Integrative transcriptomic analysis suggests new autoregulatory splicing events coupled with nonsense-mediated mRNA decay

Nucleic Acids Res. 2019 Jun 4;47(10):5293-5306. doi: 10.1093/nar/gkz193.

Abstract

Nonsense-mediated decay (NMD) is a eukaryotic mRNA surveillance system that selectively degrades transcripts with premature termination codons (PTC). Many RNA-binding proteins (RBP) regulate their expression levels by a negative feedback loop, in which RBP binds its own pre-mRNA and causes alternative splicing to introduce a PTC. We present a bioinformatic analysis integrating three data sources, eCLIP assays for a large RBP panel, shRNA inactivation of NMD pathway, and shRNA-depletion of RBPs followed by RNA-seq, to identify novel such autoregulatory feedback loops. We show that RBPs frequently bind their own pre-mRNAs, their exons respond prominently to NMD pathway disruption, and that the responding exons are enriched with nearby eCLIP peaks. We confirm previously proposed models of autoregulation in SRSF7 and U2AF1 genes and present two novel models, in which (i) SFPQ binds its mRNA and promotes switching to an alternative distal 3'-UTR that is targeted by NMD, and (ii) RPS3 binding activates a poison 5'-splice site in its pre-mRNA that leads to a frame shift and degradation by NMD. We also suggest specific splicing events that could be implicated in autoregulatory feedback loops in RBM39, HNRNPM, and U2AF2 genes. The results are available through a UCSC Genome Browser track hub.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • 3' Untranslated Regions
  • Alternative Splicing
  • Codon, Nonsense*
  • Computational Biology
  • Exons
  • Frameshift Mutation
  • Heterogeneous-Nuclear Ribonucleoprotein Group M / metabolism
  • Humans
  • Nonsense Mediated mRNA Decay*
  • Nuclear Proteins / metabolism
  • RNA Precursors / metabolism
  • RNA Splicing*
  • RNA, Messenger / metabolism
  • RNA, Small Interfering / metabolism*
  • RNA-Binding Proteins / metabolism
  • Serine-Arginine Splicing Factors / metabolism
  • Spliceosomes
  • Splicing Factor U2AF / metabolism
  • Transcriptome*

Substances

  • 3' Untranslated Regions
  • Codon, Nonsense
  • HCC1 autoantigen
  • HNRNPM protein, human
  • Heterogeneous-Nuclear Ribonucleoprotein Group M
  • Nuclear Proteins
  • RNA Precursors
  • RNA, Messenger
  • RNA, Small Interfering
  • RNA-Binding Proteins
  • SRSF7 protein, human
  • Splicing Factor U2AF
  • U2AF1 protein, human
  • U2AF2 protein, human
  • Serine-Arginine Splicing Factors