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An intronic RNA structure modulates expression of the mRNA biogenesis factor Sus1

Sus1 is a conserved protein involved in chromatin remodeling and mRNA biogenesis. Unlike most yeast genes, the SUS1 pre-mRNA of Saccharomyces cerevisiae contains two introns and is alternatively spliced, retaining one or both introns in response to changes in environmental conditions. SUS1 splicing...

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Detalles Bibliográficos
Autores principales: AbuQattam, Ali, Gallego, José, Rodríguez-Navarro, Susana
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Cold Spring Harbor Laboratory Press 2016
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4691836/
https://www.ncbi.nlm.nih.gov/pubmed/26546116
http://dx.doi.org/10.1261/rna.054049.115
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author AbuQattam, Ali
Gallego, José
Rodríguez-Navarro, Susana
author_facet AbuQattam, Ali
Gallego, José
Rodríguez-Navarro, Susana
author_sort AbuQattam, Ali
collection PubMed
description Sus1 is a conserved protein involved in chromatin remodeling and mRNA biogenesis. Unlike most yeast genes, the SUS1 pre-mRNA of Saccharomyces cerevisiae contains two introns and is alternatively spliced, retaining one or both introns in response to changes in environmental conditions. SUS1 splicing may allow the cell to control Sus1 expression, but the mechanisms that regulate this process remain unknown. Using in silico analyses together with NMR spectroscopy, gel electrophoresis, and UV thermal denaturation experiments, we show that the downstream intron (I2) of SUS1 forms a weakly stable, 37-nucleotide stem–loop structure containing the branch site near its apical loop and the 3′ splice site after the stem terminus. A cellular assay revealed that two of four mutants containing altered I2 structures had significantly impaired SUS1 expression. Semiquantitative RT-PCR experiments indicated that all mutants accumulated unspliced SUS1 pre-mRNA and/or induced distorted levels of fully spliced mRNA relative to wild type. Concomitantly, Sus1 cellular functions in histone H2B deubiquitination and mRNA export were affected in I2 hairpin mutants that inhibited splicing. This work demonstrates that I2 structure is relevant for SUS1 expression, and that this effect is likely exerted through modulation of splicing.
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spelling pubmed-46918362016-01-07 An intronic RNA structure modulates expression of the mRNA biogenesis factor Sus1 AbuQattam, Ali Gallego, José Rodríguez-Navarro, Susana RNA Article Sus1 is a conserved protein involved in chromatin remodeling and mRNA biogenesis. Unlike most yeast genes, the SUS1 pre-mRNA of Saccharomyces cerevisiae contains two introns and is alternatively spliced, retaining one or both introns in response to changes in environmental conditions. SUS1 splicing may allow the cell to control Sus1 expression, but the mechanisms that regulate this process remain unknown. Using in silico analyses together with NMR spectroscopy, gel electrophoresis, and UV thermal denaturation experiments, we show that the downstream intron (I2) of SUS1 forms a weakly stable, 37-nucleotide stem–loop structure containing the branch site near its apical loop and the 3′ splice site after the stem terminus. A cellular assay revealed that two of four mutants containing altered I2 structures had significantly impaired SUS1 expression. Semiquantitative RT-PCR experiments indicated that all mutants accumulated unspliced SUS1 pre-mRNA and/or induced distorted levels of fully spliced mRNA relative to wild type. Concomitantly, Sus1 cellular functions in histone H2B deubiquitination and mRNA export were affected in I2 hairpin mutants that inhibited splicing. This work demonstrates that I2 structure is relevant for SUS1 expression, and that this effect is likely exerted through modulation of splicing. Cold Spring Harbor Laboratory Press 2016-01 /pmc/articles/PMC4691836/ /pubmed/26546116 http://dx.doi.org/10.1261/rna.054049.115 Text en © 2015 AbuQattam et al.; Published by Cold Spring Harbor Laboratory Press for the RNA Society http://creativecommons.org/licenses/by/4.0/ This article, published in RNA, is available under a Creative Commons License (Attribution 4.0 International), as described at http://creativecommons.org/licenses/by/4.0/.
spellingShingle Article
AbuQattam, Ali
Gallego, José
Rodríguez-Navarro, Susana
An intronic RNA structure modulates expression of the mRNA biogenesis factor Sus1
title An intronic RNA structure modulates expression of the mRNA biogenesis factor Sus1
title_full An intronic RNA structure modulates expression of the mRNA biogenesis factor Sus1
title_fullStr An intronic RNA structure modulates expression of the mRNA biogenesis factor Sus1
title_full_unstemmed An intronic RNA structure modulates expression of the mRNA biogenesis factor Sus1
title_short An intronic RNA structure modulates expression of the mRNA biogenesis factor Sus1
title_sort intronic rna structure modulates expression of the mrna biogenesis factor sus1
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4691836/
https://www.ncbi.nlm.nih.gov/pubmed/26546116
http://dx.doi.org/10.1261/rna.054049.115
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