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Papers In Press, published online ahead of print February 4, 2002
Webb-Waring Institute for Cancer, Aging and Antioxidant Research, University of Colorado Health Sciences Center, Denver, CO 80262
Corresponding Author: sonia.flores{at}uchsc.edu
Human immunodeficiency virus infection is associated with inflammation and endothelial cell activation that cannot be ascribed to direct infection by the virus or to the presence of opportunistic infections. Factors related to the virus itself, to the host and/or to environmental exposures probably account for these observations. The HIV protein Tat, a viral regulator required for efficient transcription of the viral genome in host cells is secreted from infected cells and taken up by uninfected by-stander cells. Tat can also act as a general transcriptional activator of key inflammatory molecules. We have examined whether Tat contributes to this endothelial cell activation by activating NF-
J. Biol. Chem, 10.1074/jbc.M108591200
Submitted on September 6, 2001
Revised on January 30, 2002
Accepted on February 1, 2002
The human immunodeficiency virus-1 Tat protein activates endothelial cell E-selectin expression via an NF-
-dependent mechanism
B. Human endothelial cells exposed to Tat in the culture medium activated E-selectin expression with delayed kinetics compared to TNF-
. Tat-mediated E-selectin up-regulation required the basic domain of Tat and was inhibited by a Tat antibody. Transfection of E-selectin promoter-reporter constructs into Tat-bearing cells or into endothelial cells co-transfected with a Tat-expression vector resulted in induction of luciferase expression. Either Tat or TNF activated p65 translocation and binding of p65/p50 heterodimers to an oligonucleotide containing the E-selectin B site 3 sequence. Tat-mediated p65 translocation was also delayed compared with TNF. Neither agent induced new synthesis of p65. A super-repressor adenovirus (Ad-
IB S32A/S36A) that constitutively sequesters IB in the cytoplasm as well as cycloheximide or actinomycin D inhibited Tat- or TNF-mediated B translocation and E-selectin up-regulation, suggesting that new macromolecule synthesis is required. These results indicate that Tat, by activating NF-B, increases E-selectin expression and resets the threshold for inflammation.
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