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Despite having been used for decades in the treatment of chronic hepatitis C virus (HCV) infection, the precise mechanism of action of RBV is unknown. In other viruses, it inhibits propagation by increasing the rate of G-to-A and C-to-U transitions. Here, we utilized the J6\/JFH1 HCV cell-culture system to investigate whether RBV inhibits HCV through the same mechanism. Infected Huh7.5 cells were treated with increasing concentrations of RBV or its phosphorylated forms. A fragment of the HCV NS5B-polymerase gene was amplified, cloned, and sequenced to estimate genetic distances. We confirm that the antiviral effect of all three RBV-drug forms on HCV relies on induction of specific transitions (G-to-A and C-to-U). These mutations lead to generation of non-infectious virions, reflected by decreased spread of HCV in cell culture despite relatively limited effect on virus genome titers. Moreover, treatment experiments conducted on a novel Huh7.5 cell line stably overexpressing adenosine kinase, a key enzyme for RBV activation, yielded comparable results. This study indicates that RBV action on HCV in hepatoma cell-culture is exerted through increase in mutagenesis, mediated by RBV triphosphate, and leading to production of non-infectious viruses.<\/jats:p>","DOI":"10.1038\/s41598-018-22620-2","type":"journal-article","created":{"date-parts":[[2018,3,9]],"date-time":"2018-03-09T14:39:50Z","timestamp":1520606390000},"update-policy":"https:\/\/doi.org\/10.1007\/springer_crossmark_policy","source":"Crossref","is-referenced-by-count":38,"title":["Antiviral Effect of Ribavirin against HCV Associated with Increased Frequency of G-to-A and C-to-U Transitions in Infectious Cell Culture Model"],"prefix":"10.1038","volume":"8","author":[{"given":"Andrea","family":"Galli","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Helene","family":"Mens","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Judith M.","family":"Gottwein","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jan","family":"Gerstoft","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jens","family":"Bukh","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"297","published-online":{"date-parts":[[2018,3,15]]},"reference":[{"key":"22620_CR1","doi-asserted-by":"publisher","first-page":"703","DOI":"10.1053\/j.gastro.2003.12.002","volume":"126","author":"JM Pawlotsky","year":"2004","unstructured":"Pawlotsky, J. 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Ribavirin: a drug active against many viruses with multiple effects on virus replication and propagation. Molecular basis of ribavirin resistance. Curr Opin Virol 8, 10\u201315, https:\/\/doi.org\/10.1016\/j.coviro.2014.04.011 (2014).","journal-title":"Curr Opin Virol"},{"key":"22620_CR8","doi-asserted-by":"publisher","first-page":"3351","DOI":"10.2174\/092986706778773059","volume":"13","author":"S Russmann","year":"2006","unstructured":"Russmann, S., Grattagliano, I., Portincasa, P., Palmieri, V. O. & Palasciano, G. Ribavirin-induced anemia: mechanisms, risk factors and related targets for future research. Curr Med Chem 13, 3351\u20133357 (2006).","journal-title":"Curr Med Chem"},{"key":"22620_CR9","doi-asserted-by":"publisher","first-page":"1375","DOI":"10.1038\/82191","volume":"6","author":"S Crotty","year":"2000","unstructured":"Crotty, S. et al. The broad-spectrum antiviral ribonucleoside ribavirin is an RNA virus mutagen. 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