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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...
Autores principales: | , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Oxford University Press
2015
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Materias: | |
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. |
format | Online Article Text |
id | pubmed-4357731 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2015 |
publisher | Oxford University Press |
record_format | MEDLINE/PubMed |
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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