861
RNA G-quadruplexes (rG4s) are secondary structures in mRNAs known to influence RNA post-transcriptional mechanisms thereby impacting neurodegenerative disease and cancer. A detailed knowledge of rG4-protein interactions is vital to enable the understanding of rG4 function. Herein, we describe a systematic, unbiased affinity proteomics approach that identified 80 high-confidence interactors that assemble on the rG4 located in the 5’untranslated region (UTR) of the NRAS oncogene. Novel rG4 interactors included DDX3X, DDX5, DDX17, GRSF1 and NSUN5. The majority of identified proteins contained a glycine-arginine (GAR) domain and notably GAR-domain mutation in DDX3X and DDX17 abrogated rG4 binding. Identification of DDX3X targets by transcriptome-wide individual-nucleotide resolution UV-crosslinking and affinity enrichment (iCLAE) revealed a striking association with 5’UTR rG4-containing transcripts which was reduced upon GAR-domain mutation. Our work highlights hitherto unrecognized features of rG4 structure-protein interactions that highlight new roles of rG4 structures in mRNA post-transcriptional control.
Hela Cells, Cytoplasm, Humans, RNA, Messenger, 5' Untranslated Regions, Reproducibility of Results, Genes, ras, DEAD-box RNA Helicases, G-Quadruplexes, Protein Domains
Cancer Research UK (CB4330)
European Research Council (339778)
European Commission Horizon 2020 (H2020) Marie Sk?odowska-Curie actions (702476)
External DOI: https://doi.org/10.1093/nar/gky861
This record's URL: https://www.repository.cam.ac.uk/handle/1810/285369
Attribution 4.0 International
Licence URL: https://creativecommons.org/licenses/by/4.0/