De Veylder L, Segers G, Glab N, Casteels P, Van Montagu M, Inzé D
Department of Genetics, Flanders Interuniversity Institute for Biotechnology (VIB), Universiteit Gent, Belgium.
FEBS Lett. 1997 Aug 4;412(3):446-52. doi: 10.1016/s0014-5793(97)00822-3.
In Arabidopsis, two cyclin-dependent kinases (CDK), Cdc2aAt and Cdc2bAt, have been described. Here, we have used the yeast two-hybrid system to identify Arabidopsis proteins interacting with Cdc2aAt. Three different clones were isolated, one of which encodes a Suc1/Cks1 homologue. The functionality of the Arabidopsis Suc1/Cks1 homologue, designed Cks1At, was demonstrated by its ability to rescue the temperature-sensitive cdc2-L7 strain of fission yeast at low and intermediate expression levels. In contrast, high cks1At expression levels inhibited cell division in both mutant and wild-type yeast strains. Cks1At binds both Cdc2aAt and Cdc2bAt in vivo and in vitro. Furthermore, we demonstrate that the fission yeast Suc1 binds Cdc2aAt but only weakly Cdc2bAt, whereas the human CksHs1 associated exclusively with Cdc2aAt.
在拟南芥中,已发现两种细胞周期蛋白依赖性激酶(CDK),即Cdc2aAt和Cdc2bAt。在此,我们利用酵母双杂交系统来鉴定与Cdc2aAt相互作用的拟南芥蛋白。分离出了三个不同的克隆,其中一个编码Suc1/Cks1同源物。通过在低表达和中等表达水平下拯救裂殖酵母的温度敏感型cdc2-L7菌株的能力,证明了拟南芥Suc1/Cks1同源物(命名为Cks1At)的功能。相比之下,Cks1At的高表达水平在突变型和野生型酵母菌株中均抑制细胞分裂。Cks1At在体内和体外均能与Cdc2aAt和Cdc2bAt结合。此外,我们证明裂殖酵母Suc1能与Cdc2aAt结合,但与Cdc2bAt的结合较弱,而人类CksHs1仅与Cdc2aAt相关。