Varicella-zoster virus open reading frame 10 is a virulence determinant in skin cells but not in T cells in vivo.
The open reading frame 10 (ORF10) of varicella-zoster virus (VZV) encodes a tegument protein that enhances transactivation of VZV genes and has homology to herpes simplex virus type 1 (HSV-1) VP16. While VP16 is essential for HSV replication, ORF10 is dispensable for vaccine OKA (VOKA) growth in vitro. We used parent OKA (POKA) cosmids to delete ORF10, producing POKA delta10; point mutations that disrupted the acidic activation domain and the putative motif for binding human cellular factor 1 (HCF-1) in ORF10 protein yielded POKA10-Phe28Ala, POKA10-Phe28Ser, and POKA10-mHCF viruses. Deleting ORF10 or mutating these two functional domains had no effect on VZV replication, immediate-early gene transcription, or virion assembly in vitro. However, deleting ORF10 reduced viral titers and the extent of cutaneous lesions significantly in SCIDhu skin xenografts in vivo compared to POKA. Epidermal cells infected with POKA delta10 had significantly fewer DNA-containing nucleocapsids and complete virions compared to POKA; extensive aggregates of intracytoplasmic viral particles were also observed. Altering the activation or the putative HCF-1 domains of ORF10 protein had no consequences for VZV replication in vivo. Thus, the decreased pathogenic potential of POKA delta10 in skin could not be attributed to absence of these ORF10 protein functions. In contrast to skin cells, deleting ORF10 did not impair VZV T-cell tropism in vivo, as assessed by infectious virus yields. We conclude that ORF10 protein is required for efficient VZV virion assembly and is a specific determinant of VZV virulence in epidermal and dermal cells in vivo.
Department of Pediatrics, Stanford University School of Medicine, Stanford, CA 94305-5208, USA. xibing@stanford.edu
Xibing
X
Che
Leigh
L
Zerboni
Marvin H
MH
Sommer
Ann M
AM
Arvin
D000818:Animals; D003360:Cosmids; D014645:Herpesvirus 3, Human / Q000235:genetics* / Q000472:pathogenicity*; D006801:Humans; D007150:Immunohistochemistry; D008297:Male; D008545:Melanoma / Q000473:pathology / Q000648:ultrastructure / Q000821:virology; D051379:Mice; D016513:Mice, SCID; D016366:Open Reading Frames; D017354:Point Mutation; D011995:Recombination, Genetic; D012867:Skin / Q000473:pathology / Q000648:ultrastructure / Q000821:virology*; D013601:T-Lymphocytes / Q000821:virology*; D013997:Time Factors; D014183:Transplantation, Heterologous; D014613:Vaccines, Attenuated / Q000276:immunology; D014771:Virion / Q000648:ultrastructure; D014774:Virulence; D014779:Virus Replication
D014613:Vaccines, Attenuated