title: Isolation and characterization of NMSO3-resistant mutants of respiratory syncytial virus.
abstract: We obtained two mutant strains of respiratory syncytial virus (RSV) which showed resistance against NMSO3 after 15 and 33 passages, respectively, in HEp-2 cells in the presence of 6.8 microM of NMSO3. The EC50 values of NMSO3 for the resistant virus strains were 0.48 and 0.93 microM, that is 4.8-9.3 times higher than that of the parent strain (EC50 = 0.1 microM). The most resistant strain also showed resistance against heparin but was sensitive to dextran sulfate and a polyoxotungstate, PM-523. In order to determine whether the acquisition of resistance to NMSO3 was the result of the accumulation of genetic changes of virus, we sequenced the G- and F-protein genes. In comparison with the standard type of RSV strains, we identified changes of 10 amino acids in the G protein including those at the central conserved segment. However, we did not observe any particular changes in the amino acid sequence of the F-protein of the resistant strains. From these results, we conclude that NMSO3 inhibits the G-protein interaction to the receptor. The mutations in the G-protein may result in the observed phenotypic resistance of RSV towards NMSO3.
authors.affiliation: Department of Microbiology, Fukushima Medical University School of Medicine, 1, Hikarigaoka, Fukushima 960-1295, Japan.
authors.forename: Kazufumi
authors.initials: K
authors.lastname: Kimura
authors.forename: Ken
authors.initials: K
authors.lastname: Ishioka
authors.forename: Koichi
authors.initials: K
authors.lastname: Hashimoto
authors.forename: Schuichi
authors.initials: S
authors.lastname: Mori
authors.forename: Tatsuo
authors.initials: T
authors.lastname: Suzutani
authors.forename: Terry L
authors.initials: TL
authors.lastname: Bowlin
authors.forename: Shiro
authors.initials: S
authors.lastname: Shigeta
mesh_terms: D000998:Antiviral Agents / Q000494:pharmacology*; D045744:Cell Line, Tumor; D017124:Conserved Sequence; D004252:DNA Mutational Analysis; D016264:Dextran Sulfate / Q000494:pharmacology; D024882:Drug Resistance, Viral / Q000235:genetics*; D006493:Heparin / Q000494:pharmacology; D006801:Humans; D008055:Lipids / Q000494:pharmacology*; D020125:Mutation, Missense; D019158:N-Acetylneuraminic Acid / Q000031:analogs & derivatives* / Q000494:pharmacology*; D021141:Nucleic Acid Amplification Techniques; D012367:RNA, Viral / Q000302:isolation & purification; D012136:Respiratory Syncytial Viruses / Q000187:drug effects* / Q000235:genetics* / Q000254:growth & development / Q000302:isolation & purification; D014764:Viral Proteins / Q000235:genetics
chemical_list: C413863:(2,2-bis(docosyloxymethyl)propyl-5-acetamido-3,5-dideoxy-4,7,8,9-tetra-O-(sodium oxysulfonyl)glycero-alpha-galacto-2-nonulopyranosid)onate; D000998:Antiviral Agents; D008055:Lipids; D012367:RNA, Viral; D014764:Viral Proteins; D006493:Heparin; D016264:Dextran Sulfate; D019158:N-Acetylneuraminic Acid