A systematic screen identifies Saf5 as a link between splicing and transcription in fission yeast

Splicing is an important step of gene expression regulation in eukaryotes, as there are many mRNA precursors that can be alternatively spliced in different tissues, at different cell cycle phases or under different external stimuli. We have developed several integrated fluorescence-based in vivo spl...

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Autores: Borao Arriazu, Sonia, 1993-, Vega, Montserrat, Boronat i Llop, Susanna, 1965-, Hidalgo Hernando, Elena, Hümmer, Stefan, Ayté del Olmo, José
Tipo de recurso: artículo
Estado:Versión publicada
Fecha de publicación:2024
País:España
Institución:Varias* (Consorci de Biblioteques Universitáries de Catalunya, Centre de Serveis Científics i Acadèmics de Catalunya)
Repositorio:Recercat. Dipósit de la Recerca de Catalunya
OAI Identifier:oai:recercat.cat:10230/60777
Acceso en línea:http://hdl.handle.net/10230/60777
http://dx.doi.org/10.1371/journal.pgen.1011316
Access Level:acceso abierto
Palabra clave:Introns
Schizosaccharomyces pombe
DNA transcription
Gene regulation
Gene expression
Yellow fluorescent protein
RNA splicing
Saccharomyces cerevisiae
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spelling A systematic screen identifies Saf5 as a link between splicing and transcription in fission yeastBorao Arriazu, Sonia, 1993-Vega, MontserratBoronat i Llop, Susanna, 1965-Hidalgo Hernando, ElenaHümmer, StefanAyté del Olmo, JoséIntronsSchizosaccharomyces pombeDNA transcriptionGene regulationGene expressionYellow fluorescent proteinRNA splicingSaccharomyces cerevisiaeSplicing is an important step of gene expression regulation in eukaryotes, as there are many mRNA precursors that can be alternatively spliced in different tissues, at different cell cycle phases or under different external stimuli. We have developed several integrated fluorescence-based in vivo splicing reporter constructs that allow the quantification of fission yeast splicing in vivo on intact cells, and we have compared their splicing efficiency in a wild type strain and in a prp2-1 (U2AF65) genetic background, showing a clear dependency between Prp2 and a consensus signal at 5' splicing site (5'SS). To isolate novel genes involved in regulated splicing, we have crossed the reporter showing more intron retention with the Schizosaccharomyces pombe knock out collection. Among the candidate genes involved in the regulation of splicing, we have detected strong splicing defects in two of the mutants -Δcwf12, a member of the NineTeen Complex (NTC) and Δsaf5, a methylosome subunit that acts together with the survival motor neuron (SMN) complex in small nuclear ribonucleoproteins (snRNP) biogenesis. We have identified that strains with mutations in cwf12 have inefficient splicing, mainly when the 5'SS differs from the consensus. However, although Δsaf5 cells also have some dependency on 5'SS sequence, we noticed that when one intron of a given pre-mRNA was affected, the rest of the introns of the same pre-mRNA had high probabilities of being also affected. This observation points Saf5 as a link between transcription rate and splicing.This work was supported by grants BFU2018-PGC2018-097248-B-I00 and PID2022-136449NB-I00 funded by MICIU/AEI/10.13039/501100011033 and ERDF/EU to JA and by Unidad de Excelencia Maria de Maeztu (CEX2018-000792-M) to JA and EH. EH is a recipient of an ICREA Academia Award (Generalitat de Catalunya). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.Public Library of Science (PLoS)202420242024info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersionapplication/pdfapplication/pdfhttp://hdl.handle.net/10230/60777http://dx.doi.org/10.1371/journal.pgen.1011316reponame:Recercat. Dipósit de la Recerca de Catalunyainstname:Varias* (Consorci de Biblioteques Universitáries de Catalunya, Centre de Serveis Científics i Acadèmics de Catalunya)InglésPLoS Genet. 2024 Jun 4;20(6):e1011316info:eu-repo/grantAgreement/ES/2PE/PGC2018-097248-B-I00info:eu-repo/grantAgreement/ES/3PE/PID2022-136449NB-I00info:eu-repo/grantAgreement/ES/2PE/CEX2018-000792-M© 2024 Borao et al. This is an open access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.http://creativecommons.org/licenses/by/4.0/info:eu-repo/semantics/openAccessoai:recercat.cat:10230/607772026-05-29T05:05:01Z
dc.title.none.fl_str_mv A systematic screen identifies Saf5 as a link between splicing and transcription in fission yeast
title A systematic screen identifies Saf5 as a link between splicing and transcription in fission yeast
spellingShingle A systematic screen identifies Saf5 as a link between splicing and transcription in fission yeast
Borao Arriazu, Sonia, 1993-
Introns
Schizosaccharomyces pombe
DNA transcription
Gene regulation
Gene expression
Yellow fluorescent protein
RNA splicing
Saccharomyces cerevisiae
title_short A systematic screen identifies Saf5 as a link between splicing and transcription in fission yeast
title_full A systematic screen identifies Saf5 as a link between splicing and transcription in fission yeast
title_fullStr A systematic screen identifies Saf5 as a link between splicing and transcription in fission yeast
title_full_unstemmed A systematic screen identifies Saf5 as a link between splicing and transcription in fission yeast
title_sort A systematic screen identifies Saf5 as a link between splicing and transcription in fission yeast
dc.creator.none.fl_str_mv Borao Arriazu, Sonia, 1993-
Vega, Montserrat
Boronat i Llop, Susanna, 1965-
Hidalgo Hernando, Elena
Hümmer, Stefan
Ayté del Olmo, José
author Borao Arriazu, Sonia, 1993-
author_facet Borao Arriazu, Sonia, 1993-
Vega, Montserrat
Boronat i Llop, Susanna, 1965-
Hidalgo Hernando, Elena
Hümmer, Stefan
Ayté del Olmo, José
author_role author
author2 Vega, Montserrat
Boronat i Llop, Susanna, 1965-
Hidalgo Hernando, Elena
Hümmer, Stefan
Ayté del Olmo, José
author2_role author
author
author
author
author
dc.subject.none.fl_str_mv Introns
Schizosaccharomyces pombe
DNA transcription
Gene regulation
Gene expression
Yellow fluorescent protein
RNA splicing
Saccharomyces cerevisiae
topic Introns
Schizosaccharomyces pombe
DNA transcription
Gene regulation
Gene expression
Yellow fluorescent protein
RNA splicing
Saccharomyces cerevisiae
description Splicing is an important step of gene expression regulation in eukaryotes, as there are many mRNA precursors that can be alternatively spliced in different tissues, at different cell cycle phases or under different external stimuli. We have developed several integrated fluorescence-based in vivo splicing reporter constructs that allow the quantification of fission yeast splicing in vivo on intact cells, and we have compared their splicing efficiency in a wild type strain and in a prp2-1 (U2AF65) genetic background, showing a clear dependency between Prp2 and a consensus signal at 5' splicing site (5'SS). To isolate novel genes involved in regulated splicing, we have crossed the reporter showing more intron retention with the Schizosaccharomyces pombe knock out collection. Among the candidate genes involved in the regulation of splicing, we have detected strong splicing defects in two of the mutants -Δcwf12, a member of the NineTeen Complex (NTC) and Δsaf5, a methylosome subunit that acts together with the survival motor neuron (SMN) complex in small nuclear ribonucleoproteins (snRNP) biogenesis. We have identified that strains with mutations in cwf12 have inefficient splicing, mainly when the 5'SS differs from the consensus. However, although Δsaf5 cells also have some dependency on 5'SS sequence, we noticed that when one intron of a given pre-mRNA was affected, the rest of the introns of the same pre-mRNA had high probabilities of being also affected. This observation points Saf5 as a link between transcription rate and splicing.
publishDate 2024
dc.date.none.fl_str_mv 2024
2024
2024
dc.type.none.fl_str_mv info:eu-repo/semantics/article
info:eu-repo/semantics/publishedVersion
format article
status_str publishedVersion
dc.identifier.none.fl_str_mv http://hdl.handle.net/10230/60777
http://dx.doi.org/10.1371/journal.pgen.1011316
url http://hdl.handle.net/10230/60777
http://dx.doi.org/10.1371/journal.pgen.1011316
dc.language.none.fl_str_mv Inglés
language_invalid_str_mv Inglés
dc.relation.none.fl_str_mv PLoS Genet. 2024 Jun 4;20(6):e1011316
info:eu-repo/grantAgreement/ES/2PE/PGC2018-097248-B-I00
info:eu-repo/grantAgreement/ES/3PE/PID2022-136449NB-I00
info:eu-repo/grantAgreement/ES/2PE/CEX2018-000792-M
dc.rights.none.fl_str_mv http://creativecommons.org/licenses/by/4.0/
info:eu-repo/semantics/openAccess
rights_invalid_str_mv http://creativecommons.org/licenses/by/4.0/
eu_rights_str_mv openAccess
dc.format.none.fl_str_mv application/pdf
application/pdf
dc.publisher.none.fl_str_mv Public Library of Science (PLoS)
publisher.none.fl_str_mv Public Library of Science (PLoS)
dc.source.none.fl_str_mv reponame:Recercat. Dipósit de la Recerca de Catalunya
instname:Varias* (Consorci de Biblioteques Universitáries de Catalunya, Centre de Serveis Científics i Acadèmics de Catalunya)
instname_str Varias* (Consorci de Biblioteques Universitáries de Catalunya, Centre de Serveis Científics i Acadèmics de Catalunya)
reponame_str Recercat. Dipósit de la Recerca de Catalunya
collection Recercat. Dipósit de la Recerca de Catalunya
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repository.mail.fl_str_mv
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