Analysis of the promoter and cis-acting elements regulating expression of herpes simplex virus type 2 latency-associated transcripts

K Wang, P R Krause, S E Straus
1995 Journal of Virology  
In latently infected neurons, herpes simplex virus type 2 (HSV-2) expresses one abundant family of transcripts, the latency-associated transcripts (LATs). We demonstrate here that the sequence lying about 700 bp upstream of the 5 end of the HSV-2 major LAT acts as a very strong promoter in transient expression assays in both neuronal and nonneuronal cells. Transcription starts about 27 to 32 bp downstream of a functional TATA box. The proximal fragment from ؊102 to ؉34 includes the basal
more » ... es the basal promoter and accounts for constitutive transcriptional activity in various cell lines. The distal region from ؊392 to ؊103 contributes to particularly strong promoter activity in neuronal cell lines and involves multiple cis-acting elements. A functional activating transcription factor/cyclic AMP (cAMP) response element binding protein motif lies just upstream of the TATA. By DNase I footprint and methylation protection assays, we identified several additional proteinbinding sites upstream of the activating transcription factor/cAMP response element binding protein motif. A GC-rich element, termed LAT-3, was located between bases ؊128 to ؊102. A 2-bp substitution in LAT-3 markedly reduced promoter activity and abolished protein-binding ability in vitro. Gel retardation assay showed no competition for protein binding to LAT-3 by other GC-rich elements. LAT-3 appears to be a novel cis-acting element that may contribute to the neuronal responsiveness of the HSV-2 LAT promoter.
doi:10.1128/jvi.69.5.2873-2880.1995 fatcat:ahg63mbkpffanfrkdaahfn7t6u