The HECT Type Ubiquitin Ligase NEDL2 Is Degraded by Anaphase-promoting Complex/Cyclosome (APC/C)-Cdh1, and Its Tight Regulation Maintains the Metaphase to Anaphase Transition
The HECT Type Ubiquitin Ligase NEDL2 Is Degraded by Anaphase-promoting Complex/Cyclosome (APC/C)-Cdh1, and Its Tight Regulation Maintains the Metaphase to Anaphase Transition
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DOI：
10.1074/jbc.m113.472076
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发表时间：
2013-12-13
影响因子：
4.8
通讯作者：
Zhang, Lingqiang
中科院分区：
文献类型：
--
作者：
Lu, Li;Hu, Shaohua;Zhang, Lingqiang
背景：neddl2是HECT型泛素连接酶NEDD4家族的成员，但其功能在很大程度上仍然未知。结果：在有丝分裂退出期，APC/C-Cdh1降解NEDL2并调控中期到后期的转变。结论：NEDL2似乎动态调节有丝分裂的调节。意义：我们的数据提供了APC/C-Cdh1的新底物，并揭示了HECT型泛素连接酶调节有丝分裂的另一种蛋白。nedd4样泛素连接酶2 （nedd2）是一种HECT型泛素连接酶。在层粘连蛋白错误表达的细胞中，NEDL2增强p73转录活性并降解ATR激酶。与其他HECT型泛素连接酶的重要功能相比，对NEDL2的功能和调控的研究较少。利用一抗免疫沉淀和质谱法，我们确定了一系列可能是nedl2相互作用蛋白的潜在蛋白。候选列表包含许多有丝分裂蛋白，特别是包括后期促进复合物/环体（APC/C）的几个亚基和APC/C的激活剂Cdh1。Cdh1在体内和体外都能与NEDL2相互作用。Cdh1识别一个NEDL2破坏盒(R(740)GSL(743))，并在有丝分裂退出时以APC/ c依赖的方式将其降解。过表达Cdh1会降低NEDL2的蛋白水平，而敲低Cdh1会增加NEDL2的蛋白水平，但对NEDL2 mRNA水平没有影响。NEDL2与有丝分裂纺锤体相关，其蛋白水平在有丝分裂时达到最大值。在有丝分裂过程中，NEDL2的功能是必不可少的，因为NEDL2的缺失延长了中期，而过表达会导致染色体滞后。在结肠癌和宫颈癌中均发现NEDL2蛋白和mRNA的表达升高。我们得出结论，当细胞退出有丝分裂时，NEDL2是APC/C-Cdh1的一种新的底物，并作为中期到后期转变的调节剂。它的过度表达可能有助于肿瘤的发生。
Background: NEDL2 is a member of the HECT type ubiquitin ligase NEDD4 family, but its function remains largely unknown. Results: NEDL2 is degraded by APC/C-Cdh1 during mitotic exit and regulates metaphase to anaphase transition. Conclusion: NEDL2 appears to dynamically modulate regulation of mitosis. Significance: Our data provide a novel substrate of APC/C-Cdh1 and reveal an additional protein by which HECT type ubiquitin ligase can regulate mitosis.NEDD4-like ubiquitin ligase 2 (NEDL2) is a HECT type ubiquitin ligase. NEDL2 enhances p73 transcriptional activity and degrades ATR kinase in lamin misexpressed cells. Compared with the important functions of other HECT type ubiquitin ligase, there is less study concerning the function and regulation of NEDL2. Using primary antibody immunoprecipitation and mass spectrometry, we identify a list of potential proteins that are putative NEDL2-interacting proteins. The candidate list contains many of mitotic proteins, especially including several subunits of anaphase-promoting complex/cyclosome (APC/C) and Cdh1, an activator of APC/C. Cdh1 can interact with NEDL2 in vivo and in vitro. Cdh1 recognizes one of the NEDL2 destruction boxes (R(740)GSL(743)) and targets it for degradation in an APC/C-dependent manner during mitotic exit. Overexpression of Cdh1 reduces the protein level of NEDL2, whereas knockdown of Cdh1 increases the protein level of NEDL2 but has no effect on the NEDL2 mRNA level. NEDL2 associates with mitotic spindles, and its protein level reaches a maximum in mitosis. The function of NEDL2 during mitosis is essential because NEDL2 depletion prolongs metaphase, and overexpression of NEDL2 induces chromosomal lagging. Elevated expression of NEDL2 protein and mRNA are both found in colon cancer and cervix cancer. We conclude that NEDL2 is a novel substrate of APC/C-Cdh1 as cells exit mitosis and functions as a regulator of the metaphase to anaphase transition. Its overexpression may contribute to tumorigenesis.