ORF66 protein kinase function is required for T-cell tropism of varicella-zoster virus in vivo.
Several functions have been attributed to the serine/threonine protein kinase encoded by open reading frame 66 (ORF66) of varicella-zoster virus (VZV), including modulation of the apoptosis and interferon pathways, down-regulation of major histocompatibility complex class I cell surface expression, and regulation of IE62 localization. The amino acid sequence of the ORF66 protein contains a recognizable conserved kinase domain. Point mutations were introduced into conserved protein kinase motifs to evaluate their importance to ORF66 protein functions. Two substitution mutants were generated, including a G102A substitution, which blocked autophosphorylation and altered IE62 localization, and an S250P substitution, which had no effect on either autophosphorylation or IE62 localization. Both kinase domain mutants grew to titers equivalent to recombinant parent Oka (pOka) in vitro. pOka66G102A had slightly reduced growth in skin, which was comparable to the reduction observed when ORF66 translation was prevented by stop codon insertions in pOka66S. In contrast, infection of T-cell xenografts with pOka66G102A was associated with a significant decrease in infectious virus production equivalent to the impaired T-cell tropism found with pOka66S infection of T-cell xenografts in vivo. Disrupting kinase activity with the G102A mutation did not alter IE62 cytoplasmic localization in VZV-infected T cells, suggesting that decreased T-cell tropism is due to other ORF66 protein functions. The G102A mutation reduced the antiapoptotic effects of VZV infection of T cells. These experiments indicate that the T-cell tropism of VZV depends upon intact ORF66 protein kinase function.
Schaap-Nutt
Anne
A
Department of Pediatrics, Stanford University School of Medicine, 300 Pasteur Drive, Stanford, CA 94305-5208, USA. anne.s.nutt@gmail.com
Sommer
Marvin
M
Che
Xibing
X
Zerboni
Leigh
L
Arvin
Ann M
AM
D000818:Animals; D002460:Cell Line; D006562:Herpes Zoster / Q000821:virology*; D014645:Herpesvirus 3, Human / Q000472:pathogenicity / Q000502:physiology*; D006801:Humans; D051379:Mice; D016513:Mice, SCID; D016366:Open Reading Frames / Q000502:physiology; D011494:Protein Kinases / Q000502:physiology*; D013601:T-Lymphocytes / Q000821:virology*; D014774:Virulence; D014779:Virus Replication
D011494:Protein Kinases