Rad regulates interleukin 1-induced nuclear factor κB activation in an inhibitory protein κBα-independent manner by enhancing the ability of the p65 subunit to transactivate gene expression

We have examined the involvement of Rac1 in nuclear factor kappa B (NF kappa B) activation by interleukin 1 (IL1). IL1 induced a rapid and sustained activation of Rad in the thymoma cell line EL4.NOB-1. Transient transfection with dominant negative RacN17 inhibited IL1-induced kappa B-dependent reporter gene expression but not I kappa B alpha degradation, whereas constitutively active RacV12 po tentiated KB-dependent reporter gene expression in response to IL1 but had no effects on its own. Using porcine aortic endothelial cells stably transfected with RacV12 or RacN17 under the control of an inducible promoter, we confirmed that RacV12 did not affect I kappa B alpha degradation, nor did RacN17 inhibit the nl-induced response. RacV12 was also unable to induce nuclear translocation of NF kappa B. These effects suggested a role for Rad in p65-mediated transactivation of NF kappa B, independent of I kappa B alpha regulation. In support of this we found that IL1 activated a pathway leading to increased p65 transactivation activity and that RacV12 alone could drive this response in both cell systems. Additionally, RacN17 inhibited IL1-driven p65-mediated transactivation. From data using specific inhibitors of p38 and p42/ p44 kinases we propose that both p38 and p42/p44 lie downstream of Rad on the nl pathway leading to enhanced transactivation by p65.