Characterization of the herpes simplex virion-associated factor responsible for the induction of alpha genes

Abstract

Herpes simplex virus (HSV) genes form three groups, alpha, beta, and gamma, whose synthesis is coordinately regulated and sequentially ordered in a cascade fashion. Earlier studies by Post et al. (Cell 24:555-565, 1981) have shown that chimeric genes constructed by fusion of 5' noncoding leader and upstream sequences of alpha genes to the 5' noncoding leader and structural sequences of the viral thymidine kinase (TK), a beta gene, are regulated as alpha genes upon recombination into the viral genome. In cells converted from TK- to TK+ phenotype, these chimeric genes are induced by infection with homologous TK- virus. The induction of the resident chimeric gene does not require viral protein synthesis and is independent of the presence of functional alpha gene 4 product required for the expression of beta genes. In this paper, we report on the properties of the alpha-TK gene chimera resident in converted TK+ murine (L316) and human (I316) cells. Our results were as follows. (i) The pattern of induction of L316 cells exposed to 0.1, 1.0, and 10 PFU per cell suggested that exposure to competent virus is required for induction and that in untreated preparations this virus corresponds to infectious virus. (ii) UV light-irradiated virus was just as effective as untreated virus in inducing alpha-TK chimeras. (iii) HSV-1(HFEM)tsB7 induced the alpha-TK gene chimeras at the nonpermissive (39 degrees C) temperature; at 39 degrees C the parental HSV-1(HFEM)tsB7 capsids accumulate at nuclear pores and do not release viral DNA. (iv) The alpha-TK gene chimeras were not induced by infection with spontaneous TK- mutants of pseudorabies virus and bovine mammillitis virus or with human cytomegalovirus or adenovirus type 2 or by exposure to lysates of HSV-1-infected cells from which the virus was removed by centrifugation. These results indicate that the alpha gene inducer is a virion component located outside the capsid and that its function might be to stimulate the transcription of alpha genes by recognizing regulatory sites on viral DNAs or host cell products or both.