2026-04-17T20:42:53Zhttps://recercat.cat/oai/request

oai:recercat.cat:10230/607772025-12-20T16:48:21Zcom_2072_6col_2072_452952

00925njm 22002777a 4500 dc Borao Arriazu, Sonia, 1993- author Vega, Montserrat author Boronat i Llop, Susanna, 1965- author Hidalgo Hernando, Elena author Hümmer, Stefan author Ayté del Olmo, José author 2024-07-18T06:56:29Z 2024-07-18T06:56:29Z 2024 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. 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. Introns Schizosaccharomyces pombe DNA transcription Gene regulation Gene expression Yellow fluorescent protein RNA splicing Saccharomyces cerevisiae A systematic screen identifies Saf5 as a link between splicing and transcription in fission yeast