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Yeast Prp2 liberates the 5′ splice site and the branch site adenosine for catalysis of pre-mRNA splicing
The RNA helicase Prp2 facilitates the remodeling of the spliceosomal B(act) complex to the catalytically activated B* complex just before step one of splicing. As a high-resolution cryo-EM structure of the B* complex is currently lacking, the precise spliceosome remodeling events mediated by Prp2 re...
Autores principales: | , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Cold Spring Harbor Laboratory Press
2017
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5688998/ https://www.ncbi.nlm.nih.gov/pubmed/28864812 http://dx.doi.org/10.1261/rna.063115.117 |
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author | Bao, Penghui Höbartner, Claudia Hartmuth, Klaus Lührmann, Reinhard |
author_facet | Bao, Penghui Höbartner, Claudia Hartmuth, Klaus Lührmann, Reinhard |
author_sort | Bao, Penghui |
collection | PubMed |
description | The RNA helicase Prp2 facilitates the remodeling of the spliceosomal B(act) complex to the catalytically activated B* complex just before step one of splicing. As a high-resolution cryo-EM structure of the B* complex is currently lacking, the precise spliceosome remodeling events mediated by Prp2 remain poorly understood. To investigate the latter, we used chemical structure probing to compare the RNA structure of purified yeast B(act) and B* complexes. Our studies reveal deviations from conventional RNA helices in the functionally important U6 snRNA internal stem–loop and U2/U6 helix Ib in the activated B(act) complex, and to a lesser extent in B*. Interestingly, the N7 of U6–G60 of the catalytic triad becomes accessible to DMS modification in the B* complex, suggesting that the Hoogsteen interaction with U6–A52 is destabilized in B*. Our data show that Prp2 action does not unwind double-stranded RNA, but enhances the flexibility of the first step reactants, the pre-mRNA's 5′ splice site and branch site adenosine. Prp2 therefore appears to act primarily as an RNPase to achieve catalytic activation by liberating the first step reactants in preparation for catalysis of the first step of splicing. |
format | Online Article Text |
id | pubmed-5688998 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2017 |
publisher | Cold Spring Harbor Laboratory Press |
record_format | MEDLINE/PubMed |
spelling | pubmed-56889982018-12-01 Yeast Prp2 liberates the 5′ splice site and the branch site adenosine for catalysis of pre-mRNA splicing Bao, Penghui Höbartner, Claudia Hartmuth, Klaus Lührmann, Reinhard RNA Report The RNA helicase Prp2 facilitates the remodeling of the spliceosomal B(act) complex to the catalytically activated B* complex just before step one of splicing. As a high-resolution cryo-EM structure of the B* complex is currently lacking, the precise spliceosome remodeling events mediated by Prp2 remain poorly understood. To investigate the latter, we used chemical structure probing to compare the RNA structure of purified yeast B(act) and B* complexes. Our studies reveal deviations from conventional RNA helices in the functionally important U6 snRNA internal stem–loop and U2/U6 helix Ib in the activated B(act) complex, and to a lesser extent in B*. Interestingly, the N7 of U6–G60 of the catalytic triad becomes accessible to DMS modification in the B* complex, suggesting that the Hoogsteen interaction with U6–A52 is destabilized in B*. Our data show that Prp2 action does not unwind double-stranded RNA, but enhances the flexibility of the first step reactants, the pre-mRNA's 5′ splice site and branch site adenosine. Prp2 therefore appears to act primarily as an RNPase to achieve catalytic activation by liberating the first step reactants in preparation for catalysis of the first step of splicing. Cold Spring Harbor Laboratory Press 2017-12 /pmc/articles/PMC5688998/ /pubmed/28864812 http://dx.doi.org/10.1261/rna.063115.117 Text en © 2017 Bao et al.; Published by Cold Spring Harbor Laboratory Press for the RNA Society http://creativecommons.org/licenses/by-nc/4.0/ 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 4.0 International), as described at http://creativecommons.org/licenses/by-nc/4.0/. |
spellingShingle | Report Bao, Penghui Höbartner, Claudia Hartmuth, Klaus Lührmann, Reinhard Yeast Prp2 liberates the 5′ splice site and the branch site adenosine for catalysis of pre-mRNA splicing |
title | Yeast Prp2 liberates the 5′ splice site and the branch site adenosine for catalysis of pre-mRNA splicing |
title_full | Yeast Prp2 liberates the 5′ splice site and the branch site adenosine for catalysis of pre-mRNA splicing |
title_fullStr | Yeast Prp2 liberates the 5′ splice site and the branch site adenosine for catalysis of pre-mRNA splicing |
title_full_unstemmed | Yeast Prp2 liberates the 5′ splice site and the branch site adenosine for catalysis of pre-mRNA splicing |
title_short | Yeast Prp2 liberates the 5′ splice site and the branch site adenosine for catalysis of pre-mRNA splicing |
title_sort | yeast prp2 liberates the 5′ splice site and the branch site adenosine for catalysis of pre-mrna splicing |
topic | Report |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5688998/ https://www.ncbi.nlm.nih.gov/pubmed/28864812 http://dx.doi.org/10.1261/rna.063115.117 |
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