Kaldis P, Sutton A, Solomon M J
Yale University School of Medicine, Department of Molecular Biophysics and Biochemistry, New Haven, Connecticut 06520-8024, USA.
Cell. 1996 Aug 23;86(4):553-64. doi: 10.1016/s0092-8674(00)80129-4.
Activation of the cyclin-dependent kinases to promote cell cycle progression requires their association with cyclins as well as phosphorylation of a threonine (residue 161 in human p34cdc2). This phosphorylation is carried out by CAK, the Cdk-activating kinase. We have purified and cloned CAK from S. cerevisiae. Unlike CAKs from other organisms, Cak1p is active as a monomer, has full activity when expressed in E. coli, and is not a component of the basal transcription factor, TFIIH. A temperature-sensitive mutation in CAK1 confers a G2 delay accompanied by low Cdc28p protein kinase activity and shows genetic interactions with altered expression of the gene for the major mitotic cyclin, CLB2. Our data raise the intriguing possibility that p40MO15-cyclin H-MAT1, identified as the predominant CAK in vertebrate cell extracts, may not function as a physiological CAK.
细胞周期蛋白依赖性激酶的激活以促进细胞周期进程需要它们与细胞周期蛋白结合以及苏氨酸(人p34cdc2中的第161位残基)的磷酸化。这种磷酸化由CAK(Cdk激活激酶)完成。我们已从酿酒酵母中纯化并克隆了CAK。与其他生物体的CAK不同,Cak1p作为单体具有活性,在大肠杆菌中表达时具有完全活性,并且不是基础转录因子TFIIH的组成部分。CAK1中的温度敏感突变导致G2期延迟,伴有低Cdc28p蛋白激酶活性,并与主要有丝分裂细胞周期蛋白CLB2基因表达改变存在遗传相互作用。我们的数据提出了一个有趣的可能性,即被鉴定为脊椎动物细胞提取物中主要CAK的p40MO15-细胞周期蛋白H-MAT1可能不具有生理CAK的功能。