Poster Presentation 50 Years Shine-Dalgarno Symposium 2023

RNA Binding Motif Protein 47 (RBM47) C>U editing sites in the gut epithelium (#128)

David T Humphreys 1 , Madeleine Demuth 2 , Violeta Rayon 3 , Karin Tourle 2 , Tennille Sibbritt 2 , Jonathan Arthur 2 , Thomas Preiss 4 , Nina Papavasiliou 3 , Patrick P.L. Tam 2 , Nicolas Fossat 2
  1. Victor Chang Cardiac Research Instititute, Darlinghurst, NSW, Australia
  2. Children’s Medical Research Institute, Sydney, NSW, Australia
  3. Immune Diversity, Deutsches Krebsforschungszentrum, Heidelberg, Baden-Wurttemberg, Germany
  4. The John Curtin School of Medical Research, The Australian National University, Canberra, ACT, Australia

RBM47 is a RNA-binding protein that is found in many organs including the intestine, the liver, the lung, the pancreas and the kidney where it is mainly expressed in epithelial cells. We previously demonstrated that RBM47 is critical for Cytidine (C) to Uridine (U) RNA editing mediated by the deaminase APOBEC1 in the gut epithelium. In this study we undertook RNA-Seq analysis of the epithelium of the small intestine of wild type, Rbm47 deficient and Apobec knockout mice, to identify more than 1600 high confidence C>U editing events, a significant increase over previous collated ~250 C>U sites. We validated 537 selected sites using amplicon high throughput sequencing by comparing the signal from RNA and genomic DNA. These C>U editing sites were predominantly localised in the 3’UTR with the remainder being spread across coding, intronic and 5’UTRs. Across the 3’UTR, C>U editing was present in both highly conserved and non-conserved sites but not enriched in miRNA, polyA motif, or RNA-binding protein sites. Furthermore, editing events did not correlate with differential expression of the targeted genes. Exonic C>U editing sites were predominantly synonymous, but some non-synonymous changes, such as novel stop codon, do present.We confirmed the veracity of local sequence and RNA secondary structural requirements for RNA editing. Lastly, we also demonstrated the utility of the C>U editing data resource in identifying intestinal cell types that express edited transcripts through the re-analysis of single cell RNA-seq.