The Molecular Chaperone TRiC/CCT Binds to the Trp-Asp 40 (WD40) Repeat Protein WDR68 and Promotes Its Folding, Protein Kinase DYRK1A Binding, and Nuclear Accumulation*
The Molecular Chaperone TRiC/CCT Binds to the Trp-Asp 40 (WD40) Repeat Protein WDR68 and Promotes Its Folding, Protein Kinase DYRK1A Binding, and Nuclear Accumulation*
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分子伴侣 TRiC/CCT 与 Trp-Asp 40 (WD40) 重复蛋白 WDR68 结合并促进其折叠、蛋白激酶 DYRK1A 结合和核积累*
DOI：
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发表时间：
2014
影响因子：
4.8
通讯作者：
E. Nishida
中科院分区：
文献类型：
--
作者：
Y. Miyata;Takeshi Shibata;M. Aoshima;T. Tsubata;E. Nishida
背景资料：Trp-Asp（WD）重复蛋白68（WDR 68）是双特异性酪氨酸磷酸化调节蛋白激酶（DYRK 1A）的结合伴侣，DYRK 1A是一种部分负责唐氏综合征的激酶。结果如下：分子伴侣T复合蛋白1（TCP 1）环复合物/含伴侣蛋白的TCP 1（TRiC/CCT）与WDR 68结合，并调节其结构、DYRK 1A结合和细胞定位。结论：TRiC/CCT对WDR 68的正确折叠和功能至关重要。意义：TRiC/CCT可能在形成功能性WD 40重复七叶β螺旋桨结构中具有一般作用。Trp-Asp（WD）repeat protein 68（WDR 68）是一种进化上保守的WD 40重复序列蛋白，其结合多种蛋白，包括双特异性酪氨酸磷酸化调节蛋白激酶（DYRK 1A）、MAPK/ERK激酶激酶1（MEKK 1）和Cullin 4-damage-specific DNA-binding protein 1（CUL 4-DDB 1）。WDR 68影响多种多样的生理功能，例如控制植物中的花青素合成、昆虫中的组织生长和脊椎动物中的颅面发育。然而，WDR 68活性的生化基础和调节机制在很大程度上仍然未知。为了更好地理解WDR 68的细胞功能，在这里，我们使用磷酸化蛋白质组学方法分离并鉴定了细胞WDR 68结合伴侣。超过200种具有广泛生物化学功能的细胞蛋白被鉴定为WDR 68结合蛋白候选物。八个T-复合物蛋白1（TCP 1）亚基组成的分子伴侣TCP 1环复合物/伴侣蛋白的TCP 1（TRiC/CCT）被确定为主要的WDR 68结合蛋白，并在WDR 68和TRiC/CCT的磷酸化位点进行了鉴定。免疫共沉淀实验证实了TRiC/CCT和WDR 68之间的结合。计算机辅助结构分析表明，WDR 68形成了一个七叶片β螺旋桨环。用一系列缺失突变体结合结构建模的实验表明，WDR 68的七个β-螺旋桨叶片中的三个对于TRiC/CCT结合是必需的和足够的。通过siRNA敲低细胞TRiC/CCT引起异常的WDR 68结构，并导致其DYRK 1A结合活性降低。同时，TRiC/CCT的敲低抑制了WDR 68的核积累，并且当在TRiC/CCT缺陷细胞中过表达时，WDR 68形成细胞聚集体。总之，我们的研究结果表明，分子伴侣TRiC/CCT是必不可少的正确的蛋白质折叠，DYRK 1A的结合，和核积累的WDR 68。
Background: Trp-Asp (WD) repeat protein 68 (WDR68) is a binding partner of dual specificity tyrosine phosphorylation-regulated protein kinase (DYRK1A), a kinase partly responsible for aspects of Down syndrome. Results: The molecular chaperone T-complex protein 1 (TCP1) ring complex/chaperonin-containing TCP1 (TRiC/CCT) binds to WDR68 and modulates its structure, DYRK1A-binding, and cellular localization. Conclusion: TRiC/CCT is essential for correct folding and function of WDR68. Significance: TRiC/CCT may have a general role in forming the functional WD40 repeat seven-bladed β-propeller structure. Trp-Asp (WD) repeat protein 68 (WDR68) is an evolutionarily conserved WD40 repeat protein that binds to several proteins, including dual specificity tyrosine phosphorylation-regulated protein kinase (DYRK1A), MAPK/ERK kinase kinase 1 (MEKK1), and Cullin4-damage-specific DNA-binding protein 1 (CUL4-DDB1). WDR68 affects multiple and diverse physiological functions, such as controlling anthocyanin synthesis in plants, tissue growth in insects, and craniofacial development in vertebrates. However, the biochemical basis and the regulatory mechanism of WDR68 activity remain largely unknown. To better understand the cellular function of WDR68, here we have isolated and identified cellular WDR68 binding partners using a phosphoproteomic approach. More than 200 cellular proteins with wide varieties of biochemical functions were identified as WDR68-binding protein candidates. Eight T-complex protein 1 (TCP1) subunits comprising the molecular chaperone TCP1 ring complex/chaperonin-containing TCP1 (TRiC/CCT) were identified as major WDR68-binding proteins, and phosphorylation sites in both WDR68 and TRiC/CCT were identified. Co-immunoprecipitation experiments confirmed the binding between TRiC/CCT and WDR68. Computer-aided structural analysis suggested that WDR68 forms a seven-bladed β-propeller ring. Experiments with a series of deletion mutants in combination with the structural modeling showed that three of the seven β-propeller blades of WDR68 are essential and sufficient for TRiC/CCT binding. Knockdown of cellular TRiC/CCT by siRNA caused an abnormal WDR68 structure and led to reduction of its DYRK1A-binding activity. Concomitantly, nuclear accumulation of WDR68 was suppressed by the knockdown of TRiC/CCT, and WDR68 formed cellular aggregates when overexpressed in the TRiC/CCT-deficient cells. Altogether, our results demonstrate that the molecular chaperone TRiC/CCT is essential for correct protein folding, DYRK1A binding, and nuclear accumulation of WDR68.
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影响因子：
16
作者：
Christoph Spiess;Erik J. Miller;A. J. McClellan;J. Frydman
通讯作者：
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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
发表时间：
2006
期刊：
The international journal of biochemistry & cell biology
影响因子：
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作者：
Yi,Chunling;Li,Shuting;Wang,Jian;Wei,Ning;Deng,XingWang
通讯作者：
Deng,XingWang
DOI：
10.1016/j.bbamcr.2011.07.018
发表时间：
2012-03
影响因子：
5.1
作者：
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通讯作者：
Neckers, Len
影响因子：
16
作者：
Fang, GW;Yu, HT;Kirschner, MW
通讯作者：
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影响因子：
16
作者：
Lu, ZM;Xu, SC;Hunter, T
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