Damagnez V, Mäkelä T P, Cottarel G
Mitotix Inc., Cambridge, MA 02139, USA.
EMBO J. 1995 Dec 15;14(24):6164-72. doi: 10.1002/j.1460-2075.1995.tb00307.x.
The cyclin-dependent kinase (CDK)-activating kinase, CAK, from mammals and amphibians consists of MO15/CDK7 and cyclin H, a complex which has been identified also as a RNA polymerase II C-terminal domain (CTD) kinase. While the Schizosaccharomyces pombe cdc2 gene product also requires an activating phosphorylation, the enzyme responsible has not been identified. We have isolated an essential S.pombe gene, mop1, whose product is closely related to MO15 and to Saccharomyces cerevisiae Kin28. The functional similarity of Mop1 and MO15 is reflected in the ability of MO15 to rescue a mop1 null allele. This suggests that Mop1 would be a CDK, and indeed Mop1 associates with a previously characterized cyclin H-related cyclin Mcs2 of S.pombe. Also, Mop1 and Mcs2 can associate with the heterologous partners human cyclin H and MO15, respectively. Moreover, the rescue of a temperature-sensitive mcs2 strain by expression of mop1+ demonstrates a genetic interaction between mop1 and mcs2. In a functional assay, immunoprecipitated Mop1-Mcs2 acts both as an RNA polymerase II CTD kinase and as a CAK. The CAK activity of Mop1-Mcs2 distinguishes it from the related CDK-cyclin pair Kin28-Ccl1 from S.cerevisiae, and supports the notion that Mop1-Mcs2 may represent a homolog of MO15-cyclin H in S.pombe with apparent dual roles as a RNA polymerase CTD kinase and as a CAK.
哺乳动物和两栖动物的细胞周期蛋白依赖性激酶(CDK)激活激酶CAK由MO15/CDK7和细胞周期蛋白H组成,该复合物也被鉴定为RNA聚合酶II C末端结构域(CTD)激酶。虽然粟酒裂殖酵母cdc2基因产物也需要激活磷酸化,但负责该反应的酶尚未确定。我们分离出了一个粟酒裂殖酵母必需基因mop1,其产物与MO15以及酿酒酵母Kin28密切相关。MO15拯救mop1无效等位基因的能力反映了Mop1和MO15的功能相似性。这表明Mop1可能是一种CDK,实际上Mop1与粟酒裂殖酵母中先前鉴定的与细胞周期蛋白H相关的细胞周期蛋白Mcs2相关联。此外,Mop1和Mcs2可以分别与异源伴侣人细胞周期蛋白H和MO15相关联。此外,通过表达mop1+拯救温度敏感型mcs2菌株证明了mop1和mcs2之间的遗传相互作用。在功能测定中,免疫沉淀的Mop1-Mcs2既作为RNA聚合酶II CTD激酶又作为CAK起作用。Mop1-Mcs2的CAK活性将其与酿酒酵母中相关的CDK-细胞周期蛋白对Kin28-Ccl1区分开来,并支持这样一种观点,即Mop-Mcs2可能代表粟酒裂殖酵母中MO15-细胞周期蛋白H的同源物,具有作为RNA聚合酶CTD激酶和CAK的明显双重作用。