DYSREGULATION THROUGH THE NF-KAPPA-B ENHANCER AND TATA BOX OF THE HUMAN-IMMUNODEFICIENCY-VIRUS TYPE-1 SUBTYPE-E PROMOTER

The global diversity of human immunodeficiency virus type 1 (HIV-1) ge notypes, termed subtypes A to J, is considerable and growing, However, relatively few studies have provided evidence for an associated pheno typic divergence, Recently, we demonstrated subtype-specific functiona l differences within the long terminal repeat (LTR) region of expandin g subtypes (M. A. Montano, V. A. Novitsky, J. T, Blackard, N. L. Cho, D, A. Katzenstein, and M, Essex, J, Virol, 71:8657-8665, 1997), Notabl y, all HIV-1E isolates were observed to contain a defective upstream N F-kappa B site and a unique TATA-TAR region. In this study, we demonst rate that tumor necrosis factor alpha (TNF-alpha) stimulation of the H IV-1E LTR was also impaired, consistent with a defective upstream NF-k appa B site. Furthermore, repair of the upstream NF-kappa B site withi n HIV-1E partially restored TNF-alpha responsiveness. We also show, in gel shift assays, that oligonucleotides spanning the HIV-1E TATA box displayed a reduced efficiency in the assembly of the TBP-TFIIB-TATA c omplex, relative to an HIV-1B TATA oligonucleotide. In transfection as says, the HIV-1E TATA, when changed to the canonical HIV-1B TATA seque nce (ATAAAA-->ATAATA) unexpectedly reduces both heterologous HIV-1B Ta t and cognate HIV-1E Tat activation of an HIV-1E LTR-driven reporter g ene. However, Tat activation, irrespective of subtype, could be rescue d by introducing a cognate HIV-1B TAR. Collectively, these observation s suggest that the expanding HIV-1E genotype has likely evolved an alt ernative promoter configuration with altered NF-kappa B and TATA regul atory signals in contradistinction with HIV-1B.