Journal article
The Requirement of Varicella Zoster Virus Glycoprotein E (gE) for Viral Replication and Effects of Glycoprotein I on gE in Melanoma Cells
Virology (New York, N.Y.), Vol.304(2), pp.176-186
12/20/2002
DOI: 10.1006/viro.2002.1556
PMID: 12504560
Abstract
The glycoprotein E (gE) of varicella zoster virus (VZV), encoded by ORF68, is the most abundant viral glycoprotein. In the current experiments, we demonstrated that ORF68 deletion was incompatible with recovery of infectious virus from VZV cosmids. Replacing ORF68 at a nonnative AvrII site in the genome restored infectivity. Further, we found that VZV gE could be expressed under the control of the Tet-On promoter in stably transfected melanoma cell lines (Met-gE cells) without evidence of toxicity. In these Met-gE cells, gE colocalized with γ-adaptin, a trans Golgi network marker, in perinuclear sites, but did not reach plasma membranes. In order to investigate how infection altered gE localization, we made a recombinant virus, vOka-MSPgE, with ORF68 from the VZV MSP strain. VZV MSP encodes a mutant gE protein (D150N) that lacks the mAb epitope, 3B3 (Santos et al., Virology 275, 306–317, 2000), whereas Met-gE protein binds mAb 3B3. Within 48 h after Met-gE cells were infected with vOka-MSPgE, the steady-state distribution of Met-gE protein extended beyond the perinuclear areas to other cytoplasmic sites and to plasma membranes. A second recombinant, vOka-MSPgE without gI (vOka-MSPgEΔgI), was constructed to investigate Met-gE protein distribution in the absence of gI. The redistribution of Met-gE protein which was observed by 48 h after vOka-MSPgE infection did not occur until 5 days (140 h) within vOka-MSPgEΔgI infected cells. After vOka-MSPgE infection of Met-gE cells, most Met-gE protein was in the final 94K mature form by 72 h. However, progression to predominance of mature gE was delayed in Met-gE cells infected with vOka-MSPgEΔgI. These observations confirm our hypothesis that VZV gE is essential, based upon the demonstration of restored infectivity after replacing ORF68 in a nonnative site in the genome, and provide further evidence of the role of gI in facilitating the maturation and intracellular distribution of this critical VZV glycoprotein.
Details
- Title: Subtitle
- The Requirement of Varicella Zoster Virus Glycoprotein E (gE) for Viral Replication and Effects of Glycoprotein I on gE in Melanoma Cells
- Creators
- Chengjun Mo - Department of Pediatrics, Stanford University School of Medicine, Stanford, California, 94305Jay Lee - Department of Pediatrics, Stanford University School of Medicine, Stanford, California, 94305Marvin Sommer - Department of Pediatrics, Stanford University School of Medicine, Stanford, California, 94305Charles Grose - Department of Pediatrics and Microbiology, University of Iowa College of Medicine, Iowa City, Iowa, 52242Ann M Arvin - Department of Pediatrics, Stanford University School of Medicine, Stanford, California, 94305
- Resource Type
- Journal article
- Publication Details
- Virology (New York, N.Y.), Vol.304(2), pp.176-186
- DOI
- 10.1006/viro.2002.1556
- PMID
- 12504560
- NLM abbreviation
- Virology
- ISSN
- 0042-6822
- eISSN
- 1096-0341
- Publisher
- Elsevier Inc
- Language
- English
- Date published
- 12/20/2002
- Academic Unit
- Stead Family Department of Pediatrics; Infectious Disease (Pediatrics)
- Record Identifier
- 9984093455902771
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