Sequence conservation within neutralization epitope regions of VP7 and VP4 proteins of human serotype G4 rotavirus isolates

Abstract

Serotype G4 rotavirus isolates causing four separate epidemics of severe diarrhoea in young children in Melbourne, Australia (from 1974-1990) were investigated for sequence variation in genes encoding the outer capsid proteins, VP4 and VP7. Complementary DNA of the gene encoding the major outer capsid neutralization antigen, VP7, of eighteen isolates was synthesized and amplified by coupled reverse transcription and polymerase chain reaction. Direct sequencing methods were used to derive the deduced amino acid sequences of the immunodominant A, B, and C neutralization epitope regions of the protein. Limited variation was observed among all isolates. A threonine to asparagine change in region A, at amino acid 96, was associated with altered binding of serotype G4-specific neutralizing monoclonal antibodies. The VP8* region of the outer capsid protein VP4 (containing the proposed serotype-specific neutralization epitopes) was investigated in eight isolates. This region was found to highly conserved both within Melbourne isolates and in relation to the standard strains Wa, P, and VA70. The characteristic periodicity of occurrence of serotype G4 isolates causing severe diarrhoea in Melbourne children is unlikely to be due to changes in neutralization epitopes located on the outer capsid proteins, VP7 or VP4.