Descombes P, Nigg E A
Department of Molecular Biology, University of Geneva, Science II, 30, quai Ernest-Ansermet, CH-1211 Geneva 4, Switzerland.
EMBO J. 1998 Mar 2;17(5):1328-35. doi: 10.1093/emboj/17.5.1328.
Polo-like kinases (Plks), named after the Drosophila gene product polo, have been implicated in the regulation of multiple aspects of mitotic progression, including the activation of the Cdc25 phosphatase, bipolar spindle formation and cytokinesis. Genetic analyses performed in yeast and Drosophila suggest a function for Plks at late stages of mitosis, but biochemical data to support such a function in vertebrate organisms are lacking. Here we have taken advantage of Xenopus egg extracts for exploring the function of Plx1, a Xenopus Plk, during the cell cycle transition from M phase to interphase (I phase). We found that the addition of a catalytically inactive Plx1 mutant to M phase-arrested egg extracts blocked their Ca2+-induced release into interphase. Concomitantly, the proteolytic destruction of several targets of the anaphase-promoting complex and the inactivation of the Cdc2 protein kinase (Cdk1) were prevented. Moreover, the M to I phase transition could be abolished by immunodepletion of Plx1, but was restored upon the addition of recombinant Plx1. These results demonstrate that the exit of egg extracts from M phase arrest requires active Plx1, and they strongly suggest an important role for Plx1 in the activation of the proteolytic machinery that controls the exit from mitosis.
Polo样激酶(Plks),以果蝇基因产物polo命名,参与有丝分裂进程多个方面的调控,包括Cdc25磷酸酶的激活、双极纺锤体形成和胞质分裂。在酵母和果蝇中进行的遗传分析表明Plks在有丝分裂后期发挥作用,但缺乏支持其在脊椎动物中具有这种功能的生化数据。在此,我们利用非洲爪蟾卵提取物来探究非洲爪蟾Plk即Plx1在从M期到间期(I期)的细胞周期转变过程中的功能。我们发现,向处于M期阻滞的卵提取物中添加催化失活的Plx1突变体可阻止其因Ca2+诱导而释放到间期。与此同时,后期促进复合物的几个靶标的蛋白水解破坏以及Cdc2蛋白激酶(Cdk1)的失活也受到了抑制。此外,通过免疫去除Plx1可消除从M期到I期的转变,但添加重组Plx1后可恢复。这些结果表明,卵提取物从M期阻滞中退出需要有活性的Plx1,并且强烈提示Plx1在激活控制有丝分裂退出的蛋白水解机制中起重要作用。