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A homolog of lariat-debranching enzyme modulates turnover of branched RNA
Turnover of the branched RNA intermediates and products of pre-mRNA splicing is mediated by the lariat-debranching enzyme Dbr1. We characterized a homolog of Dbr1 from Saccharomyces cerevisiae, Drn1/Ygr093w, that has a pseudo-metallophosphodiesterase domain with primary sequence homology to Dbr1 but...
Autores principales: | , , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Cold Spring Harbor Laboratory Press
2014
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4105757/ https://www.ncbi.nlm.nih.gov/pubmed/24919400 http://dx.doi.org/10.1261/rna.044602.114 |
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author | Garrey, Stephen M. Katolik, Adam Prekeris, Mantas Li, Xueni York, Kerri Bernards, Sarah Fields, Stanley Zhao, Rui Damha, Masad J. Hesselberth, Jay R. |
author_facet | Garrey, Stephen M. Katolik, Adam Prekeris, Mantas Li, Xueni York, Kerri Bernards, Sarah Fields, Stanley Zhao, Rui Damha, Masad J. Hesselberth, Jay R. |
author_sort | Garrey, Stephen M. |
collection | PubMed |
description | Turnover of the branched RNA intermediates and products of pre-mRNA splicing is mediated by the lariat-debranching enzyme Dbr1. We characterized a homolog of Dbr1 from Saccharomyces cerevisiae, Drn1/Ygr093w, that has a pseudo-metallophosphodiesterase domain with primary sequence homology to Dbr1 but lacks essential active site residues found in Dbr1. Whereas loss of Dbr1 results in lariat-introns failing broadly to turnover, loss of Drn1 causes low levels of lariat-intron accumulation. Conserved residues in the Drn1 C-terminal CwfJ domains, which are not present in Dbr1, are required for efficient intron turnover. Drn1 interacts with Dbr1, components of the Nineteen Complex, U2 snRNA, branched intermediates, and products of splicing. Drn1 enhances debranching catalyzed by Dbr1 in vitro, but does so without significantly improving the affinity of Dbr1 for branched RNA. Splicing carried out in in vitro extracts in the absence of Drn1 results in an accumulation of branched splicing intermediates and products released from the spliceosome, likely due to less active debranching, as well as the promiscuous release of cleaved 5′-exon. Drn1 enhances Dbr1-mediated turnover of lariat-intermediates and lariat-intron products, indicating that branched RNA turnover is regulated at multiple steps during splicing. |
format | Online Article Text |
id | pubmed-4105757 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2014 |
publisher | Cold Spring Harbor Laboratory Press |
record_format | MEDLINE/PubMed |
spelling | pubmed-41057572015-08-01 A homolog of lariat-debranching enzyme modulates turnover of branched RNA Garrey, Stephen M. Katolik, Adam Prekeris, Mantas Li, Xueni York, Kerri Bernards, Sarah Fields, Stanley Zhao, Rui Damha, Masad J. Hesselberth, Jay R. RNA Articles Turnover of the branched RNA intermediates and products of pre-mRNA splicing is mediated by the lariat-debranching enzyme Dbr1. We characterized a homolog of Dbr1 from Saccharomyces cerevisiae, Drn1/Ygr093w, that has a pseudo-metallophosphodiesterase domain with primary sequence homology to Dbr1 but lacks essential active site residues found in Dbr1. Whereas loss of Dbr1 results in lariat-introns failing broadly to turnover, loss of Drn1 causes low levels of lariat-intron accumulation. Conserved residues in the Drn1 C-terminal CwfJ domains, which are not present in Dbr1, are required for efficient intron turnover. Drn1 interacts with Dbr1, components of the Nineteen Complex, U2 snRNA, branched intermediates, and products of splicing. Drn1 enhances debranching catalyzed by Dbr1 in vitro, but does so without significantly improving the affinity of Dbr1 for branched RNA. Splicing carried out in in vitro extracts in the absence of Drn1 results in an accumulation of branched splicing intermediates and products released from the spliceosome, likely due to less active debranching, as well as the promiscuous release of cleaved 5′-exon. Drn1 enhances Dbr1-mediated turnover of lariat-intermediates and lariat-intron products, indicating that branched RNA turnover is regulated at multiple steps during splicing. Cold Spring Harbor Laboratory Press 2014-08 /pmc/articles/PMC4105757/ /pubmed/24919400 http://dx.doi.org/10.1261/rna.044602.114 Text en © 2014 Garrey 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 | Articles Garrey, Stephen M. Katolik, Adam Prekeris, Mantas Li, Xueni York, Kerri Bernards, Sarah Fields, Stanley Zhao, Rui Damha, Masad J. Hesselberth, Jay R. A homolog of lariat-debranching enzyme modulates turnover of branched RNA |
title | A homolog of lariat-debranching enzyme modulates turnover of branched RNA |
title_full | A homolog of lariat-debranching enzyme modulates turnover of branched RNA |
title_fullStr | A homolog of lariat-debranching enzyme modulates turnover of branched RNA |
title_full_unstemmed | A homolog of lariat-debranching enzyme modulates turnover of branched RNA |
title_short | A homolog of lariat-debranching enzyme modulates turnover of branched RNA |
title_sort | homolog of lariat-debranching enzyme modulates turnover of branched rna |
topic | Articles |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4105757/ https://www.ncbi.nlm.nih.gov/pubmed/24919400 http://dx.doi.org/10.1261/rna.044602.114 |
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