Inhibition of IkappaB kinase activity by sodium salicylate in vitro does not reflect its inhibitory mechanism in intact cells.
Sodium salicylate inhibits activation of the transcription factor NF-kappaB by blocking the phosphorylation and degradation of the NF-kappaB inhibitor IkappaBalpha. We previously demonstrated that salicylate inhibits IkappaBalpha degradation induced by tumor necrosis factor (TNF) but not by interleukin-1 (IL-1) and implicated p38 mitogen-activated protein kinase activation by salicylate in the inhibition of TNF-induced IkappaBalpha phosphorylation. Both TNF and IL-1 rapidly activate the IkappaB kinase (IKK) complex, containing the catalytic subunits IKKalpha and IKKbeta, which directly phosphorylates IkappaB proteins. Others have recently suggested that salicylate inhibits NF-kappaB activation by directly binding to IKKbeta. To clarify the mechanism whereby salicylate inhibits IKK activity, we examined its effects upon cytokine-induced IKK activity in intact cells and in vitro. Treatment of intact cells with salicylate inhibited TNF-induced but not IL-1-induced IKK activity, and this inhibition was prevented by the p38 inhibitor SB203580. In contrast, inhibition of IKK activity by salicylate in vitro was neither selective for TNF nor affected by SB203580. In vitro, salicylate treatment comparably inhibited the kinase activity of overexpressed IKKalpha and IKKbeta and also decreased p38 kinase activity. Therefore, direct inhibition of IKK activity in vitro does not reflect the inhibitory mechanism of salicylate in intact cells, which involves interference with TNF signaling.
Alpert
D
D
Department of Microbiology, New York University School of Medicine, New York, New York 10016, USA.
Vilcek
J
J
D000818:Animals; D019556:COS Cells; D017871:Calcium-Calmodulin-Dependent Protein Kinases / Q000037:antagonists & inhibitors; D006801:Humans; D020672:I-kappa B Proteins / Q000037:antagonists & inhibitors*; D007093:Imidazoles / Q000494:pharmacology; D007375:Interleukin-1 / Q000494:pharmacology; D020928:Mitogen-Activated Protein Kinases; D011725:Pyridines / Q000494:pharmacology; D012980:Sodium Salicylate / Q000494:pharmacology*; D014409:Tumor Necrosis Factor-alpha / Q000494:pharmacology; D048051:p38 Mitogen-Activated Protein Kinases
D020672:I-kappa B Proteins; D007093:Imidazoles; D007375:Interleukin-1; D011725:Pyridines; D014409:Tumor Necrosis Factor-alpha; D017871:Calcium-Calmodulin-Dependent Protein Kinases; D020928:Mitogen-Activated Protein Kinases; D048051:p38 Mitogen-Activated Protein Kinases; C093642:SB 203580; D012980:Sodium Salicylate