Genetic and molecular analyses of spontaneous mutants of human rhinovirus 14 that are resistant to an antiviral compound

B. A. Heinz, R. R. Rueckert, D. A. Shepard, F. J. Dutko, M. A. McKinlay, M. Fancher, M. G. Rossmann, J. Badger, Thomas Smith

Research output: Contribution to journalArticle

87 Citations (Scopus)

Abstract

Spontaneous mutants of human rhinovirus 14 resistant to WIN 52084, an antiviral compound that inhibits attachment to cells, were isolated by selecting plaques that developed when wild-type virus was plated in the presence of high (2 μg/ml) or low (0.1 to 0.4 μg/ml) concentrations of the compound. Two classes of drug resistance were observed: a high-resistance (HR) class with a frequency of about 4 x 10-5, and a low-resistance (LR) class with a 10- to 30-fold-higher frequency. The RNA genomes of 56 HR mutants and 13 LR mutants were sequenced in regions encoding the drug-binding site. The HR mutations mapped to only 2 of the 16 amino acid residues that form the walls of the drug-binding pocket. The side chains of these two residues point directly into the pocket and were invariably replaced by bulkier groups. These findings, and patterns of resistance to related WIN compounds, support the concept that HR mutations may hinder the entry or seating of drug within the binding pocket. In contrast, all of the LR mutations mapped to portions of the polypeptide chain near the canyon floor than move when the drug is inserted. Because several LR mutations partially reverse the attachment-inhibiting effect of WIN compounds, these mutants provide useful tools for studying the regions of the capsid structure involved in attachment. This paper shows that the method of escape mutant analysis, previously used to identify antibody binding sites on human rhinovirus 14, is also applicable to analysis of antiviral drug activity.

Original languageEnglish
Pages (from-to)2476-2485
Number of pages10
JournalJournal of Virology
Volume63
Issue number6
StatePublished - 1989
Externally publishedYes

Fingerprint

Human rhinovirus
Rhinovirus
Antiviral Agents
Molecular Biology
mutants
Mutation
mutation
drugs
Pharmaceutical Preparations
Antibody Binding Sites
Capsid
antiviral agents
Drug Resistance
capsid
canyons
drug resistance
Binding Sites
Genome
RNA
binding sites

ASJC Scopus subject areas

  • Immunology

Cite this

Heinz, B. A., Rueckert, R. R., Shepard, D. A., Dutko, F. J., McKinlay, M. A., Fancher, M., ... Smith, T. (1989). Genetic and molecular analyses of spontaneous mutants of human rhinovirus 14 that are resistant to an antiviral compound. Journal of Virology, 63(6), 2476-2485.

Genetic and molecular analyses of spontaneous mutants of human rhinovirus 14 that are resistant to an antiviral compound. / Heinz, B. A.; Rueckert, R. R.; Shepard, D. A.; Dutko, F. J.; McKinlay, M. A.; Fancher, M.; Rossmann, M. G.; Badger, J.; Smith, Thomas.

In: Journal of Virology, Vol. 63, No. 6, 1989, p. 2476-2485.

Research output: Contribution to journalArticle

Heinz, BA, Rueckert, RR, Shepard, DA, Dutko, FJ, McKinlay, MA, Fancher, M, Rossmann, MG, Badger, J & Smith, T 1989, 'Genetic and molecular analyses of spontaneous mutants of human rhinovirus 14 that are resistant to an antiviral compound', Journal of Virology, vol. 63, no. 6, pp. 2476-2485.
Heinz BA, Rueckert RR, Shepard DA, Dutko FJ, McKinlay MA, Fancher M et al. Genetic and molecular analyses of spontaneous mutants of human rhinovirus 14 that are resistant to an antiviral compound. Journal of Virology. 1989;63(6):2476-2485.
Heinz, B. A. ; Rueckert, R. R. ; Shepard, D. A. ; Dutko, F. J. ; McKinlay, M. A. ; Fancher, M. ; Rossmann, M. G. ; Badger, J. ; Smith, Thomas. / Genetic and molecular analyses of spontaneous mutants of human rhinovirus 14 that are resistant to an antiviral compound. In: Journal of Virology. 1989 ; Vol. 63, No. 6. pp. 2476-2485.
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