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The DExD/H-box ATPase Prp2p destabilizes and proofreads the catalytic RNA core of the spliceosome

After undergoing massive RNA and protein rearrangements during assembly, the spliceosome undergoes a final, more subtle, ATP-dependent rearrangement that is essential for catalysis. This rearrangement requires the DEAH-box protein Prp2p, an RNA-dependent ATPase. Prp2p has been implicated in destabil...

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Detalles Bibliográficos
Autores principales: Wlodaver, Alissa M., Staley, Jonathan P.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Cold Spring Harbor Laboratory Press 2014
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3923124/
https://www.ncbi.nlm.nih.gov/pubmed/24442613
http://dx.doi.org/10.1261/rna.042598.113
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author Wlodaver, Alissa M.
Staley, Jonathan P.
author_facet Wlodaver, Alissa M.
Staley, Jonathan P.
author_sort Wlodaver, Alissa M.
collection PubMed
description After undergoing massive RNA and protein rearrangements during assembly, the spliceosome undergoes a final, more subtle, ATP-dependent rearrangement that is essential for catalysis. This rearrangement requires the DEAH-box protein Prp2p, an RNA-dependent ATPase. Prp2p has been implicated in destabilizing interactions between the spliceosome and the protein complexes SF3 and RES, but a role for Prp2p in destabilizing RNA–RNA interactions has not been explored. Using directed molecular genetics in budding yeast, we have found that a cold-sensitive prp2 mutation is suppressed not only by mutations in SF3 and RES components but also by a range of mutations that disrupt the spliceosomal catalytic core element U2/U6 helix I, which is implicated in juxtaposing the 5′ splice site and branch site and in positioning metal ions for catalysis within the context of a putative catalytic triplex; indeed, mutations in this putative catalytic triplex also suppressed a prp2 mutation. Remarkably, we also found that prp2 mutations rescue lethal mutations in U2/U6 helix I. These data provide evidence that RNA elements that comprise the catalytic core are already formed at the Prp2p stage and that Prp2p destabilizes these elements, directly or indirectly, both to proofread spliceosome activation and to promote reconfiguration of the spliceosome to a fully competent, catalytic conformation.
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spelling pubmed-39231242015-03-01 The DExD/H-box ATPase Prp2p destabilizes and proofreads the catalytic RNA core of the spliceosome Wlodaver, Alissa M. Staley, Jonathan P. RNA Articles After undergoing massive RNA and protein rearrangements during assembly, the spliceosome undergoes a final, more subtle, ATP-dependent rearrangement that is essential for catalysis. This rearrangement requires the DEAH-box protein Prp2p, an RNA-dependent ATPase. Prp2p has been implicated in destabilizing interactions between the spliceosome and the protein complexes SF3 and RES, but a role for Prp2p in destabilizing RNA–RNA interactions has not been explored. Using directed molecular genetics in budding yeast, we have found that a cold-sensitive prp2 mutation is suppressed not only by mutations in SF3 and RES components but also by a range of mutations that disrupt the spliceosomal catalytic core element U2/U6 helix I, which is implicated in juxtaposing the 5′ splice site and branch site and in positioning metal ions for catalysis within the context of a putative catalytic triplex; indeed, mutations in this putative catalytic triplex also suppressed a prp2 mutation. Remarkably, we also found that prp2 mutations rescue lethal mutations in U2/U6 helix I. These data provide evidence that RNA elements that comprise the catalytic core are already formed at the Prp2p stage and that Prp2p destabilizes these elements, directly or indirectly, both to proofread spliceosome activation and to promote reconfiguration of the spliceosome to a fully competent, catalytic conformation. Cold Spring Harbor Laboratory Press 2014-03 /pmc/articles/PMC3923124/ /pubmed/24442613 http://dx.doi.org/10.1261/rna.042598.113 Text en © 2014 Wlodaver and Staley; Published by Cold Spring Harbor Laboratory Press for the RNA Society http://creativecommons.org/licenses/by-nc/3.0/ This article is distributed exclusively by the RNA Society for the first 12 months after the full-issue publication date (see http://rnajournal.cshlp.org/site/misc/terms.xhtml). After 12 months, it is available under a Creative Commons License (Attribution-NonCommercial 3.0 Unported), as described at http://creativecommons.org/licenses/by-nc/3.0/.
spellingShingle Articles
Wlodaver, Alissa M.
Staley, Jonathan P.
The DExD/H-box ATPase Prp2p destabilizes and proofreads the catalytic RNA core of the spliceosome
title The DExD/H-box ATPase Prp2p destabilizes and proofreads the catalytic RNA core of the spliceosome
title_full The DExD/H-box ATPase Prp2p destabilizes and proofreads the catalytic RNA core of the spliceosome
title_fullStr The DExD/H-box ATPase Prp2p destabilizes and proofreads the catalytic RNA core of the spliceosome
title_full_unstemmed The DExD/H-box ATPase Prp2p destabilizes and proofreads the catalytic RNA core of the spliceosome
title_short The DExD/H-box ATPase Prp2p destabilizes and proofreads the catalytic RNA core of the spliceosome
title_sort dexd/h-box atpase prp2p destabilizes and proofreads the catalytic rna core of the spliceosome
topic Articles
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3923124/
https://www.ncbi.nlm.nih.gov/pubmed/24442613
http://dx.doi.org/10.1261/rna.042598.113
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