Ribosome profiling at isoform level reveals evolutionary conserved impacts of differential splicing on the proteome

• Published in: Nature communications 2020-04, Vol.11 (1), p.1768-1768, Article 1768
• Main Authors: Reixachs-Solé, Marina, Ruiz-Orera, Jorge, Albà, M. Mar, Eyras, Eduardo
• Format: Article
• Language: English
• Subjects: 38/91; 631/114; 631/337/1645/1946; 631/337/2019; 631/337/574/1789; 631/45/500; Alternative splicing; Alternative Splicing - genetics; Animals; Computational Biology - methods; Evolutionary conservation; Exons; Genes; Glioma; Humanities and Social Sciences; Humans; Isoforms; Mice; multidisciplinary; Open reading frames; Pipelines; Protein Isoforms - genetics; Protein Isoforms - metabolism; Proteome - genetics; Proteome - metabolism; Proteomes; Ribosomes - genetics; Ribosomes - metabolism; RNA Splicing - genetics; RNA Splicing - physiology; Science; Science (multidisciplinary); Splicing; Transcription; Transcriptome - genetics; Translation
• ISSN: 2041-1723; 2041-1723
• DOI: 10.1038/s41467-020-15634-w

The differential production of transcript isoforms from gene loci is a key cellular mechanism. Yet, its impact in protein production remains an open question. Here, we describe ORQAS (ORF quantification pipeline for alternative splicing), a pipeline for the translation quantification of individual transcript isoforms using ribosome-protected mRNA fragments (ribosome profiling). We find evidence of translation for 40–50% of the expressed isoforms in human and mouse, with 53% of the expressed genes having more than one translated isoform in human, and 33% in mouse. Differential splicing analysis revealed that about 40% of the splicing changes at RNA level are concordant with changes in translation. Furthermore, orthologous cassette exons between human and mouse preserve the directionality of the change, and are enriched in microexons in a comparison between glia and glioma. ORQAS leverages ribosome profiling to uncover a widespread and evolutionarily conserved impact of differential splicing on translation, particularly of microexon-containing isoforms. Genes express multiple mRNA isoforms through alternative processing. Here the authors analyze ribosome profiling data with ORQAS (ORF quantification pipeline for alternative splicing) and report that 40–50% of the expressed mRNA isoforms are translated.