Sequence and genetic arrangement of the U L region of the monkey B virus ( Cercopithecine herpesvirus 1) genome and comparison with the U L region of other primate herpesviruses

Ohsawa, Kazutaka; Black, Darla H.; Sato, Hiroshi; Rogers, Kristin M.; Eberle, R.

doi:10.1007/s00705-003-0011-2

Cited by 23 publications

(18 citation statements)

References 39 publications

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“…The sequences of the cloned fragments were identical to the genomic sequences of the corresponding B virus genes (42), with the exception of the mgG fragment, as there was a 132-bp deletion (positions 1096 to 1227 in the gG gene) in the constructed mgG donor plasmid. This deleted DNA fragment, which encodes a PAPTTT amino acid repeat, is present in the gG sequence of the E2490 strain but is not found in any of the other sequenced gG genes of B virus clinical isolates (40). Since the deletion construct encodes an mgG protein that actually mimics those produced during natural infections, we decided to use this construct for production of the mgG recombinant baculovirus.…”

Section: Resultsmentioning

confidence: 99%

Production of Herpes B Virus Recombinant Glycoproteins and Evaluation of Their Diagnostic Potential

et al. 2005

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B virus (cercopithecine herpesvirus 1) is the only deadly alphaherpesvirus that is zoonotically transmissible from macaques to humans. The detection of humoral immune responses is the method of choice for the rapid identification of B virus-infected animals. We evaluated the diagnostic potential of recombinant B virus glycoproteins for the detection of immunoglobulin G (IgG) antibodies in monkey and human sera. Glycoproteins B, C, and E and secreted (sgG) and membrane-associated (mgG) segments of glycoprotein G (gG) were expressed in the baculovirus expression system, while gD was expressed in CHO cells. We developed recombinant protein-based IgG enzyme-linked immunosorbent assays (ELISAs) and compared their diagnostic efficacies by using B virus antibody-negative (n ‫؍‬ 40) and -positive (n ‫؍‬ 75) macaque sera identified by a whole antigenbased ELISA and Western blotting. The diagnostic sensitivities of the gB-, gC-, gD-, and mgG-ELISAs were 100, 97.3, 88.0, and 80.0%, respectively. The specificities of the gB-, gC-, and gD-ELISAs and of the mgG-ELISA were 100 and 97.5%, respectively. In contrast, the sensitivities and specificities of sgG-and gE-ELISAs were low, suggesting that sgG and gE are less effective diagnostic antigens. Sera from nonmacaque monkeys crossreacted with gB, gC, and gD, and only baboon sera reacted weakly with mgG. Human herpes simplex virus type 1 (HSV-1)-and HSV-2-positive sera pools reacted with gB and gD, whereas sera from B virus-infected individuals reacted with all four antigens. These data indicate that gB, gC, gD, and mgG have a high diagnostic potential for B virus serodiagnosis in macaques, whereas mgG may be a valuable antigen for discrimination between antibodies induced by B virus and those induced by other, closely related alphaherpesviruses, including HSV-1 and -2.Human infection with B virus (also called cercopithecine herpesvirus 1, monkey B virus, and herpes B virus) is the most feared occupational hazard among individuals working with macaque monkeys, since fatality is often the outcome of infection, which proceeds in the absence of effective antiviral therapy (25,56). The use of macaques in research has been steadily growing over the last decade and is expected to rise quickly in the near future due to the increasing demands for these animals for use in HIV/AIDS investigations, vaccine trials, drug testing, and research into bioterrorism agents. As macaque usage increases, frequencies of human exposures to B virus are increasing as well. Rapid and accurate diagnostic tests are urgently needed to aid in the early identification of clinical cases, which is essential for a timely initiation of antiviral therapies in zoonotically infected humans. In addition to human diagnostics, enhanced assays are required for monitoring specific-pathogen-free (SPF) macaque colonies established by the National Institutes of Health for the breeding of B virusfree animals (55), as these animals often demonstrate only very low levels of antibody.Unfortunately, a direct diagnosis of infectio...

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Section: Resultsmentioning

confidence: 99%

Production of Herpes B Virus Recombinant Glycoproteins and Evaluation of Their Diagnostic Potential

et al. 2005

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“…Sequence analysis of the partial U S regions of B virus and simian agent 8 virus demonstrated that human and primate viruses are colinear in this genomic segment (51). However, only a limited number of B virus gene sequences from the U L region have been published (3,36,62,63), and nothing has been reported about the structure and the gene content of the repeat genomic elements.…”

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confidence: 99%

Complete Sequence and Comparative Analysis of the Genome of Herpes B Virus ( Cercopithecine Herpesvirus 1 ) from a Rhesus Monkey

Perelygina

Zhu

Zurkuhlen

et al. 2003

J Virol

101

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The complete DNA sequence of herpes B virus (Cercopithecine herpesvirus 1) strain E2490, isolated from a rhesus macaque, was determined. The total genome length is 156,789 bp, with 74.5% G؉C composition and overall genome organization characteristic of alphaherpesviruses. The first and last residues of the genome were defined by sequencing the cloned genomic termini. There were six origins of DNA replication in the genome due to tandem duplication of both oriL and oriS regions. Seventy-four genes were identified, and sequence homology to proteins known in herpes simplex viruses (HSVs) was observed in all cases but one. The degree of amino acid identity between B virus and HSV proteins ranged from 26.6% (US5) to 87.7% (US15). Unexpectedly, B virus lacked a homolog of the HSV ␥ 1 34.5 gene, which encodes a neurovirulence factor. Absence of this gene was verified in two low-passage clinical isolates derived from a rhesus macaque and a zoonotically infected human. This finding suggests that B virus most likely utilizes mechanisms distinct from those of HSV to sustain efficient replication in neuronal cells. Despite the considerable differences in G؉C content of the macaque and B virus genes (51% and 74.2%, respectively), codons used by B virus are optimal for the tRNA population of macaque cells. Complete sequence of the B virus genome will certainly facilitate identification of the genetic basis and possible molecular mechanisms of enhanced B virus neurovirulence in humans, which results in an 80% mortality rate following zoonotic infection.Comparative genome analyses of closely related viruses offer insight into the protein coding capacities of viral genomes (18, 69), a means to biologically classify viruses (2, 19) and identify viral genes involved in virulence and pathogenicity (1,37,67). The number of completely sequenced viral genomes has been increasing rapidly and now exceeds 1,000, including 29 herpesvirus genomes (GenBank data, http://www.ncbi.nlm.nih.gov /PMGifs/Genomes/viruses.html).B virus (Cercopithecine herpesvirus 1, monkey B virus) is a member of the subfamily Alphaherpesvirinae, which together with human herpes simplex virus types 1 and 2 (HSV-1 and HSV-2) constitutes the genus Simplexvirus. B virus generally causes only mild localized or asymptomatic infections in its natural hosts, Asian monkeys of the genus Macaca (33,34,74). In contrast, B virus infections in foreign hosts, humans or monkey species other than macaques, often result in encephalitis, encephalomyelitis, and death (53,73,74).The genome organization of B virus is similar to that of HSV-1 and HSV-2: the unique long (U L ) and unique short (U S ) segments flanked by inverted long (R L ) and short (R S ) repeat sequences are covalently joined in four possible isomeric configurations (26). Sequence analysis of the partial U S regions of B virus and simian agent 8 virus demonstrated that human and primate viruses are colinear in this genomic segment (51). However, only a limited number of B virus gene sequences from the U L region have b...

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“…It is important to recognize that the RXP-containing proteins are otherwise unrelated, as SM, in fact, shows structural and functional homology to an additional alphaherpesvirus protein, the herpes simplex virus ICP27 polypeptide (2,3,8,13,18,42,45). Similar transactivator proteins encoded by other herpesviruses do not contain an RXP repetitive element, and Us11-homologous proteins are found only in HSV-1, HSV-2, and the simian B virus (16,31,35). Importantly, however, several human herpesviruses are known to encode activities that can regulate PKR activation regardless of whether they produce an RXPcontaining protein (5,10).…”

Section: Discussionmentioning

confidence: 99%

“…After rocking for 1 h at room temperature, the beads were collected by centrifugation, suspended in 0.1 ml of SMPB plus 3% fraction V bovine serum albumin, 0.5% Triton X-100, and 1 mM CaCl 2 and treated for 20 min at room temperature with 8 U of micrococcal nuclease per ml or 8 U of micrococcal nuclease, 50 g of RNase A, and 300 U of RNase T 1 per ml. 35 S-labeled PKR produced by translation in vitro according to the manufacturer's directions (wheat germ T7 TNT quick coupled transcription-translation system; Promega, Madison, Wis.) was likewise treated with nuclease at the conclusion of the synthesis reaction. Phenylmethylsulfonyl fluoride was then added to both the beads loaded with GST fusion protein and the nucleasetreated in vitro translation reaction to a final concentration of 100 M. Then 1 l of the in vitro-translated lysate was added to the nuclease-treated beads containing bound fusion protein, and the final concentration of phenylmethylsulfonyl fluoride was brought to 200 M. Binding reaction mixtures were incubated at 30°C for 30 min and agitated manually every 5 min.…”

Section: Methodsmentioning

confidence: 99%

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Identification of a Lytic-Cycle Epstein-Barr Virus Gene Product That Can Regulate PKR Activation

Poppers

Mulvey

Perez

et al. 2003

J Virol

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The Epstein-Barr virus (EBV) SM protein is a posttranscriptional regulator of viral gene expression. Like many transactivators encoded by herpesviruses, SM transports predominantly unspliced viral mRNA cargo from the nucleus to the cytosol, where it is subsequently translated. This activity likely involves a region of the protein that has homology to the herpes simplex virus type 1 (HSV-1) ICP27 gene product, the first member of this class of regulators to be discovered. However, SM also contains a repetitive segment rich in arginine and proline residues that is dispensable for its effects on RNA transport and splicing. This portion of SM, comprised of RXP triplet repeats, shows homology to the carboxyl-terminal domain of Us11, a double-stranded RNA (dsRNA) binding protein encoded by HSV-1 that inhibits activation of the cellular PKR kinase. To evaluate the intrinsic ability of SM to regulate PKR, we expressed and purified several SM protein derivatives and examined their activity in a variety of biochemical assays. The full-length SM protein bound dsRNA, associated physically with PKR, and prevented PKR activation. Removal of the 37-residue RXP domain significantly compromised all of these activities. Furthermore, the SM RXP domain was itself sufficient to inhibit PKR activation and interact with the kinase. Relative to its Us11 counterpart, the SM RXP segment bound dsRNA with reduced affinity and responded differently to single-stranded competitor polynucleotides. Thus, SM represents the first EBV gene product expressed during the lytic cycle that can prevent PKR activation. In addition, the RXP repeat segment appears to be a conserved herpesvirus motif capable of associating with dsRNA and modulating activation of the PKR kinase, a molecule important for the control of translation and the cellular antiviral response.Early in the course of a lytic infection, Epstein-Barr virus (EBV) expresses a polypeptide 479 amino acids in length known as SM, Mta, or EB2 that regulates viral gene expression posttranscriptionally (9, 13, 29). The SM polypeptide, encoded by a spliced mRNA that joins the BSLF2 and BMLF1 open reading frames, can transactivate reporter gene expression in a promoter-independent, gene-specific fashion in transienttransfection assays (4,9,25,29,41,42). In addition, SM has been reported to bind RNA, shuttle into and out of the nucleus, inhibit expression of intron-containing genes, and activate expression of genes that lack introns (2,3,8,13,18,41,42,45). Many EBV genes important for viral replication in productively infected cells do not contain introns, and the cytoplasmic accumulation of their mRNAs is regulated by SM, as are cytoplasmic levels of mRNAs transcribed from reporter genes that do not contain introns (13,26,45). Other studies, however, provide evidence that SM inhibits splicing of precursors containing weak 5Ј splice site consensus sequences and mediates their accumulation in the cytosol (3,18,45). Finally, SM associates with the cellular SC35 splicing factor (8,26,45

show abstract

Sequence and genetic arrangement of the U L region of the monkey B virus ( Cercopithecine herpesvirus 1) genome and comparison with the U L region of other primate herpesviruses

Cited by 23 publications

References 39 publications

Production of Herpes B Virus Recombinant Glycoproteins and Evaluation of Their Diagnostic Potential

Production of Herpes B Virus Recombinant Glycoproteins and Evaluation of Their Diagnostic Potential

Complete Sequence and Comparative Analysis of the Genome of Herpes B Virus ( Cercopithecine Herpesvirus 1 ) from a Rhesus Monkey

Identification of a Lytic-Cycle Epstein-Barr Virus Gene Product That Can Regulate PKR Activation

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