Resurrection of a Viral Internal Ribosome Entry Site from a 700 Year Old Ancient Northwest Territories Cripavirus
The dicistrovirus intergenic region internal ribosome entry site (IGR IRES) uses an unprecedented, streamlined mechanism whereby the IRES adopts a triple-pseudoknot (PK) structure to directly bind to the conserved core of the ribosome and drive translation from a non-AUG codon. The origin of this IR...
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ftpubmed:oai:pubmedcentral.nih.gov:8002689 2023-05-15T17:46:33+02:00 Resurrection of a Viral Internal Ribosome Entry Site from a 700 Year Old Ancient Northwest Territories Cripavirus Wang, Xinying Vlok, Marli Flibotte, Stephane Jan, Eric 2021-03-17 http://www.ncbi.nlm.nih.gov/pmc/articles/PMC8002689/ https://doi.org/10.3390/v13030493 en eng MDPI http://www.ncbi.nlm.nih.gov/pmc/articles/PMC8002689/ http://dx.doi.org/10.3390/v13030493 © 2021 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/). CC-BY Viruses Article Text 2021 ftpubmed https://doi.org/10.3390/v13030493 2021-04-04T01:07:32Z The dicistrovirus intergenic region internal ribosome entry site (IGR IRES) uses an unprecedented, streamlined mechanism whereby the IRES adopts a triple-pseudoknot (PK) structure to directly bind to the conserved core of the ribosome and drive translation from a non-AUG codon. The origin of this IRES mechanism is not known. Previously, a partial fragment of a divergent dicistrovirus RNA genome, named ancient Northwest territories cripavirus (aNCV), was extracted from 700-year-old caribou feces trapped in a subarctic ice patch. The aNCV IGR sequence adopts a secondary structure similar to contemporary IGR IRES structures, however, there are subtle differences including 105 nucleotides upstream of the IRES of unknown function. Using filter binding assays, we showed that the aNCV IRES could bind to purified ribosomes, and toeprinting analysis pinpointed the start site at a GCU alanine codon adjacent to PKI. Using a bicistronic reporter RNA, the aNCV IGR can direct translation in vitro in a PKI-dependent manner. Lastly, a chimeric infectious clone swapping in the aNCV IRES supported translation and virus infection. The characterization and resurrection of a functional IGR IRES from a divergent 700-year-old virus provides a historical framework for the importance of this viral translational mechanism. Text Northwest Territories Subarctic PubMed Central (PMC) Northwest Territories Viruses 13 3 493 |
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Article Wang, Xinying Vlok, Marli Flibotte, Stephane Jan, Eric Resurrection of a Viral Internal Ribosome Entry Site from a 700 Year Old Ancient Northwest Territories Cripavirus |
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description |
The dicistrovirus intergenic region internal ribosome entry site (IGR IRES) uses an unprecedented, streamlined mechanism whereby the IRES adopts a triple-pseudoknot (PK) structure to directly bind to the conserved core of the ribosome and drive translation from a non-AUG codon. The origin of this IRES mechanism is not known. Previously, a partial fragment of a divergent dicistrovirus RNA genome, named ancient Northwest territories cripavirus (aNCV), was extracted from 700-year-old caribou feces trapped in a subarctic ice patch. The aNCV IGR sequence adopts a secondary structure similar to contemporary IGR IRES structures, however, there are subtle differences including 105 nucleotides upstream of the IRES of unknown function. Using filter binding assays, we showed that the aNCV IRES could bind to purified ribosomes, and toeprinting analysis pinpointed the start site at a GCU alanine codon adjacent to PKI. Using a bicistronic reporter RNA, the aNCV IGR can direct translation in vitro in a PKI-dependent manner. Lastly, a chimeric infectious clone swapping in the aNCV IRES supported translation and virus infection. The characterization and resurrection of a functional IGR IRES from a divergent 700-year-old virus provides a historical framework for the importance of this viral translational mechanism. |
format |
Text |
author |
Wang, Xinying Vlok, Marli Flibotte, Stephane Jan, Eric |
author_facet |
Wang, Xinying Vlok, Marli Flibotte, Stephane Jan, Eric |
author_sort |
Wang, Xinying |
title |
Resurrection of a Viral Internal Ribosome Entry Site from a 700 Year Old Ancient Northwest Territories Cripavirus |
title_short |
Resurrection of a Viral Internal Ribosome Entry Site from a 700 Year Old Ancient Northwest Territories Cripavirus |
title_full |
Resurrection of a Viral Internal Ribosome Entry Site from a 700 Year Old Ancient Northwest Territories Cripavirus |
title_fullStr |
Resurrection of a Viral Internal Ribosome Entry Site from a 700 Year Old Ancient Northwest Territories Cripavirus |
title_full_unstemmed |
Resurrection of a Viral Internal Ribosome Entry Site from a 700 Year Old Ancient Northwest Territories Cripavirus |
title_sort |
resurrection of a viral internal ribosome entry site from a 700 year old ancient northwest territories cripavirus |
publisher |
MDPI |
publishDate |
2021 |
url |
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC8002689/ https://doi.org/10.3390/v13030493 |
geographic |
Northwest Territories |
geographic_facet |
Northwest Territories |
genre |
Northwest Territories Subarctic |
genre_facet |
Northwest Territories Subarctic |
op_source |
Viruses |
op_relation |
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC8002689/ http://dx.doi.org/10.3390/v13030493 |
op_rights |
© 2021 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/). |
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CC-BY |
op_doi |
https://doi.org/10.3390/v13030493 |
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Viruses |
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13 |
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493 |
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