Molecular Characterization of HE, M, and E Genes of Winter Dysentery Bovine Coronavirus Circulated in Korea During 2002–2003

The different bovine coronavirus (BCoV) strains or isolates exhibited various degrees of substitutions, resulting in altered antigenicity and pathogenicity of the virus. In the previous our study, we demonstrated that the spike glycoprotein gene of Korean winter dysentery (WD) BCoV had a genetic property of both enteric (EBCV) and respiratory BCoV (RBCV) and were significantly distinct from the ancestral enteric strains. In the present study, therefore, we analyzed the other structure genes, the hemagglutinin/esterase (HE) protein, the transmembrane (M) protein and the small membrane (E) protein to characterize 10 WD BCoV circulated in Korea during 2002–2003 and compared the nucleotide and deduced amino acid sequences with the other known BCoV. Phylogenetic analysis indicated that the HE gene among BCoV could be divided into three groups. The first group included only RBCV, while the second group contained calf diarrhea BCoV, RBCV, WD and EBCV, respectively. The third group possessed only all Korean WD strains which were more homologous to each other and were sharply distinct from the other known BCoV, suggesting Korean WD strains had evolutionary distinct pathway. In contrast, the relative conservation of the M and E proteins of BCoV including Korean WD strains and the other coronaviruses suggested that structural constraints on these proteins are rigid, resulting in more limited evolution of these proteins. In addition, BCoV and human coronavirus HCV-OC43 contained four potential O-glycosylation sites in the M gene. However, the M gene sequence of both BCoV and HCV-OC43 might not contain a signal peptide, suggesting the M protein might be unlikely to be exposed to the O-glycosylation machinery in vivo.