Murine norovirus (MNV-1) exposure in vitro to the purine nucleoside analog Ribavirin increases quasispecies diversity.
Ribavirin is a pharmaceutical antiviral used for the treatment of RNA virus infections including norovirus, hepatitis C virus, hepatitis E virus, Lassa virus, respiratory syncytial virus, and rhinovirus. Despite the drug's history and documented efficacy, the antiviral mechanism of Ribavirin remains unclear. Mechanisms proposed include depletion of the intracellular GTP pool, immunomodulatory effects, induction of error catastrophe, inhibition of viral polymerase activity, and/or inhibition of viral capping. In the present study, we leveraged deep sequencing data to demonstrate that Ribavirin increases murine norovirus (MNV-1) viral diversity. By serial passaging MNV-1 in RAW 264.7 cells for twenty generations in the presence of Ribavirin, we demonstrated statistically significant increases in both the number of unique haplotypes and the average pairwise difference (APD). Based on statistically significant differences in the probability of nucleotide mutations based on Roche 454 sequencing, we also demonstrated that single nucleotide substitutions are increased in the presence of Ribavirin. Finally, we demonstrated Ribavirin's impact on statistically significantly reducing the relative proportion of the dominant sequence within the quasispecies.
Environmental Health Sciences, Johns Hopkins Bloomberg School of Public Health, Baltimore, MD 21231, United States.
Timothy R
TR
Julian
Department of Molecular Microbiology and Immunology, Johns Hopkins Bloomberg School of Public Health, Baltimore, MD 21231, United States.
Joseph D
JD
Baugher
Department of Microbiology and Immunology, University of Michigan, Ann Arbor, MI 48109, United States.
Christine M
CM
Rippinger
Environmental Health Sciences, Johns Hopkins Bloomberg School of Public Health, Baltimore, MD 21231, United States.
Rebecca
R
Pinekenstein
Department of Microbiology and Immunology, University of Michigan, Ann Arbor, MI 48109, United States.
Abimbola O
AO
Kolawole
Research and Exploratory Development Department, Johns Hopkins University Applied Physics Laboratory, Laurel, MD 20723, United States.
Thomas S
TS
Mehoke
Department of Microbiology and Immunology, University of Michigan, Ann Arbor, MI 48109, United States.
Christiane E
CE
Wobus
Research and Exploratory Development Department, Johns Hopkins University Applied Physics Laboratory, Laurel, MD 20723, United States.
Andrew B
AB
Feldman
Department of Molecular Microbiology and Immunology, Johns Hopkins Bloomberg School of Public Health, Baltimore, MD 21231, United States; Department of Biostatistics, Johns Hopkins Bloomberg School of Public Health, Baltimore, MD 21231, United States.
Fernando J
FJ
Pineda
Environmental Health Sciences, Johns Hopkins Bloomberg School of Public Health, Baltimore, MD 21231, United States. Electronic address: tim.julian@eawag.ch.
Kellogg J
KJ
Schwab
D000818:Animals; D000998:Antiviral Agents / Q000737:chemistry / Q000494:pharmacology*; D002460:Cell Line; D014644:Genetic Variation / Q000187:drug effects; D051379:Mice; D009154:Mutation / Q000187:drug effects; D029322:Norovirus / Q000187:drug effects* / Q000235:genetics*; D011684:Purine Nucleosides / Q000737:chemistry / Q000494:pharmacology*; D012254:Ribavirin / Q000494:pharmacology*
D000998:Antiviral Agents; D011684:Purine Nucleosides; D012254:Ribavirin
MNV-1; Norovirus; Quasispecies; Ribavirin; Serial passaging