Anaphase-promoting complex/cyclosome controls HEC1 stability
Anaphase-promoting complex/cyclosome controls HEC1 stability
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DOI：
10.1111/j.1365-2184.2010.00712.x
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发表时间：
2011-02-01
影响因子：
8.5
通讯作者：
Fu, Y. -C.
中科院分区：
文献类型：
--
作者：
Li, L.;Zhou, Y.;Fu, Y. -C.
目的：在有丝分裂过程中，染色体的分离需要一个称为动粒的蛋白质结构来连接染色体DNA和纺锤体微管。对动粒组分进行仔细调节以保证成功的细胞分裂是必不可少的。动粒蛋白的消耗、突变或失调导致有丝分裂停滞和/或细胞死亡。据报道，HEC 1（高表达的癌症）是一个动粒蛋白，耗尽，通过RNA干扰，结果在灾难性的有丝分裂exit.Materials和方法和结果：为了研究如何HEC 1蛋白是控制翻译后，我们分析了后期促进复合物/环体（APC/C）-Cdh 1在降解HEC 1蛋白的作用。在这项研究中，我们表明，HEC 1是一个不稳定的蛋白质，可以通过内源性泛素-蛋白酶体系统在HEK 293 T细胞的目标。RNA干扰和体内泛素化实验结果表明，APC/C-hCdh 1 E3连接酶可将HEC 1泛素化并降解。在进化上保守的D-盒的C-末端的功能作为HEC 1的降解决定子，破坏导致对APC/C-Cdh 1介导的降解的抗性。结论：APC/C-Cdh 1调控HEC 1的稳定性，保证细胞周期的正常进行。
Objective:Chromosome segregation during mitosis requires a physically large proteinaceous structure called the kinetochore to generate attachments between chromosomal DNA and spindle microtubules. It is essential for kinetochore components to be carefully regulated to guarantee successful cell division. Depletion, mutation or dysregulation of kinetochore proteins results in mitotic arrest and/or cell death. HEC1 (high expression in cancer) has been reported to be a kinetochore protein, depletion of which, by RNA interference, results in catastrophic mitotic exit.Materials and methods and results:To investigate how HEC1 protein is controlled post-translation, we analysed the role of anaphase-promoting complex/cyclosome (APC/C)-Cdh1 in degradation of HEC1 protein. In this study, we show that HEC1 is an unstable protein and can be targeted by endogenous ubiquitin-proteasome system in HEK293T cells. Results of RNA interference and in vivo ubiquitination assay indicated that HEC1 could be ubiquitinated and degraded by APC/C-hCdh1 E3 ligase. The evolutionally conserved D-box at the C-terminus functioned as the degron of HEC1, destruction of which resulted in resistance to degradation mediated by APC/C-Cdh1. Overexpression of non-degradable HEC1 (D-box destroyed) induced accumulation of cyclin B protein in vivo and triggered mitotic arrest.Conclusion:APC/C-Cdh1 controls stability of HEC1, ensuring normal cell cycle progression.