Characterization of the proteinase specified by varicella-zoster virus gene 33.

McMillan, David; Kay, John; Mills, John

doi:10.1099/0022-1317-78-9-2153

Cited by 9 publications

(6 citation statements)

References 18 publications

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“…This contrasts with the findings reported for the proteinases from the ␣-herpesviruses, e.g. HSV-1 and VZV (11,12), which appear to require much longer synthetic peptides, e.g. spanning P 3 to P 7 Ј for the R-site and P 5 to P 8 Ј for the M-site, for efficient substrate cleavage (11).…”

Section: Resultscontrasting

confidence: 80%

“…From work characterizing HCMV proteinase (19,20), it would appear that HHV-6 proteinase resembles this enzyme rather more closely than HSV-1, HSV-2, or VZV proteinases (11,12). The equivalent residue in HCMV proteinase to Tyr 227 in the HHV-6 sequence is Tyr 253 , which is also positioned at the P 4 location relative to the C-terminal Ala residue generated upon R-site cleavage.…”

Section: 20 10mentioning

confidence: 99%

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Autoprocessing and Peptide Substrates for Human Herpesvirus 6 Proteinase

Tigue

Kay

1998

Journal of Biological Chemistry

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Autoprocessing of the precursor form of human herpesvirus 6 (HHV-6) proteinase at two sites (termed M and R) is required to generate the mature enzyme. Kinetic constants were determined for the hydrolysis of a series of synthetic peptide substrates by mature HHV-6 proteinase, purified to homogeneity. Truncation or replacement of individual residues in peptides mimicking the R-site sequence, indicated that the minimum length for effective hydrolysis by the viral enzyme was P 4 -P 3 -P 2 -Ala*Ser-P 2 -P 3 -P 4 and revealed the importance of the P 1 Ala and P 4 Tyr residues. Consequently, relevant (P 1 or P 4 ) mutations were introduced into the precursor form of the proteinase and the ability of these altered proteins to autoprocess was examined. Introduction of Val in place of the P 1 Ala at the M-site essentially abrogated cleavage but mature HHV-6 proteinase was still generated by cleavage at the R-site, indicating that processing of the M-site is not a prerequisite for cleavage of the R-site in the precursor. At the R-site, mutation of the P 1 Ala, or of the preceding P 4 Tyr residue, prevented processing at the R-site in the precursor so that the mature form of HHV-6 proteinase was not generated. The accumulated data suggest a possible new approach to the design of inhibitors for therapeutic intervention in the life cycle of herpesviruses.Eight herpesviruses have been implicated to date in the etiology of a number of human diseases. These are subdivided into three families on the basis of their biological properties, host cell range and the organization of their genomes (1). Herpes simplex virus type 1 (HSV-1) 1 and type 2 (HSV-2) and varicella zoster virus (VZV), the causative agent of chicken pox and shingles, comprise the ␣-subgroup while Epstein-Barr virus and the recently discovered human herpesvirus 8 (which has been implicated in Kaposi's sarcoma in human immunodeficiency virus-positive individuals; Ref. 2) constitute the ␥-subfamily. The ␤-subgroup consists of human cytomegalovirus (HCMV) and human herpesviruses 6 and 7 (HHV-6 and HHV-7). HHV-6 causes the childhood illness exanthem subitum (3) and has also been implicated in the progression of a number of other diseases, including multiple sclerosis (4) and chronic fatigue syndrome (5).In common with the other herpesviruses, HHV-6 encodes its own proteinase, which is essential for capsid maturation, DNA packaging, and the ultimate formation of new virus particles (6). The mature proteinase consists of 230 residues but is synthesized in the form of a precursor, which has an additional 298 residues attached to the C terminus of the mature enzyme (see Ref. 7 and Fig. 1). Autolytic removal of these residues (which themselves constitute a form of the viral assembly protein; Ref. 6) releases the N-terminal proteinase in its mature form; processing takes place at two locations positioned, respectively, at the C terminus of the proteinase (R-site; Fig. 1) and 37 residues from the C terminus of the precursor (M-site; Fig. 1). In the HHV-6 precursor, just as ...

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Section: Resultscontrasting

confidence: 80%

Section: 20 10mentioning

confidence: 99%

Autoprocessing and Peptide Substrates for Human Herpesvirus 6 Proteinase

Tigue

Kay

1998

Journal of Biological Chemistry

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“…VZV and SVV have the smallest genomes of the herpesvirus family. VZV encodes 68 ORFs, 3 novel ORFs and 3 ORF duplications (Cohen et al , 2007; Kemble et al , 2000; McMillan et al , 1997; Ross et al , 1997). Similarly, SVV encodes 72 ORFs with 69 unique ORFs and 3 ORFs duplicated in the terminal repeat sequence (TRS).…”

Section: Discussionmentioning

confidence: 99%

Simian varicella virus gene expression during acute and latent infection of rhesus macaques

et al. 2011

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Varicella zoster virus (VZV) is a neurotropic α-herpesvirus that causes chickenpox during primary infection and establishes latency in sensory ganglia. Reactivation of VZV results in herpes zoster and other neurological complications. Our understanding of the VZV transcriptome during acute and latent infection in immune competent individuals remains incomplete. Infection of rhesus macaques with the homologous simian varicella virus (SVV) recapitulates the hallmarks of VZV infection. We therefore characterized the SVV transcriptome by quantitative real-time reverse transcriptase PCR (RT-qPCR) during acute infection in bronchial alveolar lavage (BAL) cells and peripheral blood mononuclear cells (PBMC), and during latency in sensory ganglia obtained from the same rhesus macaques. During acute infection, all known SVV open reading frames (ORFs) were detected and the most abundantly expressed ORFs are involved in virus replication and assembly such as the transcriptional activator ORF 63 and the structural proteins ORF 41 and ORF 49. In contrast, latent SVV gene expression is highly restricted. ORF 61, a viral transactivator and latency-associated transcript, is the most prevalent transcript detected in sensory ganglia. We also detected ORFs A, B, 4, 10, 63, 64, 65, 66 and 68 though significantly less frequently than ORF 61. This comprehensive analysis has revealed genes that potentially play a role in the establishment and/or maintenance of SVV latency.

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“…ORFs 33 and 33.5 encode highly immunogenic proteins associated with the assembly of virus nucleocapsids (12,16,27,33). The ORF 33 transcript encodes the full-length precursor protein, which is posttranslationally cleaved to yield the assembly protein protease (ORF 33) and the structural protein substrate (ORF 33.5).…”

Section: Discussionmentioning

confidence: 99%

Array Analysis of Viral Gene Transcription during Lytic Infection of Cells in Tissue Culture with Varicella-Zoster Virus

Cohrs¹,

Hurley²,

Gilden

2003

J Virol

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Varicella-zoster virus (VZV), a neurotropic alphaherpesvirus, causes childhood chickenpox (varicella), becomes latent in dorsal root and autonomic ganglia, and reactivates decades later to cause shingles (zoster) and other neurologic complications. Although the sequence and configuration of VZV DNA have been determined, relatively little is known about viral gene expression in productively infected cells. This is in part because VZV is highly cell associated, and sufficient titers of cell-free virus for use in synchronizing infection do not develop. PCR-based transcriptional arrays were constructed to simultaneously determine the relative abundance of the Ϸ70 predicted VZV open reading frames (ORFs). Fragments (250 to 600 bp) from the 5 and 3 end of each ORF were PCR amplified and inserted into plasmid vectors. The virus DNA inserts were amplified, quantitated, and spotted onto nylon membranes. Probing the arrays with radiolabeled cDNA synthesized from VZV-infected cells revealed an increase in the magnitude of the expressed VZV genes from days 1 to 3 after low-multiplicity virus infection but little change in their relative abundance. The most abundant VZV transcripts mapped to ORFs 9/9A, 64, 33/33A, and 49, of which only ORF 9 corresponded to a previously identified structural gene. Array analysis also mapped transcripts to three large intergenic regions previously thought to be transcriptionally silent, results subsequently confirmed by Northern blot and reverse transcription-PCR analysis. Array analysis provides a formidable tool to analyze transcription of an important ubiquitous human pathogen. Varicella-zoster virus (VZV) is propagated by cocultivating infected cells with uninfected cells.Because VZV is cell associated and high titers of cell-free virus that can be used to synchronize infection do not develop, analysis of VZV gene transcription in infected cells in tissue culture has been limited. Three reports described global analysis of VZV transcription in cultures harvested at advanced stages of infection; Northern blot analysis of RNA extracted from late-stage VZV-infected cultures identified 58 to 67 discrete transcripts mapping throughout the virus genome (26, 32), while single-stranded DNA probes used in Northern blots revealed the direction of transcription of 57 of the 58 previously identified VZV transcripts and also identified 20 novel virus transcripts (35). Because the RNA in those studies was sampled at the height of cytopathic effect, the transcripts were presumed to reflect VZV structural genes.To better understand the pattern of virus gene transcription during lytic infection, we constructed PCR-based VZV transcriptional arrays to analyze the magnitude of expression and relative abundance of each predicted VZV gene as well as detect transcription for three large intergenic regions of the virus genome. MATERIALS AND METHODSVirus and cells. VZV (strain Ellen) was propagated as described previously (13) by cocultivation of infected and uninfected BSC-1 cells, a continuous cell line derived...

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Characterization of the proteinase specified by varicella-zoster virus gene 33.

Cited by 9 publications

References 18 publications

Autoprocessing and Peptide Substrates for Human Herpesvirus 6 Proteinase

Autoprocessing and Peptide Substrates for Human Herpesvirus 6 Proteinase

Simian varicella virus gene expression during acute and latent infection of rhesus macaques

Array Analysis of Viral Gene Transcription during Lytic Infection of Cells in Tissue Culture with Varicella-Zoster Virus

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