Carr A M, MacNeill S A, Hayles J, Nurse P
Department of Biochemistry, University of Oxford, UK.
Mol Gen Genet. 1989 Jul;218(1):41-9. doi: 10.1007/BF00330563.
The cdc2+ gene function plays a central role in the control of the mitotic cell cycle of the fission yeast Schizosaccharomyces pombe. Recessive temperature-sensitive mutations in the cdc2 gene cause cell cycle arrest when shifted to the restrictive temperature, while a second class of mutations within the cdc2 gene causes a premature advancement into mitosis. Previously the cdc2+ gene has been cloned and has been shown to encode a 34 kDa phosphoprotein with in vitro protein kinase activity. Here we describe the cloning of 11 mutant alleles of the cdc2 gene using two simple methods, one of which is presented here for the first time. We have sequenced these alleles and find a variety of single amino acid substitutions mapping throughout the cdc2 protein. Analysis of these mutations has identified a number of regions within the cdc2 protein that are important for cdc2+ activity and regulation. These include regions which may be involved in the interaction of the cdc2+ gene product with the proteins encoded by the wee1+, cdc13+ and suc1+ genes.
cdc2+基因功能在粟酒裂殖酵母有丝分裂细胞周期的调控中起着核心作用。cdc2基因中的隐性温度敏感突变在转移到限制温度时会导致细胞周期停滞，而cdc2基因内的第二类突变会导致过早进入有丝分裂。此前，cdc2+基因已被克隆，并已证明其编码一种具有体外蛋白激酶活性的34 kDa磷蛋白。在这里，我们描述了使用两种简单方法克隆cdc2基因的11个突变等位基因，其中一种方法是首次在此介绍。我们对这些等位基因进行了测序，发现了分布在整个cdc2蛋白上的各种单氨基酸取代。对这些突变的分析确定了cdc2蛋白内一些对cdc2+活性和调控很重要的区域。这些区域包括可能参与cdc2+基因产物与wee1+、cdc13+和suc1+基因编码的蛋白质相互作用的区域。
