Human immunodeficiency virus Tat transactivation: induction of a tissue-specific enhancer in a nonpermissive cell line

Abstract

The enhancer of the human neurotropic papovavirus JC virus (JCV) restricts viral transcription to glial cells. We utilized the tissue specificity of the JCV enhancer as a tool to investigate the function of human immunodeficiency virus (HIV) Tat in transcriptional activation. The reporter plasmid pJCTAR-CAT was constructed by inserting the HIV type 1 Tat-responsive element, TAR, between the JCV promoter and the chloramphenicol acetyltransferase (CAT) gene. Cotransfection of pJCTAR-CAT and pSV-Tat, an expression vector for Tat, resulted in a 50-fold increase in JCV promoter activity in cells nonpermissive for JCV expression. Both the 98-bp JCV enhancer and the HIV TAR sequences were required for transactivation of pJCTAR-CAT in nonpermissive cells. The transactivation by Tat occurred at the level of transcription, as the increase in CAT activity paralleled an increase in the steady-state levels of CAT mRNA in S1 nuclease and nuclear run-on analyses. In the presence of Tat, the JCV enhancer is functional in cells normally nonpermissive for JCV expression; therefore, our results provide unique evidence that HIV type 1 Tat may regulate the activity of specific transcription factors.