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Journal of Virology, October 1998, p. 8446-8452, Vol. 72, No. 10
0022-538X/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.

Dysregulation through the NF-kappa B Enhancer and TATA Box of the Human Immunodeficiency Virus Type 1 Subtype E Promoter

Monty A. Montano, Christian P. Nixon, and Max Essex*

Department of Immunology and Infectious Diseases, Harvard School of Public Health, Boston, Massachusetts 02115

Received 27 March 1998/Accepted 14 July 1998

The global diversity of human immunodeficiency virus type 1 (HIV-1) genotypes, termed subtypes A to J, is considerable and growing. However, relatively few studies have provided evidence for an associated phenotypic divergence. Recently, we demonstrated subtype-specific functional differences within the long terminal repeat (LTR) region of expanding 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). Notably, all HIV-1E isolates were observed to contain a defective upstream NF-kappa B site and a unique TATA-TAR region. In this study, we demonstrate that tumor necrosis factor alpha (TNF-alpha ) stimulation of the HIV-1E LTR was also impaired, consistent with a defective upstream NF-kappa B site. Furthermore, repair of the upstream NF-kappa B site within 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 complex, relative to an HIV-1B TATA oligonucleotide. In transfection assays, the HIV-1E TATA, when changed to the canonical HIV-1B TATA sequence (ATAAAAright-arrowATATAA) unexpectedly reduces both heterologous HIV-1B Tat and cognate HIV-1E Tat activation of an HIV-1E LTR-driven reporter gene. However, Tat activation, irrespective of subtype, could be rescued by introducing a cognate HIV-1B TAR. Collectively, these observations suggest that the expanding HIV-1E genotype has likely evolved an alternative promoter configuration with altered NF-kappa B and TATA regulatory signals in contradistinction with HIV-1B.

* Corresponding author. Mailing address: Department of Immunology and Infectious Diseases, Harvard School of Public Health, FXB 402, 651 Huntington Ave., Boston, MA 02115. Phone: (617) 432-2334. Fax: (617) 739-8348. E-mail: messex{at}sph.harvard.edu.

Journal of Virology, October 1998, p. 8446-8452, Vol. 72, No. 10
0022-538X/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.


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