title: A single amino acid substitution in the capsid protein VP1 of coxsackievirus B3 (CVB3) alters plaque phenotype in Vero cells but not cardiovirulence in a mouse model.
abstract: We previously described a large plaque attenuant (p14V-1) derived from a cardiovirulent Coxsackievirus B3 (CVB3) and showed that there were no major determinants of either attenuation or plaque phenotype in the 5' nontranslated region (5'NTR). Part of the region encoding the last 124 amino acids of VP3 and the first 106 amino acids of VP1 of the attenuant was then sequenced and compared to the wild-type. Three nucleotide changes were found in the VP1 coding region: a silent single base change at nucleotide position 2467 (C to U) and a double-base change at position 2690-1 (AA to GT), which leads to a change from lysine to serine at amino acid position 80. This mutation maps to the begining of B-C loop of the three-dimensional structure of VP1 of CVB3, where a distinct surface projection is formed. Two infectious chimeric cDNA clones were constructed, based on a cardiovirulent cDNA construct. In one construct, the 5'NTR and the VP3-VP1 region were from p14V-1 and in the other, only the VP3-VP1 region was from this attenuant. Both chimeric viruses produced large plaques on Vero cell monolayers, similar to p14V-1 but larger than the prototypic cardiovirulent virus. In vivo experiments showed that both chimeric viruses induced myocarditis in a murine model, similar to wild-type virus. We conclude that mutation serine-80 in capsid protein VP1 of p14V-1 is a determinant of the large plaque phenotype but is not responsible for attenuation.
authors.affiliation: Department of Biochemistry, Charing Cross and Westminster Medical School, London, U.K.
authors.forename: H
authors.initials: H
authors.lastname: Zhang
authors.forename: N W
authors.initials: NW
authors.lastname: Blake
authors.forename: X
authors.initials: X
authors.lastname: Ouyang
authors.forename: Y A
authors.initials: YA
authors.lastname: Pandolfino
authors.forename: P
authors.initials: P
authors.lastname: Morgan-Capner
authors.forename: L C
authors.initials: LC
authors.lastname: Archard
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D002213:Capsid / Q000737:chemistry / Q000502:physiology*; D036022:Capsid Proteins; D002522:Chlorocebus aethiops; D029822:Enterovirus B, Human / Q000254:growth & development* / Q000472:pathogenicity; D051379:Mice; D008969:Molecular Sequence Data; D009205:Myocarditis / Q000209:etiology*; D013329:Structure-Activity Relationship; D014709:Vero Cells; D010948:Viral Plaque Assay; D014774:Virulence
chemical_list: D036022:Capsid Proteins