Elsevier

Virus Research

Volume 107, Issue 2, February 2005, Pages 173-181
Virus Research

Ribavirin and lethal mutagenesis of poliovirus: molecular mechanisms, resistance and biological implications

https://doi.org/10.1016/j.virusres.2004.11.007Get rights and content

Abstract

Positive strand RNA virus populations are a collection of similar but genetically different viruses. They exist as viral quasispecies due to the high mutation rates of the low fidelity viral RNA-dependent RNA polymerase (RdRp). It is thought that this genomic heterogeneity is advantageous to the population, allowing for adaptation to rapidly changing environments that present varying types and degrees of selective pressure. However, one consequence of this extensive diversity is the susceptibility to mutagens that further increase sequence variation. Since RNA viruses live at the edge of maximal variability, an increase in the mutation rate is likely to force the virus beyond the tolerable mutation frequency into ‘error catastrophe’. One such mutagen, ribavirin, is an antiviral nucleoside analog that is mutagenic to several RNA viruses. Ribavirin is incorporated into the viral genome causing lethal mutagenesis and a subsequent decrease in the specific infectivity. Even so, passaging poliovirus in the presence of low to intermediate concentrations of the drug leads to the emergence of a viral population resistant to the effects of ribavirin. These viruses have a point mutation in the RdRp that increases the overall polymerase fidelity. Interestingly, as predicted by the quasispecies theory, ribavirin resistant viruses are less adaptable, as they are more susceptible to other non-mutagenic antiviral drugs and are highly attenuated in vivo. Here, we review the mechanism of action of ribavirin on poliovirus and other RNA viruses, the possibility for escape via increased fidelity of the viral polymerase, the consequences of this response on viral population dynamics, and the biological implications for the therapeutic use of mutagenic antiviral agents.

Section snippets

Poliovirus exists on the edge of error catastrophe: the safety of living dangerously

Poliovirus, the prototype picornavirus, is a non-enveloped virus with a single stranded RNA genome of positive polarity. The virion is composed of an icosahedral protein shell composed of four capsid proteins VP1, VP2, VP3 and VP4 encompassing the RNA genome. The genome of approximately 7.5 kb contains a single ORF encoding a polyprotein that is cleaved into the individual viral proteins required for virus replication and assembly. Poliovirus has a characteristically rapid multiplication cycle

Ribavirin induces lethal mutagenesis of poliovirus

Ribavirin has a broad-spectrum antiviral activity against both RNA and DNA viruses. A number of viruses in different taxonomic viral families are susceptible to ribavirin (summarized in Table 1). Yet, the mechanisms by which ribavirin inhibits these different viruses has been difficult to elucidate, as this drug has pleiotropic effects. The inhibition of cellular inosine monophosphate dehydrogenase (IMPDH) and subsequent decrease of intracellular GTP levels can reduce viral protein translation

A high fidelity polymerase confers resistance to lethal mutagenesis

Indeed, passage of poliovirus in ribavirin treated cells resulted in the acquisition of a RdRp mutant with a glycine to serine mutation at position 64 (G64S) of the polymerase (Pfeiffer and Kirkegaard, 2003, Vignuzzi et al., 2004). Interestingly, examination of the known crystal-derived structure (Hansen et al., 1997) of the poliovirus RdRp revealed that this mutation was not found within any of the known functional domains but was situated within the small region spanning the finger and thumb

Ribavirin and RNA mutagens as antivirals: biological implications for diverse virus families

As highlighted above, ribavirin is a potent antiviral that exerts its effects through lethal mutagenesis. The poliovirus model is an excellent system for these studies, as the effects of RNA mutagens can be readily measured and compared in tissue culture or in animal models. Furthermore, established biochemical approaches permit analysis of parameters useful in determining ideal RNA mutagens. Ribavirin analogs can be developed and quickly tested in such a system, in order to identify which

Acknowledgements

This work was financially supported by an NIH fellowship to J.S. and by NIH grant AI40085 to R.A. We would like to thank R. Van Rij, C. Ruiz-Jarabo, and O. Peersen for helpful discussions.

References (93)

  • S.H. Fang et al.

    Ribavirin enhancement of hepatitis C virus core antigen-specific type 1 T helper cell response correlates with the increased IL-12 level

    J. Hepatol.

    (2000)
  • D.W. Gohara et al.

    Poliovirus RNA-dependent RNA polymerase (3Dpol): structural, biochemical, and biological analysis of conserved structural motifs A and B

    J. Biol. Chem.

    (2000)
  • J.L. Hansen et al.

    Structure of the RNA-dependent RNA polymerase of poliovirus

    Structure

    (1997)
  • J.H. Lee et al.

    Effect of ribavirin on virus load and quasispecies distribution in patients infected with hepatitis C virus

    J. Hepatol.

    (1998)
  • D. Maag et al.

    Hepatitis C virus RNA-dependent RNA polymerase (NS5B) as a mediator of the antiviral activity of ribavirin

    J. Biol. Chem.

    (2001)
  • J.B. McCormick et al.

    Ribavirin suppresses replication of lymphadenopathy-associated virus in cultures of human adult T lymphocytes

    Lancet

    (1984)
  • A. Meyerhans et al.

    The fidelity of cellular and viral polymerases and its manipulation for hypermutagenesis

  • R.K. Robins et al.

    The importance of IMP dehydrogenase inhibition in the broad spectrum antiviral activity of ribavirin and selenazofurin

    Adv. Enzyme Regul.

    (1985)
  • C.M. Ruiz-Jarabo et al.

    Lethal mutagenesis of the prototypic arenavirus lymphocytic choriomeningitis virus (LCMV)

    Virology

    (2003)
  • D.F. Smee et al.

    Selective inhibition of arthropod-borne and arenaviruses in vitro by 3’-fluoro-3’-deoxyadenosine

    Antiviral Res.

    (1992)
  • D.A. Steinhauer et al.

    Lack of evidence for proofreading mechanisms associated with an RNA virus polymerase

    Gene

    (1992)
  • E.L. Stephen et al.

    Experimental Lassa fever virus infection successfully treated with ribavirin

    Lancet

    (1979)
  • T. Takahashi et al.

    The cooperative effect of interferon-alpha and ribavirin on subacute sclerosing panencephalitis (SSPE) virus infections, in vitro and in vivo

    Antiviral Res.

    (1998)
  • R.C. Tam et al.

    Ribavirin polarizes human T cell responses towards a Type 1 cytokine profile

    J. Hepatol.

    (1999)
  • F.H. van Tiel et al.

    Inhibition of Semliki Forest virus multiplication by ribavirin: a potential method for the monitoring of antiviral agents in serum

    J. Virol. Methods

    (1986)
  • S.K. Wray et al.

    Effect of ribavirin triphosphate on primer generation and elongation during influenza virus transcription in vitro

    Antiviral Res.

    (1985)
  • S.K. Wray et al.

    Mode of action of ribavirin: effect of nucleotide pool alterations on influenza virus ribonucleoprotein synthesis

    Antiviral Res.

    (1985)
  • P.R. Wyde

    Respiratory syncytial virus (RSV) disease and prospects for its control

    Antiviral Res.

    (1998)
  • P.R. Wyde et al.

    Comparison of the inhibition of human metapneumovirus and respiratory syncytial virus by ribavirin and immune serum globulin in vitro

    Antiviral Res.

    (2003)
  • K.C. Young et al.

    Identification of a ribavirin-resistant NS5B mutation of hepatitis C virus during ribavirin monotherapy

    Hepatology

    (2003)
  • S. Zhou et al.

    The effect of ribavirin and IMPDH inhibitors on hepatitis C virus subgenomic replicon RNA

    Virology

    (2003)
  • C.E. Cameron et al.

    The mechanism of action of ribavirin: lethal mutagenesis of RNA virus genomes mediated by the viral RNA-dependent RNA polymerase

    Curr. Opin. Infect. Dis.

    (2001)
  • A.M. Contreras et al.

    Viral RNA mutations are region specific and increased by ribavirin in a full-length hepatitis C virus replication system

    J. Virol.

    (2002)
  • S. Crotty et al.

    Ribavirin's antiviral mechanism of action: lethal mutagenesis?

    J. Mol. Med.

    (2002)
  • S. Crotty et al.

    RNA virus error catastrophe: direct molecular test by using ribavirin.

    Proc. Natl. Acad. Sci. U.S.A.

    (2001)
  • S. Crotty et al.

    The broad-spectrum antiviral ribonucleoside ribavirin is an RNA virus mutagen

    Nat. Med.

    (2000)
  • E. Domingo

    Rapid evolution of viral RNA genomes

    J. Nutr.

    (2000)
  • E. Domingo et al.

    Quasispecies and molecular evolution of viruses

    Rev. Sci. Tech.

    (2000)
  • E. Domingo et al.

    Quasispecies structure and persistence of RNA viruses

    Emerg. Infect. Dis.

    (1998)
  • E. Domingo et al.

    Basic concepts in RNA virus evolution

    FASEB J.

    (1996)
  • E. Domingo et al.

    RNA virus mutations and fitness for survival

    Annu. Rev. Microbiol.

    (1997)
  • E. Domingo et al.

    Mutation rates and rapid evolution of RNA viruses

  • E. Domingo et al.

    RNA Genetics

    (1988)
  • E. Domingo et al.

    RNA virus fitness

    Rev. Med. Virol.

    (1997)
  • E. Domingo et al.

    Viral quasispecies and the problem of vaccine-escape and drug-resistant mutants

    Prog. Drug Res.

    (1997)
  • J.W. Drake

    A constant rate of spontaneous mutation in DNA-based microbes

    Proc. Natl. Acad. Sci. U.S.A.

    (1991)
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    Both authors contributed equally to this review.

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