Affinity purification reveals the association of WD40 protein constitutive photomorphogenic 1 with the hetero-oligomeric TCP-1 chaperonin complex in mammalian cells.
Affinity purification reveals the association of WD40 protein constitutive photomorphogenic 1 with the hetero-oligomeric TCP-1 chaperonin complex in mammalian cells.
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亲和纯化揭示了哺乳动物细胞中 WD40 蛋白组成型光形态发生 1 与异源寡聚 TCP-1 伴侣蛋白复合物的关联。
DOI：
10.1016/j.biocel.2005.12.019
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发表时间：
2006
期刊：
影响因子：
--
通讯作者：
Deng,XingWang
中科院分区：
文献类型：
--
作者：
Yi,Chunling;Li,Shuting;Wang,Jian;Wei,Ning;Deng,XingWang
构成光形态发生蛋白1(COP1)是一种由无名指、卷曲结构域和7个WD40重复序列组成的蛋白质，其功能类似于E3泛素连接酶，靶向于高等植物和哺乳动物细胞中泛素化和降解的关键转录因子。虽然COP1在植物的光介导发育中是必需的，但它在哺乳动物中的对应物已经与肿瘤的发生有关。我们之前已经证明，COP1在哺乳动物细胞中形成高分子量的复合体。在这里，我们报告了我们试图通过亲和纯化和质谱分析相结合的方法来表征哺乳动物COP1复合体的组成。我们发现，瞬时和稳定表达的COP1都与异寡聚的TCP-1伴侣蛋白复合体(TIC)、热休克蛋白70(Hsp70)和BAG家族分子伴侣调节因子-2(Bag2)有关。此外，稳定表达的COP1还与主要的vault蛋白(MVP)和易位的启动子区域(Tpr)结合。已知TIC/Hsp70复合体与酿酒酵母中的许多WD40蛋白相互作用并协助其折叠。在哺乳动物细胞中，WD40蛋白COP1与TIC/Hsp70的结合表明，促进WD40蛋白的折叠可能是TRIC/Hsp70从酵母到哺乳动物的一种保守功能。
Constitutive photomorphogenic 1 (COP1), a protein composed of a RING finger, a coiled-coil domain and seven WD40 repeats, functions as an E3 ubiquitin ligase that targets key transcription factors for ubiquitination and degradation in both higher plants and mammalian cells. While COP1 is required for light-mediated development in plants, its mammalian counterpart has been implicated in tumorigenesis. We previously showed that COP1 forms high-molecular-weight complexes in mammalian cells. Here we report our attempts in characterizing the components of the mammalian COP1 complexes by affinity purification combined with mass spectral analysis. We find that both transiently and stably expressed COP1 associates with the hetero-oligomeric TCP-1 chaperonin complex (TRiC), heat shock protein 70 (Hsp70) and BAG-family molecular chaperone regulator-2 (BAG2). In addition, stably expressed COP1 binds to major vault protein (MVP) and translocated promoter region (Tpr). The TRiC/Hsp70 complex is known to interact with and assist in the folding of a number of WD40 proteins in Saccharomyces cerevisiae. The association of WD40 protein COP1 with TRiC/Hsp70 in mammalian cells suggests that facilitating the folding of WD40 proteins may be a conserved function for TRiC/Hsp70 from yeast to mammals.
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