Cargando…

Evolutionary selection of pestivirus variants with altered or no microRNA dependency

Host microRNA (miRNA) dependency is a hallmark of the human pathogen hepatitis C virus (HCV) and was also described for the related pestiviruses, which are important livestock pathogens. The liver-specific miR-122 binds within the HCV 5′ untranslated region (UTR), whereas the broadly expressed let-7...

Descripción completa

Detalles Bibliográficos
Autores principales: Kokkonos, Konstantinos G, Fossat, Nicolas, Nielsen, Louise, Holm, Christina, Hepkema, Wytske M, Bukh, Jens, Scheel, Troels K H
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Oxford University Press 2020
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7261151/
https://www.ncbi.nlm.nih.gov/pubmed/32374844
http://dx.doi.org/10.1093/nar/gkaa300
_version_ 1783540452251664384
author Kokkonos, Konstantinos G
Fossat, Nicolas
Nielsen, Louise
Holm, Christina
Hepkema, Wytske M
Bukh, Jens
Scheel, Troels K H
author_facet Kokkonos, Konstantinos G
Fossat, Nicolas
Nielsen, Louise
Holm, Christina
Hepkema, Wytske M
Bukh, Jens
Scheel, Troels K H
author_sort Kokkonos, Konstantinos G
collection PubMed
description Host microRNA (miRNA) dependency is a hallmark of the human pathogen hepatitis C virus (HCV) and was also described for the related pestiviruses, which are important livestock pathogens. The liver-specific miR-122 binds within the HCV 5′ untranslated region (UTR), whereas the broadly expressed let-7 and miR-17 families bind two sites (S1 and S2, respectively) in the pestiviral 3′ UTR. Here, we dissected the mechanism of miRNA dependency of the pestivirus bovine viral diarrhea virus (BVDV). Argonaute 2 (AGO2) and miR-17 binding were essential for viral replication, whereas let-7 binding was mainly required for full translational efficiency. Furthermore, using seed site randomized genomes and evolutionary selection experiments, we found that tropism could be redirected to different miRNAs. AGO cross-linking and immunoprecipitation (CLIP) experiments and miRNA antagonism demonstrated that these alternative variants bound and depended on the corresponding miRNAs. Interestingly, we also identified miRNA-independent variants that were obtained through acquisition of compensatory mutations near the genomic 3′ terminus. Rescue experiments demonstrated that miRNA binding and 3′ mutagenesis contribute to replication through mutually exclusive mechanisms. Altogether, our findings suggest that pestiviruses, although capable of miRNA-independent replication, took advantage of miRNAs as essential host factors, suggesting a favorable path during evolutionary adaptation.
format Online
Article
Text
id pubmed-7261151
institution National Center for Biotechnology Information
language English
publishDate 2020
publisher Oxford University Press
record_format MEDLINE/PubMed
spelling pubmed-72611512020-06-03 Evolutionary selection of pestivirus variants with altered or no microRNA dependency Kokkonos, Konstantinos G Fossat, Nicolas Nielsen, Louise Holm, Christina Hepkema, Wytske M Bukh, Jens Scheel, Troels K H Nucleic Acids Res Molecular Biology Host microRNA (miRNA) dependency is a hallmark of the human pathogen hepatitis C virus (HCV) and was also described for the related pestiviruses, which are important livestock pathogens. The liver-specific miR-122 binds within the HCV 5′ untranslated region (UTR), whereas the broadly expressed let-7 and miR-17 families bind two sites (S1 and S2, respectively) in the pestiviral 3′ UTR. Here, we dissected the mechanism of miRNA dependency of the pestivirus bovine viral diarrhea virus (BVDV). Argonaute 2 (AGO2) and miR-17 binding were essential for viral replication, whereas let-7 binding was mainly required for full translational efficiency. Furthermore, using seed site randomized genomes and evolutionary selection experiments, we found that tropism could be redirected to different miRNAs. AGO cross-linking and immunoprecipitation (CLIP) experiments and miRNA antagonism demonstrated that these alternative variants bound and depended on the corresponding miRNAs. Interestingly, we also identified miRNA-independent variants that were obtained through acquisition of compensatory mutations near the genomic 3′ terminus. Rescue experiments demonstrated that miRNA binding and 3′ mutagenesis contribute to replication through mutually exclusive mechanisms. Altogether, our findings suggest that pestiviruses, although capable of miRNA-independent replication, took advantage of miRNAs as essential host factors, suggesting a favorable path during evolutionary adaptation. Oxford University Press 2020-06-04 2020-05-06 /pmc/articles/PMC7261151/ /pubmed/32374844 http://dx.doi.org/10.1093/nar/gkaa300 Text en © The Author(s) 2020. 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 Molecular Biology
Kokkonos, Konstantinos G
Fossat, Nicolas
Nielsen, Louise
Holm, Christina
Hepkema, Wytske M
Bukh, Jens
Scheel, Troels K H
Evolutionary selection of pestivirus variants with altered or no microRNA dependency
title Evolutionary selection of pestivirus variants with altered or no microRNA dependency
title_full Evolutionary selection of pestivirus variants with altered or no microRNA dependency
title_fullStr Evolutionary selection of pestivirus variants with altered or no microRNA dependency
title_full_unstemmed Evolutionary selection of pestivirus variants with altered or no microRNA dependency
title_short Evolutionary selection of pestivirus variants with altered or no microRNA dependency
title_sort evolutionary selection of pestivirus variants with altered or no microrna dependency
topic Molecular Biology
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7261151/
https://www.ncbi.nlm.nih.gov/pubmed/32374844
http://dx.doi.org/10.1093/nar/gkaa300
work_keys_str_mv AT kokkonoskonstantinosg evolutionaryselectionofpestivirusvariantswithalteredornomicrornadependency
AT fossatnicolas evolutionaryselectionofpestivirusvariantswithalteredornomicrornadependency
AT nielsenlouise evolutionaryselectionofpestivirusvariantswithalteredornomicrornadependency
AT holmchristina evolutionaryselectionofpestivirusvariantswithalteredornomicrornadependency
AT hepkemawytskem evolutionaryselectionofpestivirusvariantswithalteredornomicrornadependency
AT bukhjens evolutionaryselectionofpestivirusvariantswithalteredornomicrornadependency
AT scheeltroelskh evolutionaryselectionofpestivirusvariantswithalteredornomicrornadependency