Dishevelled is a NEK2 kinase substrate controlling dynamics of centrosomal linker proteins
Dishevelled is a NEK2 kinase substrate controlling dynamics of centrosomal linker proteins
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DOI：
10.1073/pnas.1608783113
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发表时间：
2016-08-16
影响因子：
11.1
通讯作者：
Bryja, Vitezslav
中科院分区：
文献类型：
--
作者：
Cervenka, Igor;Valnohova, Jana;Bryja, Vitezslav
Dishevelled（DVL）是Wnt信号通路中的一个关键支架蛋白和分支点。在这里，我们提出了确凿的证据，DVL调节中心体周期。我们证明，DVL凌乱和轴（DIX）结构域，但不是DIX结构域介导的多聚化，是必不可少的DVL的中心体定位。DVL在细胞周期中积累，并与NIMA相关激酶2（NEK 2）结合，NEK 2能够磷酸化DVL的多个残基，如一组新的磷酸化特异性抗体所检测到的。这产生了与CDK 5调节亚基相关蛋白2（CDK 5 RAP 2）和中心体Nek 2相关蛋白1（C-NAP 1）有效结合的界面，这两种蛋白质是中心体连接子。DVL从中心体的置换和其在NEK 2磷酸化时释放到细胞质中与接头蛋白的去除偶联，这是中心体分离和有丝分裂纺锤体的适当形成所必需的事件。DVL的缺乏阻止了NEK 2控制的松散中心体连接体的溶解和随后的中心体分离。增加DVL水平，相反，隔离中心体NEK 2和模仿单极纺锤体缺陷诱导的显性负性版本的激酶。因此，我们的研究揭示了中心体和Wnt信号之间的分子串扰。
Dishevelled (DVL) is a key scaffolding protein and a branching point in Wnt signaling pathways. Here, we present conclusive evidence that DVL regulates the centrosomal cycle. We demonstrate that DVL dishevelled and axin (DIX) domain, but not DIX domain-mediated multimerization, is essential for DVL's centrosomal localization. DVL accumulates during the cell cycle and associates with NIMA-related kinase 2 (NEK2), which is able to phosphorylate DVL at amultitude of residues, as detected by a set of novel phospho-specific antibodies. This creates interfaces for efficient binding to CDK5 regulatory subunit-associated protein 2 (CDK5RAP2) and centrosomal Nek2-associated protein 1 (C-NAP1), two proteins of the centrosomal linker. Displacement of DVL from the centrosome and its release into the cytoplasm on NEK2 phosphorylation is coupled to the removal of linker proteins, an event necessary for centrosomal separation and proper formation of the mitotic spindle. Lack of DVL prevents NEK2-controlled dissolution of loose centrosomal linker and subsequent centrosomal separation. Increased DVL levels, in contrast, sequester centrosomal NEK2 and mimic monopolar spindle defects induced by a dominant negative version of this kinase. Our study thus uncovers molecular crosstalk between centrosome and Wnt signaling.