The control of herpes simplex virus type-1 late gene transcription: a 'TATA-box'/cap site region is sufficient for fully efficient regulated activity

Abstract

The transcriptional programme of herpes simplex virus type 1 (HSV-1) is organised into three principle phases; immediate-early (IE), early (E) and late. The appearance of IE gene products provides the switch for E transcription. Abundant expression of late genes requires viral DNA replication. There is some overlap between E and late genes according to their degree of dependence on DNA replication. The pattern of expression of gene US11 is regulated with 'true-late' kinetics (Johnson et al., 1986). In a transient assay system, regulation of a plasmid-borne US11 promoter mimics its viral counterpart, and has a similar dependence on DNA replication for abundant expression. Using plasmids which contain a functional HSV-1 origin of replication (ORIS), we have identified the sequence requirements for the expression of late genes. All DNA sequence elements necessary for fully efficient regulated expression of US11 lie within 31 bp of the RNA cap sites; therefore it appears that a late gene promoter consists only of a proximal 'TATA-box' and cap-site region. We tested this hypothesis by removing the distal upstream region of the gD promoter (which is required for its normal regulation as an early promoter) and linking this truncated promoter to ORIS. This resulted in the conversion of gD promoter regulation to late gene kinetics during virus superinfection. The implications of these results for the mechanisms of HSV gene regulation are discussed.