Role of nuclear factor Y in stress-induced activation of the herpes simplex virus type 1 ICP0 promoter.
Herpesviruses are characterized by the ability to establish lifelong latent infections and to reactivate periodically, leading to recurrent disease. The herpes simplex virus type 1 (HSV-1) genome is maintained in a quiescent state in sensory neurons during latency, which is characterized by the absence of detectable viral protein synthesis. Cellular factors induced by stress may act directly on promoters within the latent viral genome to induce the transcription of viral genes and trigger reactivation. In order to identify which viral promoters are induced by stress and elucidate the cellular mechanism responsible for the induction, we generated a panel of HSV-1 promoter-luciferase constructs and measured their response to heat shock. Of the promoters tested, those of ICP0 and ICP22 were the most strongly upregulated after heat shock. Microarray analysis of lytically infected cells supported the upregulation of ICP0 and ICP22 promoters by heat shock. Mutagenic analysis of the ICP0 promoter identified two regions necessary for efficient heat-induced promoter activity, both containing predicted nuclear factor Y (NF-Y) sites, at bases -708 and -75 upstream of the transcriptional start site. While gel shift analysis confirmed NF-Y binding to both sites, only the site at -708 was important for efficient heat-induced activity. Reverse transcription-PCR analysis of selected viral transcripts in the presence of dominant-negative NF-Y confirmed the requirement for NF-Y in the induction of the ICP0 but not the ICP22 promoter by heat shock in lytically infected cells. These findings suggest that the immediate-early ICP0 gene may be among the first genes to be induced during the early events in HSV-1 reactivation, that NF-Y is important for this induction, and that other factors induce the ICP22 promoter.
Kushnir
Anna S
AS
Department of Molecular and Cellular Biology, The University of Arizona, Tucson, AZ 85712-0106, USA.
Davido
David J
DJ
Schaffer
Priscilla A
PA
D000818:Animals; D023081:CCAAT-Binding Factor / Q000502:physiology*; D002460:Cell Line; D002522:Chlorocebus aethiops; D020869:Gene Expression Profiling; D015967:Gene Expression Regulation, Viral; D017781:Genes, Immediate-Early; D005814:Genes, Viral; D018869:Heat-Shock Response / Q000235:genetics*; D018259:Herpesvirus 1, Human / Q000235:genetics*; D006801:Humans; D017874:Immediate-Early Proteins / Q000235:genetics*; D011401:Promoter Regions, Genetic; D044767:Ubiquitin-Protein Ligases / Q000235:genetics*; D014709:Vero Cells; D014775:Virus Activation
D023081:CCAAT-Binding Factor; C057294:ICP22 protein, human herpesvirus 1; D017874:Immediate-Early Proteins; C402244:nuclear factor Y; D044767:Ubiquitin-Protein Ligases; C051804:Vmw110 protein, Human herpesvirus 1