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Papers In Press, published online ahead of print May 22, 2002
Internal Medicine, University of Texas Southwestern, Dallas, TX 75216
Corresponding Author: lance.terada{at}med.va.gov
We previously found that the angiogenic factors TNF alpha and HIV1 Tat activate an NAD(P)H oxidase in endothelial cells which operates upstream of c-Jun amino terminal kinase (JNK), a MAPK involved in the determination of cell fate. To further understand oxidant-related signaling pathways, we screened lung and endothelial cell libraries for interaction partners of p47phox and recovered the orphan adapter TNF-receptor associated factor 4 (TRAF4). Domain analysis suggested a tail-to-tail interaction between the C-terminus of p47phox and the conserved TRAF domain of TRAF4. In addition, TRAF4, like p47phox, was recovered largely in the cytoskeleton/membrane fraction. Coexpression of p47phox and TRAF4 increased oxidant production and JNK activation, whereas each alone had minimal effect. In addition, a fusion between p47phox and the TRAF4 C-terminus constitutively activated JNK, and this activation was decreased by the antioxidant N-acetyl cysteine. In contrast, overexpression of the p47phox- binding domain of TRAF4 blocked endothelial cell JNK activation by TNF alpha and HIV1 Tat, suggesting an uncoupling of p47phox from upstream signaling events. A secondary screen of endothelial cell proteins for TRAF4-interacting partners yielded a number of proteins known to control cell fate. We conclude that endothelial cell agonists such as TNF alpha and HIV1 Tat initiate signals which enter basic signaling cassettes at the level of TRAF4 and an NAD(P)H oxidase. We speculate that endothelial cells may target endogenous oxidant production to specific sites critical to cytokine signaling as a mechanism for increasing signal specificity and decreasing toxicity of these reactive species.
J. Biol. Chem, 10.1074/jbc.M202665200
Submitted on March 19, 2002
Revised on April 29, 2002
Accepted on May 22, 2002
Involvement of TRAF4 in oxidative activation of c-Jun amino terminal kinase
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