Abstract:
:The varicella-zoster virus (VZV) open reading frame 54 (ORF54) gene encodes an 87-kDa monomer that oligomerizes to form the VZV portal protein, pORF54. pORF54 was hypothesized to perform a function similar to that of a previously described herpes simplex virus 1 (HSV-1) homolog, pUL6. pUL6 and the associated viral terminase are required for processing of concatemeric viral DNA and packaging of individual viral genomes into preformed capsids. In this report, we describe two VZV bacterial artificial chromosome (BAC) constructs with ORF54 gene deletions, Δ54L (full ORF deletion) and Δ54S (partial internal deletion). The full deletion of ORF54 likely disrupted essential adjacent genes (ORF53 and ORF55) and therefore could not be complemented on an ORF54-expressing cell line (ARPE54). In contrast, Δ54S was successfully propagated in ARPE54 cells but failed to replicate in parental, noncomplementing ARPE19 cells. Transmission electron microscopy confirmed the presence of only empty VZV capsids in Δ54S-infected ARPE19 cell nuclei. Similar to the HSV-1 genome, the VZV genome is composed of a unique long region (UL) and a unique short region (US) flanked by inverted repeats. DNA from cells infected with parental VZV (VZVLUC strain) contained the predicted UL and US termini, whereas cells infected with Δ54S contained neither. This result demonstrates that Δ54S is not able to process and package viral DNA, thus making pORF54 an excellent chemotherapeutic target. In addition, the utility of BAC constructs Δ54L and Δ54S as tools for the isolation of site-directed ORF54 mutants was demonstrated by recombineering single-nucleotide changes within ORF54 that conferred resistance to VZV-specific portal protein inhibitors. Importance: Antivirals with novel mechanisms of action would provide additional therapeutic options to treat human herpesvirus infections. Proteins involved in the herpesviral DNA encapsidation process have become promising antiviral targets. Previously, we described a series of N-α-methylbenzyl-N'-aryl thiourea analogs that target the VZV portal protein (pORF54) and prevent viral replication in vitro. To better understand the mechanism of action of these compounds, it is important to define the structural and functional characteristics of the VZV portal protein. In contrast to HSV, no VZV mutants have been described for any of the seven essential DNA encapsidation genes. The VZV ORF54 deletion mutant described in this study represents the first VZV encapsidation mutant reported to date. We demonstrate that the deletion mutant can serve as a platform for the isolation of portal mutants via recombineering and provide a strategy for more in-depth studies of VZV portal structure and function.
journal_name
J Viroljournal_title
Journal of virologyauthors
Visalli MA,House BL,Selariu A,Zhu H,Visalli RJdoi
10.1128/JVI.00376-14subject
Has Abstractpub_date
2014-07-01 00:00:00pages
7973-86issue
14eissn
0022-538Xissn
1098-5514pii
JVI.00376-14journal_volume
88pub_type
杂志文章abstract:UNLABELLED:Enterovirus 71 (EV71) recruits various cellular factors to assist in the replication and translation of its genome. Identification of the host factors involved in the EV71 life cycle not only will enable a better understanding of the infection mechanism but also has the potential to be of use in the developm...
journal_title:Journal of virology
pub_type: 杂志文章
doi:10.1128/JVI.01677-15
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abstract::Panicum mosaic virus (PMV) is a recently molecularly characterized RNA virus with the unique feature of supporting the replication of two subviral RNAs in a few species of the family Gramineae. The subviral agents include a satellite RNA (satRNA) that is devoid of a coding region and the unrelated satellite panicum mo...
journal_title:Journal of virology
pub_type: 杂志文章
doi:10.1128/JVI.75.11.5429-5432.2001
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abstract::Our previous observations indicated that upon infection with minute virus of mice (MVM), Ehrlich ascites cells lose a transcription elongation activity which is essential for the readthrough of the MVM attenuator. This was monitored by the ability of extracts from uninfected but not from infected cells to support read...
journal_title:Journal of virology
pub_type: 杂志文章
doi:10.1128/JVI.68.4.2741-2745.1994
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abstract::The C-terminal half of the replicase ORF1a polyprotein of the arterivirus equine arteritis virus is processed by a chymotrypsinlike serine protease (SP) (E. J. Snijder et al., J. Biol. Chem. 271:4864-4871, 1996) located in nonstructural protein 4 (nsp4). Three probable SP cleavage sites had previously been identified ...
journal_title:Journal of virology
pub_type: 杂志文章
doi:10.1128/JVI.71.12.9313-9322.1997
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abstract::Using less stringent hybridization conditions and cloned viral DNA probes representing the avian sarcoma virus gag, pol, env, and long terminal repeat (LTR) gene sequences, we detected related sequences in two avian species purportedly lacking all endogenous avian leukosis viruses, the ev- chicken and the Japanese qua...
journal_title:Journal of virology
pub_type: 杂志文章
doi:10.1128/JVI.59.3.669-675.1986
更新日期:1986-09-01 00:00:00
abstract::Hepatitis B viruses (HBVs) replicate by reverse transcription of an RNA intermediate. Packaging of this RNA pregenome into nucleocapsids and replication initiation depend crucially on the interaction of the reverse transcriptase, P protein, with the cis-acting, 5' end-proximal encapsidation signal epsilon. The overall...
journal_title:Journal of virology
pub_type: 杂志文章
doi:10.1128/JVI.71.7.4971-4980.1997
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abstract::Equine infectious anemia virus was found to be comprised of fourteen polypeptides of molecular weight ranging from 10,000 to 79,000. Eighty percent of the virion protein was accounted for by five polypeptides, including two non-glycosylated components (p29 and p13) comprising one-half of the virion protein and three g...
journal_title:Journal of virology
pub_type: 杂志文章
doi:10.1128/JVI.28.3.997-1001.1978
更新日期:1978-12-01 00:00:00
abstract::Integrins mediate cell adhesion and motility on the extracellular matrix, yet they also promote viral attachment and/or entry. Evidence is presented that adenovirus internalization by alpha(v) integrins requires activation of phosphoinositide-3-OH kinase (PI3K), whereas alpha(v) integrin-mediated cell motility depends...
journal_title:Journal of virology
pub_type: 杂志文章
doi:10.1128/JVI.72.3.2055-2061.1998
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journal_title:Journal of virology
pub_type: 杂志文章
doi:10.1128/JVI.70.1.23-29.1996
更新日期:1996-01-01 00:00:00
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journal_title:Journal of virology
pub_type: 杂志文章
doi:10.1128/JVI.61.6.2033-2037.1987
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journal_title:Journal of virology
pub_type: 杂志文章
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abstract::The nonstructural 4B (NS4B) protein of hepatitis C virus (HCV) plays a central role in the formation of the HCV replication complex. To gain insight into the role of charged residues for NS4B function in HCV RNA replication, alanine substitutions were engineered in place of 28 charged residues residing in the N- and C...
journal_title:Journal of virology
pub_type: 杂志文章
doi:10.1128/JVI.00858-11
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abstract::Ocular herpes simplex keratitis (HSK) is a consequence of viral reactivations from trigeminal ganglia (TG) and occurs almost exclusively in the same eye in humans. In our murine oro-ocular (OO) model, herpes simplex virus 1 (HSV-1) inoculation in one side of the lip propagates virus to infect the ipsilateral TG. Repli...
journal_title:Journal of virology
pub_type: 杂志文章
doi:10.1128/JVI.01586-19
更新日期:2019-11-26 00:00:00
abstract::The iridovirus frog virus 3 (FV3) can replicate in culture in fat head minnow (FHM) fish cells or in BHK-21 hamster cells. Viral DNA replication commences about 3 h after infection of FHM cells with FV3. Between 3 and 6 h postinfection (p.i.), a portion of the intranuclear FV3 DNA is partly unmethylated. At later time...
journal_title:Journal of virology
pub_type: 杂志文章
doi:10.1128/JVI.67.12.6973-6978.1993
更新日期:1993-12-01 00:00:00
abstract::A versatile green fluorescent protein (GFP) expression cassette containing the replication origins of the monopartite begomovirus Tomato yellow leaf curl Sardinia virus (TYLCSV) is described. Transgenic Nicotiana benthamiana plants containing one copy of the cassette stably integrated into their genome were superinfec...
journal_title:Journal of virology
pub_type: 杂志文章
doi:10.1128/JVI.80.7.3624-3633.2006
更新日期:2006-04-01 00:00:00
abstract::The antiviral activity of the double-stranded complex poly(rI) . poly(rC) in cell culture was restored or even surpassed if the constituent homopolymers were administered separately. Poly(rI) primed the cells for the antiviral activity of poly(rC) and poly(rC) primed for poly(rI), but neither poly(rI) nor poly(rC) pri...
journal_title:Journal of virology
pub_type: 杂志文章
doi:10.1128/JVI.9.5.721-731.1972
更新日期:1972-05-01 00:00:00
abstract::The 70S RNA of Rous sarcoma virus contains 4S RNAs which serve as primers for the initiation of DNA synthesis in vitro by the RNA-directed DNA polymerase of the virus. We purified these primers in three different ways-by isolation of the covalent complex between primer and nascent DNA, by differential melting of the 7...
journal_title:Journal of virology
pub_type: 杂志文章
doi:10.1128/JVI.13.5.1134-1142.1974
更新日期:1974-05-01 00:00:00
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journal_title:Journal of virology
pub_type: 杂志文章
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journal_title:Journal of virology
pub_type: 杂志文章
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journal_title:Journal of virology
pub_type: 杂志文章
doi:10.1128/JVI.64.5.2033-2040.1990
更新日期:1990-05-01 00:00:00
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journal_title:Journal of virology
pub_type: 杂志文章
doi:10.1128/JVI.67.10.5786-5791.1993
更新日期:1993-10-01 00:00:00
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journal_title:Journal of virology
pub_type: 杂志文章
doi:10.1128/jvi.76.9.4547-4558.2002
更新日期:2002-05-01 00:00:00
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journal_title:Journal of virology
pub_type: 杂志文章
doi:10.1128/jvi.78.6.2921-2934.2004
更新日期:2004-03-01 00:00:00
abstract::Hepatitis A virus (HAV) 3C proteinase expressed in Escherichia coli was purified to homogeneity, and its cleavage specificity towards various parts of the viral polyprotein was analyzed. Intermolecular cleavage of the P2-P3 domain of the HAV polyprotein gave rise to proteins 2A, 2B, 2C, 3ABC, and 3D, suggesting that i...
journal_title:Journal of virology
pub_type: 杂志文章
doi:10.1128/JVI.69.3.1727-1733.1995
更新日期:1995-03-01 00:00:00
abstract::Relatively little is known at the functional genomic level about the global host response to human immunodeficiency virus type 1 (HIV-1) infection. Microarray analyses by several laboratories, including our own, have revealed that HIV-1 infection causes significant changes in host mRNA abundance and regulation of seve...
journal_title:Journal of virology
pub_type: 杂志文章
doi:10.1128/JVI.00288-07
更新日期:2007-07-01 00:00:00
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journal_title:Journal of virology
pub_type: 杂志文章
doi:10.1128/JVI.80.2.671-681.2006
更新日期:2006-01-01 00:00:00
abstract::The cellular tropism of the feline immunodeficiency virus (FIV) is affected by changes in variable region 3 (V3) of the surface (SU) envelope glycoprotein (Verschoor, E. J., et al., J. Virol. 69:4752-4757, 1995). By using high-dose DNA transfection, an FIV molecular clone with a non-CRFK-tropic V3 acquired the ability...
journal_title:Journal of virology
pub_type: 杂志文章
doi:10.1128/JVI.71.9.7132-7135.1997
更新日期:1997-09-01 00:00:00
abstract::Coronaviruses generally have a narrow host range, infecting one or just a few species. Using targeted RNA recombination, we constructed a mutant of the coronavirus mouse hepatitis virus (MHV) in which the ectodomain of the spike glycoprotein (S) was replaced with the highly divergent ectodomain of the S protein of fel...
journal_title:Journal of virology
pub_type: 杂志文章
doi:10.1128/jvi.74.3.1393-1406.2000
更新日期:2000-02-01 00:00:00
abstract::Live attenuated RNA viruses make highly efficient vaccines. Among them, measles virus (MV) vaccine has been given to a very large number of children and has been shown to be highly efficacious and safe. Therefore, this vaccine might be a very promising vector to immunize children against both measles and other infecti...
journal_title:Journal of virology
pub_type: 杂志文章
doi:10.1128/jvi.77.21.11546-11554.2003
更新日期:2003-11-01 00:00:00
abstract::We have studied protein synthesis in cultured cells infected with the six noncytopathic (nc) mutants of the Australia-Victoria strain (AV-WT) of Newcastle disease virus and their plaque-forming revertants. Virus-specific polypeptides accumulated at 30 to 63% of wild-type levels in nc mutant-infected cells and between ...
journal_title:Journal of virology
pub_type: 杂志文章
doi:10.1128/JVI.40.3.691-702.1981
更新日期:1981-12-01 00:00:00