Molecular Characterization of the Outer Capsid Spike Protein (VP4) Gene from Human Group C Rotavirus

Abstract

A cDNA copy of the third genomic RNA segment of a noncultivatable human group C rotavirus (Bristol) was generated by single primer amplification. Human group C rotavirus genome segment 3 contains 2283 bp and encodes the VP4 gene with an open reading frame of 2232 nucleotides (744 amino acids) starting at nucleotide 21 and terminating at nucleotide 2251. PCR primers designed from the 5′ and 3′ terminal sequences of the C/Bristol VP4 gene were used to amplify the corresponding VP4 gene of a human group C rotavirus from Belém, Brazil. Nucleotide sequence comparisons of the Bristol and Belém VP4 genes revealed 45 differences of which only 6 were predicted to give amino acid changes Alignment of the two human VP4 sequences with the prototype porcine group C/Cowden rotavirus VP4 showed only 71.2% nucleotide sequence identity. Protein sequence alignments showed that the human group C rotavirus VP4 sequences were 8 amino acids longer than the porcine VP4 sequence with an insertion of 6 amino acids, 252NSKLGD 257 adjacent to the proposed proteolytic cleavage region (amino acids 231-250). The large overall number of differences between the human and porcine VP4 sequences strongly suggested that the porcine C/Cowden isolate may belong to a different group C rotavirus P "serotype." In contrast the very close similarity of the VP4 sequences of the UK and Brazilian group C rotaviruses support the hypothesis that these human isolates originate from a recent common ancestor.