Inhibition of I kappa B kinase activity by sodium salicylate in vitro doesnot reflect its inhibitory mechanism in intact cells

Sodium salicylate inhibits activation of the transcription factor NF-kappa B by blocking the phosphorylation and degradation of the NF-kappa B inhibit or I kappa B alpha. We previously demonstrated that salicylate inhibits I k appa B alpha degradation induced by tumor necrosis factor (TNF) but not by interleukin-1 (IL-1) and implicated p38 mitogen-activated protein kinase ac tivation by salicylate in the inhibition of TNF-induced I kappa B alpha pho sphorylation. Both TNF and IL-1 rapidly activate the I kappa B kinase (IKK) complex, containing the catalytic subunits IKK alpha and IKK beta, which d irectly phosphorylates I kappa B proteins. Others have recently suggested t hat salicylate inhibits NF-kappa B activation by directly binding to IKK be ta. To clarify the mechanism whereby salicylate inhibits IKK activity, we e xamined its effects upon cytokine-induced IKK activity in intact cells and in vitro, Treatment of intact cells with salicylate inhibited TNF-induced b ut not IL-1-induced IKK activity, and this inhibition was prevented by the p38 inhibitor SB203580. In contrast, inhibition of IKK activity by salicyla te in vitro was neither selective for TNF nor affected by SB203580. In vitr o, salicylate treatment comparably inhibited the kinase activity of overexp ressed IKK alpha and IKK beta and also decreased p38 kinase activity. There fore, direct inhibition of IRK activity in vitro does not reflect the inhib itory mechanism of salicylate in intact cells, which involves interference with TNF signaling.