Native RNA sequencing in fission yeast reveals frequent alternative splicing isoforms

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• dc.contributor.author Montañés, José Carlos
• dc.contributor.author Huertas, Marta
• dc.contributor.author Moro, Simone G.
• dc.contributor.author Blevins, William Robert, 1987-
• dc.contributor.author Carmona, Mercè
• dc.contributor.author Ayté del Olmo, José
• dc.contributor.author Hidalgo Hernando, Elena
• dc.contributor.author Albà Soler, Mar
• dc.date.accessioned 2023-04-24T06:15:38Z
• dc.date.available 2023-04-24T06:15:38Z
• dc.date.issued 2022
• dc.description.abstract The unicellular yeast Schizosaccharomyces pombe (fission yeast) retains many of the splicing features observed in humans and is thus an excellent model to study the basic mechanisms of splicing. Nearly half the genes contain introns, but the impact of alternative splicing in gene regulation and proteome diversification remains largely unexplored. Here we leverage Oxford Nanopore Technologies native RNA sequencing (dRNA), as well as ribosome profiling data, to uncover the full range of polyadenylated transcripts and translated open reading frames. We identify 332 alternative isoforms affecting the coding sequences of 262 different genes, 97 of which occur at frequencies higher than 20%, indicating that functional alternative splicing in S. pombe is more prevalent than previously suspected. Intron retention events make about 80% of the cases; these events may be involved in the regulation of gene expression and, in some cases, generate novel protein isoforms, as supported by ribosome profiling data in 18 of the intron retention isoforms. One example is the rpl22 gene, in which intron retention is associated with the translation of a protein of only 13 amino acids. We also find that lowly expressed transcripts tend to have longer poly(A) tails than highly expressed transcripts, highlighting an interdependence between poly(A) tail length and transcript expression level. Finally, we discover 214 novel transcripts that are not annotated, including 158 antisense transcripts, some of which also show translation evidence. The methodologies described in this work open new opportunities to study the regulation of splicing in a simple eukaryotic model.
• dc.description.sponsorship This work benefited from preliminary Nanopore RNA-seq data analyses performed by Bea Calvo and Audald Lloret-Villas, as well as discussions with Eduardo Eyras and Ivan de la Rubia. We acknowledge funding from Ministerio de Ciencia e Innovación (MCI), Agencia Estatal de Investigación (AEI) grant PGC2018–094091-B-I00, cofunded by Fondo Europeo de Desarrollo Regional (FEDER), and from Agència de Gestió d'Ajuts Universitaris i de Recerca (AGAUR), Generalitat de Catalunya, grant 2017SGR01020.
• dc.format.mimetype application/pdf
• dc.identifier.citation Montañés JC, Huertas M, Moro SG, Blevins WR, Carmona M, Ayté J, Hidalgo E, Albà MM. Native RNA sequencing in fission yeast reveals frequent alternative splicing isoforms. Genome Res. 2022 May 26;32(6):1215-27. DOI: 10.1101/gr.276516.121
• dc.identifier.doi http://dx.doi.org/10.1101/gr.276516.121
• dc.identifier.issn 1088-9051
• dc.identifier.uri http://hdl.handle.net/10230/56544
• dc.language.iso eng
• dc.publisher Cold Spring Harbor Laboratory Press (CSHL Press)
• dc.relation.ispartof Genome Res. 2022 May 26;32(6):1215-27
• dc.relation.projectID info:eu-repo/grantAgreement/ES/2PE/PGC2018–094091-B-I00
• dc.rights © 2022 Montañés et al.; Published by Cold Spring Harbor Laboratory Press. This article is distributed exclusively by Cold Spring Harbor Laboratory Press for the first six months after the full-issue publication date (see https://genome.cshlp.org/site/misc/terms.xhtml). After six months, it is available under a Creative Commons License (Attribution-NonCommercial 4.0 International), as described at http://creativecommons.org/licenses/by-nc/4.0/.
• dc.rights.accessRights info:eu-repo/semantics/openAccess
• dc.rights.uri http://creativecommons.org/licenses/by-nc/4.0/
• dc.subject.other Regulació genètica
• dc.subject.other Seqüència d'aminoàcids
• dc.title Native RNA sequencing in fission yeast reveals frequent alternative splicing isoforms
• dc.type info:eu-repo/semantics/article
• dc.type.version info:eu-repo/semantics/publishedVersion