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Inhibition of HCV translation by disrupting the structure and interactions of the viral CRE and 3′ X-tail

A phylogenetically conserved RNA structure within the NS5B coding region of hepatitis C virus functions as a cis-replicating element (CRE). Integrity of this CRE, designated SL9266 (alternatively 5BSL3.2), is critical for genome replication. SL9266 forms the core of an extended pseudoknot, designate...

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Autores principales: Tuplin, Andrew, Struthers, Madeleine, Cook, Jonathan, Bentley, Kirsten, Evans, David J.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Oxford University Press 2015
Materias:
RNA
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4357731/
https://www.ncbi.nlm.nih.gov/pubmed/25712095
http://dx.doi.org/10.1093/nar/gkv142
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author Tuplin, Andrew
Struthers, Madeleine
Cook, Jonathan
Bentley, Kirsten
Evans, David J.
author_facet Tuplin, Andrew
Struthers, Madeleine
Cook, Jonathan
Bentley, Kirsten
Evans, David J.
author_sort Tuplin, Andrew
collection PubMed
description A phylogenetically conserved RNA structure within the NS5B coding region of hepatitis C virus functions as a cis-replicating element (CRE). Integrity of this CRE, designated SL9266 (alternatively 5BSL3.2), is critical for genome replication. SL9266 forms the core of an extended pseudoknot, designated SL9266/PK, involving long distance RNA–RNA interactions between unpaired loops of SL9266 and distal regions of the genome. Previous studies demonstrated that SL9266/PK is dynamic, with ‘open’ and ‘closed’ conformations predicted to have distinct functions during virus replication. Using a combination of site-directed mutagenesis and locked nucleic acids (LNA) complementary to defined domains of SL9266 and its interacting regions, we have explored the influence of this structure on genome translation and replication. We demonstrate that LNAs which block formation of the closed conformation inhibit genome translation. Inhibition was at least partly independent of the initiation mechanism, whether driven by homologous or heterologous internal ribosome entry sites or from a capped message. Provision of SL9266/PK in trans relieved translational inhibition, and mutational analysis implied a mechanism in which the closed conformation recruits a cellular factor that would otherwise suppresses translation. We propose that SL9266/PK functions as a temporal switch, modulating the mutually incompatible processes of translation and replication.
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spelling pubmed-43577312015-03-20 Inhibition of HCV translation by disrupting the structure and interactions of the viral CRE and 3′ X-tail Tuplin, Andrew Struthers, Madeleine Cook, Jonathan Bentley, Kirsten Evans, David J. Nucleic Acids Res RNA A phylogenetically conserved RNA structure within the NS5B coding region of hepatitis C virus functions as a cis-replicating element (CRE). Integrity of this CRE, designated SL9266 (alternatively 5BSL3.2), is critical for genome replication. SL9266 forms the core of an extended pseudoknot, designated SL9266/PK, involving long distance RNA–RNA interactions between unpaired loops of SL9266 and distal regions of the genome. Previous studies demonstrated that SL9266/PK is dynamic, with ‘open’ and ‘closed’ conformations predicted to have distinct functions during virus replication. Using a combination of site-directed mutagenesis and locked nucleic acids (LNA) complementary to defined domains of SL9266 and its interacting regions, we have explored the influence of this structure on genome translation and replication. We demonstrate that LNAs which block formation of the closed conformation inhibit genome translation. Inhibition was at least partly independent of the initiation mechanism, whether driven by homologous or heterologous internal ribosome entry sites or from a capped message. Provision of SL9266/PK in trans relieved translational inhibition, and mutational analysis implied a mechanism in which the closed conformation recruits a cellular factor that would otherwise suppresses translation. We propose that SL9266/PK functions as a temporal switch, modulating the mutually incompatible processes of translation and replication. Oxford University Press 2015-03-11 2015-02-20 /pmc/articles/PMC4357731/ /pubmed/25712095 http://dx.doi.org/10.1093/nar/gkv142 Text en © The Author(s) 2015. Published by Oxford University Press on behalf of Nucleic Acids Research. http://creativecommons.org/licenses/by/4.0/ This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted reuse, distribution, and reproduction in any medium, provided the original work is properly cited.
spellingShingle RNA
Tuplin, Andrew
Struthers, Madeleine
Cook, Jonathan
Bentley, Kirsten
Evans, David J.
Inhibition of HCV translation by disrupting the structure and interactions of the viral CRE and 3′ X-tail
title Inhibition of HCV translation by disrupting the structure and interactions of the viral CRE and 3′ X-tail
title_full Inhibition of HCV translation by disrupting the structure and interactions of the viral CRE and 3′ X-tail
title_fullStr Inhibition of HCV translation by disrupting the structure and interactions of the viral CRE and 3′ X-tail
title_full_unstemmed Inhibition of HCV translation by disrupting the structure and interactions of the viral CRE and 3′ X-tail
title_short Inhibition of HCV translation by disrupting the structure and interactions of the viral CRE and 3′ X-tail
title_sort inhibition of hcv translation by disrupting the structure and interactions of the viral cre and 3′ x-tail
topic RNA
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4357731/
https://www.ncbi.nlm.nih.gov/pubmed/25712095
http://dx.doi.org/10.1093/nar/gkv142
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