Regulation of the vhs gene promoter of pseudorabies virus by IE180 and EP0, and the requirement of a Sp1 Site for the promoter function

Abstract

The virion host shutoff (vhs) protein is a virion component of Alphaherpesviruses, including pseudorabies virus. In this work, the upstream sequences of vhs gene of pseudorabies virus (TNL strain) was cloned and sequenced. We linked the upstream sequences of vhs gene to the CAT reporter gene and examined the promoter function of this region. The immediate-early protein IE180 of Pseudorabies Virus (PRV) is expressed immediately after infection and plays a vital role in the regulation of other viral genes. Our results demonstrated that the vhs promoter was regulated by the IE180 in a dosage-dependent manner; the vhs promoter was stimulated by low concentration of IE180 but suppressed by high concentration of IE180. Mutational analysis indicated that the only IE180 binding site at the vhs promoter was not essential for its function; however, a Sp1 binding site (15 bp downstream to TATA box) was critical to its function. In addition, the result of cotransfection demonstrated that early protein 0 (EP0) of PRV, another protein with transcriptional function, inhibited the activity of the vhs promoter.