A conserved cyclin-binding domain determines functional interplay between anaphase-promoting complex-Cdh1 and Cyclin A-Cdk2 during cell cycle progression
A conserved cyclin-binding domain determines functional interplay between anaphase-promoting complex-Cdh1 and Cyclin A-Cdk2 during cell cycle progression
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DOI：
10.1128/mcb.21.11.3692-3703.2001
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发表时间：
2001-06-01
影响因子：
5.3
通讯作者：
Lukas, J
中科院分区：
文献类型：
--
作者：
Sorensen, CS;Lukas, C;Lukas, J
后期促进复合物（APC）泛素连接酶的周期性活性通过靶向细胞周期调节因子进行破坏来决定多个细胞周期转换的进展。在G，IS过渡，磷酸化依赖性解离的Cdh 1激活亚基抑制APC，允许稳定蛋白质所需的后续细胞周期进程。已经鉴定了启动和维持Cdh 1磷酸化的细胞周期蛋白依赖性激酶（CDK）。然而，涉及哪些细胞周期蛋白-CDK复合物的问题一直是一个争论的问题，细胞周期蛋白-CDK如何与APC亚基相互作用的机制仍然没有解决。在这里，我们证实的证据表明，哺乳动物细胞周期蛋白A-Cdk 2阻止非计划APC在S期的再激活，通过展示其周期性的相互作用与Cdh 1在内源性蛋白质的水平。此外，我们在Cdh 1 WD-40结构域内鉴定了一个保守的细胞周期蛋白结合基序，并表明其破坏在体外和体内消除了Cdh 1-细胞周期蛋白A-Cdk 2相互作用，消除了Cdh 1相关的组蛋白H1激酶活性，并削弱了细胞周期蛋白A-Cdk 2对Cdh 1的磷酸化，细胞周期蛋白结合缺陷型Cdh 1的过表达稳定了APC与Cdh 1的相互作用，并诱导细胞周期停滞在G（1）/S转换期延长。相反，细胞周期蛋白结合缺陷型Cdh 1失去了支持细胞周期蛋白A的APC依赖性蛋白水解的能力，但不支持其他APC底物如细胞周期蛋白B和securin Pds 1的蛋白水解。总的来说，这些数据提供了一个机制的解释细胞周期蛋白A-Cdk 2和APC-Cdh 1之间的相互作用和第一个证据表明，Cdh 1可能通过结合特异性底物激活APC。
Periodic activity of the anaphase-promoting complex (APC) ubiquitin ligase determines progression through multiple cell cycle transitions by targeting cell cycle regulators for destruction. At the G,IS transition, phosphorylation-dependent dissociation of the Cdh1-activating subunit inhibits the APC, allowing stabilization of proteins required for subsequent cell cycle progression. Cyclin-dependent kinases (CDKs) that initiate and maintain Cdh1 phosphorylation have been identified. However, the issue of which cyclin-CDK complexes are involved has been a matter of debate, and the mechanism of how cyclin-CDKs interact with APC subunits remains unresolved. Here we substantiate the evidence that mammalian cyclin A-Cdk2 prevents unscheduled APC reactivation during S phase by demonstrating its periodic interaction with Cdh1 at the level of endogenous proteins. Moreover, we identified a conserved cyclin-binding motif within the Cdhl WD-40 domain and show that its disruption abolished the Cdh1-cyclin A-Cdk2 interaction, eliminated Cdh1-associated histone H1 kinase activity, and impaired Cdhl phosphorylation by cyclin A-Cdk2 in vitro and in vivo, Overexpression of cyclin binding-deficient Cdhl stabilized the APC-Cdh1 interaction and induced prolonged cell cycle arrest at the G(1)/S transition. Conversely, cyclin binding-deficient Cdhl lost its capability to support APC-dependent proteolysis of cyclin A but not that of other APC substrates such as cyclin B and securin Pds1. Collectively, these data provide a mechanistic explanation for the mutual functional interplay between cyclin A-Cdk2 and APC-Cdh1 and the first evidence that Cdhl may activate the APC by binding specific substrates.