Full-length transcript alterations in human bronchial epithelial cells with U2AF1 S34F mutations

Life Sci Alliance. 2023 Jul 24;6(10):e202000641. doi: 10.26508/lsa.202000641. Print 2023 Oct.

Abstract

U2AF1 is one of the most recurrently mutated splicing factors in lung adenocarcinoma and has been shown to cause transcriptome-wide pre-mRNA splicing alterations; however, the full-length altered mRNA isoforms associated with the mutation are largely unknown. To better understand the impact U2AF1 has on full-length isoform fate and function, we conducted high-throughput long-read cDNA sequencing from isogenic human bronchial epithelial cells with and without a U2AF1 S34F mutation. We identified 49,366 multi-exon transcript isoforms, more than half of which did not match GENCODE or short-read-assembled isoforms. We found 198 transcript isoforms with significant expression and usage changes relative to WT, only 68% of which were assembled by short reads. Expression of isoforms from immune-related genes is largely down-regulated in mutant cells and without observed splicing changes. Finally, we reveal that isoforms likely targeted by nonsense-mediated decay are down-regulated in U2AF1 S34F cells, suggesting that isoform changes may alter the translational output of those affected genes. Altogether, our work provides a resource of full-length isoforms associated with U2AF1 S34F in lung cells.

Publication types

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

MeSH terms

  • Epithelial Cells* / metabolism
  • Humans
  • Mutation / genetics
  • Protein Isoforms / genetics
  • Protein Isoforms / metabolism
  • RNA Splicing* / genetics
  • Splicing Factor U2AF / genetics
  • Splicing Factor U2AF / metabolism

Substances

  • Splicing Factor U2AF
  • Protein Isoforms
  • U2AF1 protein, human