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Volume 271, Number 34, Issue of August 23, 1996 pp. 20820-20827
©1996 by The American Society for Biochemistry and Molecular Biology, Inc.

An NF-kappa B Site in the 5'-Untranslated Leader Region of the Human Immunodeficiency Virus Type 1 Enhances the Viral Expression in Response to NF-kappa B-activating Stimuli

(Received for publication, February 22, 1996, and in revised form, May 31, 1996)

Massimo Mallardo , Emilia Dragonetti , Francesca Baldassarre , Concetta Ambrosino § , Giuseppe Scala § and Ileana Quinto §

From the Dipartimento di Biochimica e Biotecnologie Mediche, Università degli Studi Federico II di Napoli, 80131 Naples and the § Dipartimento di Medicina Sperimentale e Clinica, Università degli Studi di Reggio Calabria, 88100 Catanzaro, Italy

The 5'-untranslated leader region of human immunodeficiency virus, type 1 (HIV-1), includes a complex array of putative regulatory elements whose role in the viral expression is not completely understood. Here we demonstrate the presence of an NF-kappa B-responsive element in the trans-activation response (TAR) region of HIV-1 that confers the full induction of HIV-1 long terminal repeat (LTR) in response to NF-kappa B-activating stimuli, such as DNA alkylating agents, phorbol 12-myristate 13-acetate, and tumor necrosis factor-alpha . The TAR NF-kappa B site GGGAGCTCTC spans from positions +31 to +40 and cooperates with the NF-kappa B enhancer upstream of the TATA box in the NF-kappa B-mediated induction of HIV-1 LTR. The conclusion stems from the following observations: (i) deletion of the two NF-kappa B sites upstream of the TATA box reduces, but does not abolish, the HIV-1 LTR activation by NF-kappa B inducers; (ii) deletion or base pair substitutions of the TAR NF-kappa B site significantly reduce the HIV-1 LTR activation by NF-kappa B inducers; (iii) deletions of both the NF-kappa B sites upstream of the TATA box and the TAR NF-kappa B site abolish the activation of HIV-1 LTR in response to NF-kappa B inducers. Moreover, the p50·p65 NF-kappa B complex binds to the TAR NF-kappa B sequence and trans-activates the TAR NF-kappa B-directed expression. The identification of an additional NF-kappa B site in the HIV-1 LTR points to the relevance of NF-kappa B factors in the HIV-1 life cycle.


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