IKK-GAMMA IS AN ESSENTIAL REGULATORY SUBUNIT OF THE I-KAPPA-B KINASE COMPLEX

Pro-inflammatory cytokines activate the transcription factor NF-kappa B by stimulating the activity of a protein kinase that phosphorylates I kappa B, an inhibitor of NF-kappa B1-5, af sites that trigger its ub iquitination and degradation. This results in the nuclear translocatio n of freed NF-kappa B dimers and the activation of transcription of ta rget genes(6,7). Many of these target genes code for immunoregulatory proteins(8,9). A large, cytokine-responsive I kappa B kinase (IKK) com plex has been purified and the genes encoding two of its subunits have been cloned(1,2,5). These subunits, IKK-alpha and IKK-beta, are prote in kinases whose function is needed for NF-kappa B activation by pro-i nflammatory stimuli. Here, by using a monoclonal antibody against IKK- alpha, we purify the IKK complex to homogeneity from human cell lines. We find that IKK is composed of similar amounts of IKK-alpha, IKK-bet a and two other polypeptides, for which we obtained partial sequences. These polypeptides are differentially processed forms of a third subu nit, IKK-gamma. Molecular cloning and sequencing indicate that IKK-gam ma is composed of several potential coiled-coil moths. IKK-gamma inter acts preferentially with IKK-beta and is required for the activation o f the IKK complex. An IKK-gamma carboxy-terminal truncation mutant tha t still binds IKK-beta blocks the activation of IKK and NF-kappa B.