Logo of aacPermissionsJournals.ASM.orgJournalAAC ArticleJournal InfoAuthorsReviewers
Antimicrob Agents Chemother. 1989 Oct; 33(10): 1668–1673.
PMCID: PMC172735

Molecular analysis of the inhibitory effect of phosphorylated ribavirin on the vesicular stomatitis virus in vitro polymerase reaction.


The effect of phosphorylated ribavirin on the vesicular stomatitis virus in vitro transcription reaction was examined. Analysis of the kinetics observed when the concentrations of nucleoside triphosphates were varied was performed with vesicular stomatitis virus wild-type standard virions. Double-reciprocal and Eadie-Hofstee plots showed competitive inhibition with all natural nucleoside triphosphates when both ribavirin diphosphate (RDP) and ribavirin triphosphate (RTP) were used. The Km values for ATP obtained for the wild-type polymerase were similar to those reported previously. To further characterize the observed inhibition kinetics, in vitro transcription products synthesized in the presence or absence of RDP and RTP were purified by CsCl centrifugation and were primer extended with oligonucleotides specific for either positive-sense leader or nucleocapsid mRNA transcripts. The ratios of leader to nucleocapsid mRNA were measured from primer-extended in vitro transcription products. It was found that the addition of RDP or RTP did not significantly change the in vitro ratio, suggesting that the polymerase is blocked before it enters the 3' end of the template.

Full text

Full text is available as a scanned copy of the original print version. Get a printable copy (PDF file) of the complete article (1.1M), or click on a page image below to browse page by page. Links to PubMed are also available for Selected References.

Images in this article

Click on the image to see a larger version.

Selected References

These references are in PubMed. This may not be the complete list of references from this article.
  • Allen LB, Boswell KH, Khwaja TA, Meyer RB, Jr, Sidwell RW, Witkowski JT, Christensen LF, Robins RK. Synthesis and antiviral acticity of some phosphates of the broad-spectrum antiviral nucleoside, 1-beta-D-ribofuranosyl-1,2,4-triazole-3-carboxamide (ribavirin). J Med Chem. 1978 Aug;21(8):742–746. [PubMed]
  • Bellocq C, Raju R, Patterson J, Kolakofsky D. Translational requirement of La Crosse virus S-mRNA synthesis: in vitro studies. J Virol. 1987 Jan;61(1):87–95. [PMC free article] [PubMed]
  • De BP, Banerjee AK. Requirements and functions of vesicular stomatitis virus L and NS proteins in the transcription process in vitro. Biochem Biophys Res Commun. 1985 Jan 16;126(1):40–49. [PubMed]
  • Eriksson B, Helgstrand E, Johansson NG, Larsson A, Misiorny A, Norén JO, Philipson L, Stenberg K, Stening G, Stridh S, et al. Inhibition of influenza virus ribonucleic acid polymerase by ribavirin triphosphate. Antimicrob Agents Chemother. 1977 Jun;11(6):946–951. [PMC free article] [PubMed]
  • Furman PA, St Clair MH, Spector T. Acyclovir triphosphate is a suicide inactivator of the herpes simplex virus DNA polymerase. J Biol Chem. 1984 Aug 10;259(15):9575–9579. [PubMed]
  • Goswami BB, Borek E, Sharma OK, Fujitaki J, Smith RA. The broad spectrum antiviral agent ribavirin inhibits capping of mRNA. Biochem Biophys Res Commun. 1979 Aug 13;89(3):830–836. [PubMed]
  • Green TL, Emerson SU. Effect of the beta-gamma phosphate bond of ATP on synthesis of leader RNA and mRNAs of vesicular stomatitis virus. J Virol. 1984 Apr;50(1):255–257. [PMC free article] [PubMed]
  • Iverson LE, Rose JK. Localized attenuation and discontinuous synthesis during vesicular stomatitis virus transcription. Cell. 1981 Feb;23(2):477–484. [PubMed]
  • Leppert M, Rittenhouse L, Perrault J, Summers DF, Kolakofsky D. Plus and minus strand leader RNAs in negative strand virus-infected cells. Cell. 1979 Nov;18(3):735–747. [PubMed]
  • Malinoski F, Stollar V. Inhibitors of IMP dehydrogenase prevent sindbis virus replication and reduce GTP levels in Aedes albopictus cells. Virology. 1981 Apr 30;110(2):281–289. [PubMed]
  • Sanger F, Nicklen S, Coulson AR. DNA sequencing with chain-terminating inhibitors. Proc Natl Acad Sci U S A. 1977 Dec;74(12):5463–5467. [PMC free article] [PubMed]
  • Sidwell RW, Huffman JH, Khare GP, Allen LB, Witkowski JT, Robins RK. Broad-spectrum antiviral activity of Virazole: 1-beta-D-ribofuranosyl-1,2,4-triazole-3-carboxamide. Science. 1972 Aug 25;177(4050):705–706. [PubMed]
  • Spector T, Hajian G. Statistical methods to distinguish competitive, noncompetitive, and uncompetitive enzyme inhibitors. Anal Biochem. 1981 Aug;115(2):403–409. [PubMed]
  • Stampfer M, Baltimore D, Huang AS. Ribonucleic acid synthesis of vesicular stomatitis virus. I. Species of ribonucleic acid found in Chinese hamster ovary cells infected with plaque-forming and defective particles. J Virol. 1969 Aug;4(2):154–161. [PMC free article] [PubMed]
  • Streeter DG, Witkowski JT, Khare GP, Sidwell RW, Bauer RJ, Robins RK, Simon LN. Mechanism of action of 1- -D-ribofuranosyl-1,2,4-triazole-3-carboxamide (Virazole), a new broad-spectrum antiviral agent. Proc Natl Acad Sci U S A. 1973 Apr;70(4):1174–1178. [PMC free article] [PubMed]
  • Testa D, Banerjee AK. Initiation of RNA synthesis in vitro by vesicular stomatitis virus. Role of ATP. J Biol Chem. 1979 Mar 25;254(6):2053–2058. [PubMed]
  • Toltzis P, O'Connell K, Patterson JL. Effect of phosphorylated ribavirin on vesicular stomatitis virus transcription. Antimicrob Agents Chemother. 1988 Apr;32(4):492–497. [PMC free article] [PubMed]
  • WILKINSON GN. Statistical estimations in enzyme kinetics. Biochem J. 1961 Aug;80:324–332. [PMC free article] [PubMed]
  • Wray SK, Gilbert BE, Knight V. Effect of ribavirin triphosphate on primer generation and elongation during influenza virus transcription in vitro. Antiviral Res. 1985 Feb;5(1):39–48. [PubMed]
  • Wray SK, Gilbert BE, Noall MW, Knight V. Mode of action of ribavirin: effect of nucleotide pool alterations on influenza virus ribonucleoprotein synthesis. Antiviral Res. 1985 Feb;5(1):29–37. [PubMed]

Articles from Antimicrobial Agents and Chemotherapy are provided here courtesy of American Society for Microbiology (ASM)


Save items

Related citations in PubMed

See reviews...See all...

Cited by other articles in PMC

See all...


  • Compound
    PubChem Compound links
  • MedGen
    Related information in MedGen
  • PubMed
    PubMed citations for these articles
  • Substance
    PubChem Substance links

Recent Activity

Your browsing activity is empty.

Activity recording is turned off.

Turn recording back on

See more...