The Varicella-Zoster Virus (VZV) ORF9 Protein Interacts with the IE62 Major VZV Transactivator

Cristian Cilloniz; Wallen Jackson; Charles Grose; Donna Czechowski; John Hay; William T Ruyechan

doi:10.1128/JVI.01274-06

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The Varicella-Zoster Virus (VZV) ORF9 Protein Interacts with the IE62 Major VZV Transactivator

Journal article

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The Varicella-Zoster Virus (VZV) ORF9 Protein Interacts with the IE62 Major VZV Transactivator

Cristian Cilloniz, Wallen Jackson, Charles Grose, Donna Czechowski, John Hay and William T Ruyechan

Journal of virology, Vol.81(2), pp.761-774

01/2007

DOI: 10.1128/JVI.01274-06

PMCID: PMC1797441

PMID: 17079304

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Abstract

The varicella-zoster virus (VZV) ORF9 protein is a member of the herpesvirus UL49 gene family but shares limited identity and similarity with the UL49 prototype, herpes simplex virus type 1 VP22. ORF9 mRNA is the most abundantly expressed message during VZV infection; however, little is known concerning the functions of the ORF9 protein. We have found that the VZV major transactivator IE62 and the ORF9 protein can be coprecipitated from infected cells. Yeast two-hybrid analysis localized the region of the ORF9 protein required for interaction with IE62 to the middle third of the protein encompassing amino acids 117 to 186. Protein pull-down assays with GST-IE62 fusion proteins containing N-terminal IE62 sequences showed that amino acids 1 to 43 of the acidic transcriptional activation domain of IE62 can bind recombinant ORF9 protein. Confocal microscopy of transiently transfected cells showed that in the absence of other viral proteins, the ORF9 protein was localized in the cytoplasm while IE62 was localized in the nucleus. In VZV-infected cells, the ORF9 protein was localized to the cytoplasm whereas IE62 exhibited both nuclear and cytoplasmic localization. Cotransfection of plasmids expressing ORF9, IE62, and the viral ORF66 kinase resulted in significant colocalization of ORF9 and IE62 in the cytoplasm. Coimmunoprecipitation experiments with antitubulin antibodies indicate the presence of ORF9-IE62-tubulin complexes in infected cells. Colocalization of ORF9 and tubulin in transfected cells was visualized by confocal microscopy. These data suggest a model for ORF9 protein function involving complex formation with IE62 and possibly other tegument proteins in the cytoplasm at late times in infection.

Genome and Regulation of Viral Gene Expression

Details

Title: Subtitle: The Varicella-Zoster Virus (VZV) ORF9 Protein Interacts with the IE62 Major VZV Transactivator
Creators: Cristian Cilloniz - Department of Microbiology and Witebsky Center for Microbial Pathogenesis and Immunology, University at Buffalo, Buffalo, New York
Wallen Jackson - Department of Microbiology and Witebsky Center for Microbial Pathogenesis and Immunology, University at Buffalo, Buffalo, New York
Charles Grose - Department of Microbiology and Witebsky Center for Microbial Pathogenesis and Immunology, University at Buffalo, Buffalo, New York
Donna Czechowski - Department of Microbiology and Witebsky Center for Microbial Pathogenesis and Immunology, University at Buffalo, Buffalo, New York
John Hay - Department of Microbiology and Witebsky Center for Microbial Pathogenesis and Immunology, University at Buffalo, Buffalo, New York
William T Ruyechan - Department of Microbiology and Witebsky Center for Microbial Pathogenesis and Immunology, University at Buffalo, Buffalo, New York
Resource Type: Journal article
Publication Details: Journal of virology, Vol.81(2), pp.761-774
DOI: 10.1128/JVI.01274-06
PMID: 17079304
PMCID: PMC1797441
NLM abbreviation: J Virol
ISSN: 0022-538X
eISSN: 1098-5514
Publisher: American Society for Microbiology
Language: English
Date published: 01/2007
Academic Unit: Stead Family Department of Pediatrics; Infectious Disease (Pediatrics)
Record Identifier: 9984093230902771

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