Identification and characterization of the bovine herpesvirus 5 US4 gene and gene products

The BHV-5 strain N569 (BHV-5/N569) homolog to the BHV-1 US4 gene was sequenced and characterized. RNA analyses showed that a 1.8-kb mRNA which contains the BHV-5/N569 US4 open reading frame initiates 55 nucleotides upstream from the predicted translational start codon and terminates 17 nucleotides downstream from the consensus sequence for polyadenylation. Comparison of the deduced amino acid sequences of the predicted US4 encoded proteins of BHV-5/N569 and BHV-1 strain Schönböken (BHV-1/Schö) revealed 75% identity. An antiserum, raised in rabbits after infection with a BHV-5/N569 US4 ORF expressing recombinant vaccinia virus, specifically precipitated a 65-kDa protein and a diffusely migrating protein species with an apparent molecular mass between 90 and >240 kDa from the supernatant of BHV-5/N569 infected cells. Treatment of immmunprecipitated proteins with chondroitinase AC demonstrated that the latter contains glycosaminoglycans. The mobility of the BHV-5/N569 US4 gene products was identical to the BHV-1 US4 ORF encoded glycoprotein G (gG) and glycoproteoglycan G (gpgG; G. M. Keil, T. Engelhardt, A. Karger, and M. Enz,J. Virol.70, 3032–3038, 1996) and were therefore named BHV-5 gG and BHV-5 gpgG. Immunoprecipitations with sera from BHV-1 infected cattle indicated a type-specific immune response to gG, since these sera failed to react with vaccinia virus-expressed gG-5 but recognized vaccinia virus-expressed gG-1.