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J Virol. 1992 May; 66(5): 2916-2927

Herpes simplex viruses with mutations in the gene encoding ICP0 are defective in gene expression.

J Chen and S Silverstein

Department of Microbiology, Columbia University, New York, New York 10032.

ABSTRACT

Herpes simplex virus type 1 (HSV-1) mutants with codon insertions and deletions in IE-0, the gene encoding ICP0, were constructed. The HSV-1 deletion mutant dl1403 (N. D. Stow and E. C. Stow, J. Gen. Virol. 67:2571-2585, 1986) and an IE-0:lacZ transplacement vector isolated in this study were used to facilitate the construction of mutant viruses. Mutant viruses, all of which produced stable ICP0, were examined for their ability to plaque and grow on both Vero and HeLa cells because previous results showed that HSV-1 immediate-early (IE) gene promoters and their products are differentially expressed in these cells (J. Chen, X. Zhu, and S. Silverstein, Virology 180:207-220, 1991; I. H. Gelman and S. Silverstein, J. Virol. 61:2286-2296, 1987). Viruses with IE-0 genes that only poorly activated reporter genes in transient expression assays plaqued less efficiently on Vero cells and consistently accumulated decreased levels of late proteins. These mutants were also examined in single-step growth curve experiments and for the dependence of virus yield on multiplicity of infection (MOI). At low MOIs, their yields were less in Vero cells than in HeLa cells; by contrast, at high MOIs, there was no apparent difference in yield in either cell type, although each virus produced considerably fewer progeny than wild-type virus. Analysis of steady-state levels of RNA from genes representing each of the three major kinetic classes demonstrated that lower levels of RNAs accumulate in these mutants. We conclude from these studies that while ICP0 is not essential for virus growth in tissue culture, defects in this gene result in impairment of virus replication and delay the expression of early and late gene transcripts.


J Virol. 1992 May; 66(5): 2916-2927




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