Complete nucleotide sequence of the herpesvirus simiae glycoprotein G gene and its expression as an immunogenic fusion protein in bacteria

Abstract

The nucleotide sequence of a 2384 bp portion within the unique short (Us) region of the herpesvirus simiae (simian herpes B virus; SHBV) genome is presented. A partial and a complete open reading frame (ORF) were found within this nucleotide sequence. The partial ORF encodes the C terminus (147 amino acids) of a protein kinase which is highly conserved in the herpes simplex virus type 1 (HSV-1) and type 2 (HSV-2) and simian agent 8 (SA8) U s regions. The complete ORF is located 3' to the partial ORF within the 2384 bp sequence and encodes a 593 amino acid g|ycoprotein which appears to be closely related to the SA8 glycoprotein G (gG), but shares little amino acid similarity with gG of HSV-1 and -2. However, the complete ORF shares certain features conserved among most alphaherpesvirus gGs, notably three highly conserved cysteine residues and an adjacent N-glycosylation site. Therefore, it was concluded that this complete ORF encodes the SHBV gG. The 358 amino acid C-terminal portion of SHBV gG was expressed in Escherichia coli as a fusion protein and this was detected by immunoblotting with sera from cynomolgus monkeys which were either experimentally or naturally infected with SHBV. The purified fusion protein was inoculated into rabbits to raise an antiserum which recognized a number of apparently SHBV gGspecific protein bands in extracts from SHBV-infected simian cells.

Equine herpesvirus-4 glycoprotein G is secreted as a disulphide-linked homodimer and is present as two homodimeric species in the virion.

“…Specifically, HSV-2, pseudorabies virus, herpesvirus simiae, BHV-1, BHV-5 and ILTV gG homologues have not been shown to be integral components of the viral envelope although gG-related species have been identified in infected radiolabelled cells (Balachandran & HuttFletcher, 1985 ;Bennett et al, 1986 ;Keil et al, 1996 ;Kongsuwan et al, 1993 ;Slomka et al, 1995).…”

“…Of these, only four, gB, gD and the gH\gL complex, are essential for replication in vitro in cell culture, although it is presumed that the remainder serve important roles in vivo in viral pathogenesis. Glycoprotein G (gG) homologues have been identified in most alphaherpesviruses that have been examined (Colle et al, 1992 ;Crabb et al, 1992 ;Eberle et al, 1993 ;Ficorilli et al, 1995 ;Fukushi et al, 1997 ;Keil et al, 1996 ;Kongsuwan et al, 1993 ;McGeoch et al, 1987 ;Rea et al, 1985 ;Slomka et al, Fax j613 9416 2616. e-mail heidid!ariel.its.unimelb.edu.au associated species, gGV L and gGV S , are present in the virion as 140 and 20 kDa proteins, respectively. As gGS and gGV L do not appear to associate stably with other viral proteins, it is most likely that each species exists as a disulphide-linked homodimer.…”

Sequence and Genetic Arrangement of the USRegion of the Monkey B Virus (Cercopithecine Herpesvirus 1) Genome and Comparison with the USRegions of Other Primate Herpesviruses

“…The US4 ORF encodes a glycoprotein designated gG (27), which varies considerably among these viruses. All of the simian virus gGs are more similar to the gG of HSV-2 than to that of HSV-1, having a hydrophobic Nterminal signal sequence, two distinct regions of the extracellular domain separated by a proteolytic cleavage site, a transmembrane domain, and a highly charged cytoplasmic tail (27,36,34). The more N-terminal of the extracellular domains is relatively conserved in sequence, with six Cys residues and one N-linked glycosylation site being positionally conserved.…”

“…When transmitted to nonmacaque primates, BV produces severe infections which usually involve the central nervous system and are frequently fatal (31,37,38). The molecular basis for the extreme neuropathology of BV in nonmacaque species is an intriguing question that remains unanswered.While sequences for a number of genes of BV have been reported (3,9,23,34,35,37), genomic characterization of BV has largely been limited to restriction analysis and gene mapping by hybridization with HSV gene probes (10,16,17). Such studies have suggested that, for the most part, the BV genome is colinear with that of HSV.…”

“…While sequences for a number of genes of BV have been reported (3,9,23,34,35,37), genomic characterization of BV has largely been limited to restriction analysis and gene mapping by hybridization with HSV gene probes (10,16,17). Such studies have suggested that, for the most part, the BV genome is colinear with that of HSV.…”