Evolutionary selection of pestivirus variants with altered or no microRNA dependency
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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.
In: Nucleic Acids Research, Vol. 48, No. 10, 04.06.2020, p. 5555-5571.Research output: Contribution to journal › Journal article › Research › peer-review
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TY - JOUR
T1 - Evolutionary selection of pestivirus variants with altered or no microRNA dependency
AU - Kokkonos, Konstantinos G.
AU - Fossat, Nicolas
AU - Nielsen, Louise
AU - Holm, Christina
AU - Hepkema, Wytske M.
AU - Bukh, Jens
AU - Scheel, Troels K.H.
PY - 2020/6/4
Y1 - 2020/6/4
N2 - 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.
AB - 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.
U2 - 10.1093/nar/gkaa300
DO - 10.1093/nar/gkaa300
M3 - Journal article
C2 - 32374844
AN - SCOPUS:85085905159
VL - 48
SP - 5555
EP - 5571
JO - Nucleic Acids Research
JF - Nucleic Acids Research
SN - 0305-1048
IS - 10
ER -
ID: 242795149