The cyclin-dependent kinase (CDK)-activating kinase (CAK) phosphorylates a conserved threonine residue on CDKs and activates them. Two known classes of CAKs are represented by monomeric Cak1p in budding yeast Saccharomyces cerevisiae and by heterotrimeric CDK7-cyclin H-Mat1 in human and other metazoa. We report here the identification of p42, a novel CAK activity in human cells. p42 has sequence homology to both Cak1p and CDK7 groups of CAKs. p42 is essential for the phosphorylation of Thr-160 and activation of CDK2. A dominant-negative p42 mutant, T161A, and posttranscriptional gene silencing of p42 with RNAi-impaired Thr-160 phosphorylation and activity of CDK2. Purified p42 phosphorylated glutathione S-transferase-CDK2 at Thr-160 within the T-loop and activated its histone H1 kinase activity. Finally, p42 is indispensable for cell growth. Cells lacking p42 were incapable of growing and forming colonies whereas cells with a reduced level of p42 grew at significantly slower rates than control cells. Our findings suggest that p42 represents a novel CAK activity in mammalian cells.
The costs of publication of this article were defrayed in part by the payment of page charges. This article must therefore be hereby marked “advertisement” in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.
