Characterization of the herpes simplex virus type 2 (HSV-2) US2 gene product and a US2-deficient HSV-2 mutant.

The sequence of the unique short (U S ) region of monkey B virus (BV) was determined. The 13 genes identified are arranged in the same order and orientation as in herpes simplex virus (HSV). These results demonstrate that the BV U S region is entirely colinear with that of HSV type 1 (HSV-1), HSV-2, and simian agent 8 virus.Cercopithecine herpesvirus 1 (monkey B virus [BV]) is a member of the alphaherpesvirus subfamily indigenous in Asiatic macaque monkeys. The natural history of BV in macaques is similar to that of the human herpes simplex virus types 1 and 2 (HSV-1 and HSV-2) in humans, of simian agent 8 virus (SA8) in green monkeys, and of Herpesvirus papio 2 (HVP-2) in baboons (11,12,38). 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. However, it has been reported that the unique short (U S ) region of the BV genome may not be colinear with that of HSV (16). Here we present the sequence of the U S region of the BV genome and its genetic layout relative to that of HSV and SA8.The DNA sequence of the BV rhesus genotype (BVrh) strain E2490 (35) U S region was determined by a combination of cloning restriction fragments of the genome and PCR amplification of small gaps in the sequence with genomic BV DNA as a template. The BVrh U S region (GenBank accession no. AB 077432) is 14,447 nucleotides long, slightly longer than the 12,980 bp reported for HSV-1 strain 17 (26) but close to the 14,329-bp HSV-2 U S region (6). The BVrh U S sequence presented here does not include sequences aligning with the N-terminal 2 codons of the US12 open reading frame (ORF). The overall GϩC content of the BVrh U S region is 73.2%, close to the 75% GϩC estimated for the entire BV genome (19). The coding sequences in the U S region have a combined GϩC content of 74.4%, a value somewhat higher than that for the noncoding intergenic regions (68.9%). BV exhibits a strong bias toward use of codons with G or C in the third position (89.6%) and Arg and Leu codons with C in the first position (93.4%). While this GC bias is strong, it is not as extreme as in the SA8 U S region (76.8% GϩC overall, 92.9% GC in the third position, 97.2% C in the first position of the Arg and Leu codons [13]).Analysis of the BVrh U S sequence for ORFs, homology of predicted amino acid sequences with HSV polypeptides, promoter and transcriptional initiation sites, and mRNA termination sites [consensus poly(A) signals associated with mRNA termination motifs] indicates that the genetic layout of the BVrh U S regio...

supporting

confidence: 75%

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

Ohsawa

Black

Sato

et al. 2002

“…The US2 homologue in HSV-1 and HSV-2 seems dispensable in tissue culture and is not involved in HSV-2 neuropathogenesis in mice (52,61). BHV-5 US4 encodes a glycosaminoglycan-associated protein (31) and is 72% identical to BHV-1 gG, which is involved in cell-cell virus transmission in vitro (69) and in prevention of apoptosis in certain cell lines (68).…”

Section: Resultsmentioning

confidence: 99%

Genome of Bovine Herpesvirus 5

Delhon

Moraes

et al. 2003

143

114

Here we present the complete genomic sequence of bovine herpesvirus 5 (BHV-5), an alphaherpesvirus responsible for fatal meningoencephalitis in cattle. The 138,390-bp genome encodes 70 putative proteins and resembles the ␣2 subgroup of herpesviruses in genomic organization and gene content. BHV-5 is very similar to BHV-1, the etiological agent of infectious bovine rhinotracheitis, as reflected by the high level of amino acid identity in their protein repertoires (average, 82%). The highest similarity to BHV-1 products (>95% amino acid identity) is found in proteins involved in viral DNA replication and processing (UL5, UL15, UL29, and UL39) and in virion proteins (UL14, UL19, UL48, and US6). Among the least conserved (<75%) are the homologues of immediate-early (IE) proteins BICP0, BICP4, and BICP22, the three proteins being longer in BHV-5 than in BHV-1. The structure of the BHV-5 latency-related (LR) region departs markedly from that of BHV-1 in both coding and transcriptional regulatory regions. Given the potential significance of IE genes and the LR region in virus-neuron interactions, it is likely these differences contribute to BHV-5 neuropathogenicity.

“…The Us2 homologs from both EHV-1 and HSV-2 are incorporated into mature virions (29,48). To determine whether PRV Us2 was packaged into virions, we subjected purified extracellular virions to a protease protection assay described previously (7,43).…”

Section: Identification Of Us2 Protein In Prv-infected Cell Lysatesmentioning

confidence: 99%

“…EHV-1 Us2-null mutants are attenuated in mice after intranasal inoculation (48). In contrast, HSV-2 Us2-null mutants were as virulent as wild-type virus in mice after footpad inoculation (29) or intravaginal infection (26). EHV-1 Us2-null mutants formed small plaques on monolayers of rabbit kidney cells despite having normal kinetics in single-step growth studies (48).…”

mentioning

confidence: 99%

“…Despite this conservation, no function has been assigned and, when tested, the protein is not required for virus growth in cultured cells. The herpes simplex virus type 2 (HSV-2) (23,26,29), HSV-1 (42,47,71), and equine herpesvirus type 1 (EHV-1) (48) homologs are virion structural proteins. Late after infection of Vero cells with HSV-2, the Us2 protein localizes to the nucleus and cytoplasm (29).…”

mentioning

confidence: 99%

See 1 more Smart Citation

The Pseudorabies Virus Us2 Protein, a Virion Tegument Component, Is Prenylated in Infected Cells

Clase

Lyman

Rio

et al. 2003

The Us2 gene is conserved among alphaherpesviruses, but its function is not known. We demonstrate here that the pseudorabies virus (PRV) Us2 protein is synthesized early after infection and localizes to cytoplasmic vesicles and to the plasma membrane, despite the lack of a recognizable signal sequence or membranespanning domain. Us2 protein is also packaged as part of the tegument of mature virions. The Us2 carboxyterminal four amino acids comprise a CAAX motif, a well-characterized signal for protein prenylation. Treatment of infected cells with lovastatin, a drug that disrupts protein prenylation, changed the relative electrophoretic mobility of Us2 in sodium dodecyl sulfate-polyacrylamide gels. In addition, lovastatin treatment caused a dramatic relocalization of Us2 to cytoplasmic punctate structures associated with microtubules, which appeared to concentrate over the microtubule organizing center. When the CAAX motif was changed to GAAX and the mutant protein was synthesized from an expression plasmid, it concentrated in punctate cytoplasmic structures reminiscent of Us2 localization in infected cells treated with lovastatin. We suggest that prenylation of PRV Us2 protein is required for proper membrane association. Curiously, the Us2 protein isolated from purified virions does not appear to be prenylated. This is the first report to describe the prenylation of an alphaherpesvirus protein.