Valay J G, Simon M, Dubois M F, Bensaude O, Facca C, Faye G
Institut Curie-Biologie, Centre Universitaire, Orsay, France.
J Mol Biol. 1995 Jun 9;249(3):535-44. doi: 10.1006/jmbi.1995.0316.
Kin28p, associated with cyclin Ccl1p, is a putative cyclin-dependent kinase (CDK) of the p34cdc2 family in Saccharomyces cerevisiae. Search for mutations co-lethal (syn mutations) with a kin28 thermosensitive mutation (kin28-ts3) has uncovered genetic interactions between gene KIN28 and genes RAD3, SIN4, STI1 and CDC37. The genetic interaction between KIN28 and the CDC37 cell division cycle gene suggests that a connection exists between the activity of CDK-Kin28p and cell-cycle progression. Both RAD3 and SIN4 gene products are implicated in the RNA polymerase II transcription process. Here we show that RNA polymerase II transcription is drastically reduced in a kin28-ts mutant, at restrictive temperature. This impairment correlates with a markedly decreased phosphorylation of the C-terminal domain (CTD) of the largest subunit of RNA polymerase II (Rpb1p). Thus, the Kin28 gene product is required in vivo for RNA polymerase II phosphorylation and transcriptional activity as recently suggested by experiments using an in vitro reconstituted system.
与细胞周期蛋白Ccl1p相关的Kin28p是酿酒酵母中p34cdc2家族的一种假定的细胞周期蛋白依赖性激酶(CDK)。对与kin28温度敏感突变(kin28-ts3)共致死的突变(合成突变)的搜索揭示了基因KIN28与基因RAD3、SIN4、STI1和CDC37之间的遗传相互作用。KIN28与CDC37细胞分裂周期基因之间的遗传相互作用表明,CDK-Kin28p的活性与细胞周期进程之间存在联系。RAD3和SIN4基因产物都参与RNA聚合酶II的转录过程。在这里我们表明,在限制温度下,kin28-ts突变体中RNA聚合酶II的转录急剧减少。这种损伤与RNA聚合酶II最大亚基(Rpb1p)的C末端结构域(CTD)磷酸化的显著降低相关。因此,正如最近使用体外重组系统的实验所表明的,Kin28基因产物在体内是RNA聚合酶II磷酸化和转录活性所必需的。