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SF3B1/Hsh155 HEAT motif mutations affect interaction with the spliceosomal ATPase Prp5, resulting in altered branch site selectivity in pre-mRNA splicing

Mutations in the U2 snRNP component SF3B1 are prominent in myelodysplastic syndromes (MDSs) and other cancers and have been shown recently to alter branch site (BS) or 3′ splice site selection in splicing. However, the molecular mechanism of altered splicing is not known. We show here that hsh155 mu...

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Autores principales: Tang, Qing, Rodriguez-Santiago, Susana, Wang, Jing, Pu, Jia, Yuste, Andrea, Gupta, Varun, Moldón, Alberto, Xu, Yong-Zhen, Query, Charles C.
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/PMC5238730/
https://www.ncbi.nlm.nih.gov/pubmed/28087715
http://dx.doi.org/10.1101/gad.291872.116
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author Tang, Qing
Rodriguez-Santiago, Susana
Wang, Jing
Pu, Jia
Yuste, Andrea
Gupta, Varun
Moldón, Alberto
Xu, Yong-Zhen
Query, Charles C.
author_facet Tang, Qing
Rodriguez-Santiago, Susana
Wang, Jing
Pu, Jia
Yuste, Andrea
Gupta, Varun
Moldón, Alberto
Xu, Yong-Zhen
Query, Charles C.
author_sort Tang, Qing
collection PubMed
description Mutations in the U2 snRNP component SF3B1 are prominent in myelodysplastic syndromes (MDSs) and other cancers and have been shown recently to alter branch site (BS) or 3′ splice site selection in splicing. However, the molecular mechanism of altered splicing is not known. We show here that hsh155 mutant alleles in Saccharomyces cerevisiae, counterparts of SF3B1 mutations frequently found in cancers, specifically change splicing of suboptimal BS pre-mRNA substrates. We found that Hsh155p interacts directly with Prp5p, the first ATPase that acts during spliceosome assembly, and localized the interacting regions to HEAT (Huntingtin, EF3, PP2A, and TOR1) motifs in SF3B1 associated with disease mutations. Furthermore, we show that mutations in these motifs from both human disease and yeast genetic screens alter the physical interaction with Prp5p, alter branch region specification, and phenocopy mutations in Prp5p. These and other data demonstrate that mutations in Hsh155p and Prp5p alter splicing because they change the direct physical interaction between Hsh155p and Prp5p. This altered physical interaction results in altered loading (i.e., “fidelity”) of the BS–U2 duplex into the SF3B complex during prespliceosome formation. These results provide a mechanistic framework to explain the consequences of intron recognition and splicing of SF3B1 mutations found in disease.
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spelling pubmed-52387302017-06-15 SF3B1/Hsh155 HEAT motif mutations affect interaction with the spliceosomal ATPase Prp5, resulting in altered branch site selectivity in pre-mRNA splicing Tang, Qing Rodriguez-Santiago, Susana Wang, Jing Pu, Jia Yuste, Andrea Gupta, Varun Moldón, Alberto Xu, Yong-Zhen Query, Charles C. Genes Dev Research Paper Mutations in the U2 snRNP component SF3B1 are prominent in myelodysplastic syndromes (MDSs) and other cancers and have been shown recently to alter branch site (BS) or 3′ splice site selection in splicing. However, the molecular mechanism of altered splicing is not known. We show here that hsh155 mutant alleles in Saccharomyces cerevisiae, counterparts of SF3B1 mutations frequently found in cancers, specifically change splicing of suboptimal BS pre-mRNA substrates. We found that Hsh155p interacts directly with Prp5p, the first ATPase that acts during spliceosome assembly, and localized the interacting regions to HEAT (Huntingtin, EF3, PP2A, and TOR1) motifs in SF3B1 associated with disease mutations. Furthermore, we show that mutations in these motifs from both human disease and yeast genetic screens alter the physical interaction with Prp5p, alter branch region specification, and phenocopy mutations in Prp5p. These and other data demonstrate that mutations in Hsh155p and Prp5p alter splicing because they change the direct physical interaction between Hsh155p and Prp5p. This altered physical interaction results in altered loading (i.e., “fidelity”) of the BS–U2 duplex into the SF3B complex during prespliceosome formation. These results provide a mechanistic framework to explain the consequences of intron recognition and splicing of SF3B1 mutations found in disease. Cold Spring Harbor Laboratory Press 2016-12-15 /pmc/articles/PMC5238730/ /pubmed/28087715 http://dx.doi.org/10.1101/gad.291872.116 Text en © 2016 Tang et al.; Published by Cold Spring Harbor Laboratory Press http://creativecommons.org/licenses/by-nc/4.0/ This article is distributed exclusively by Cold Spring Harbor Laboratory Press for the first six months after the full-issue publication date (see http://genesdev.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/.
spellingShingle Research Paper
Tang, Qing
Rodriguez-Santiago, Susana
Wang, Jing
Pu, Jia
Yuste, Andrea
Gupta, Varun
Moldón, Alberto
Xu, Yong-Zhen
Query, Charles C.
SF3B1/Hsh155 HEAT motif mutations affect interaction with the spliceosomal ATPase Prp5, resulting in altered branch site selectivity in pre-mRNA splicing
title SF3B1/Hsh155 HEAT motif mutations affect interaction with the spliceosomal ATPase Prp5, resulting in altered branch site selectivity in pre-mRNA splicing
title_full SF3B1/Hsh155 HEAT motif mutations affect interaction with the spliceosomal ATPase Prp5, resulting in altered branch site selectivity in pre-mRNA splicing
title_fullStr SF3B1/Hsh155 HEAT motif mutations affect interaction with the spliceosomal ATPase Prp5, resulting in altered branch site selectivity in pre-mRNA splicing
title_full_unstemmed SF3B1/Hsh155 HEAT motif mutations affect interaction with the spliceosomal ATPase Prp5, resulting in altered branch site selectivity in pre-mRNA splicing
title_short SF3B1/Hsh155 HEAT motif mutations affect interaction with the spliceosomal ATPase Prp5, resulting in altered branch site selectivity in pre-mRNA splicing
title_sort sf3b1/hsh155 heat motif mutations affect interaction with the spliceosomal atpase prp5, resulting in altered branch site selectivity in pre-mrna splicing
topic Research Paper
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5238730/
https://www.ncbi.nlm.nih.gov/pubmed/28087715
http://dx.doi.org/10.1101/gad.291872.116
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