The B-box module of CYLD is responsible for its intermolecular interaction and cytoplasmic localization.
The B-box module of CYLD is responsible for its intermolecular interaction and cytoplasmic localization.
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CYLD的B-box模块负责其分子间相互作用和细胞质定位
DOI：
10.18632/oncotarget.15142
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发表时间：
2017-08-01
期刊：
影响因子：
--
通讯作者：
Liu M
中科院分区：
文献类型：
--
作者：
Xie S;Chen M;Gao S;Zhong T;Zhou P;Li D;Zhou J;Gao J;Liu M
肿瘤抑制蛋白圆柱瘤病（CYLD）是一种微管相关的去泛素化酶，在天然免疫、细胞分裂和纤毛发生等细胞活动中发挥重要作用。结构表征揭示了一个小的锌结合B盒插入在泛素特异性蛋白酶（USP）结构域的CYLD;然而，这个模块的确切作用仍有待阐明。在这里，我们确定了一个关键的作用，B盒在促进分子间的相互作用和亚细胞定位的CYLD。通过免疫共沉淀试验，我们发现CYLD具有形成分子间复合物的能力。天然凝胶电泳分析和下拉分析表明，USP结构域的CYLD是必不可少的分子间相互作用。进一步的研究表明，从USP结构域中删除B盒破坏了CYLD的分子间相互作用。重要的是，虽然B盒的丢失对CYLD的去泛素化酶活性没有明显影响，但它消除了USP结构域介导的CYLD在细胞质中的保留。这些数据表明CYLD的B-box模块在介导其组装和亚细胞分布中具有重要作用，这可能与CYLD在各种生物过程中的功能有关。
The tumor suppressor protein cylindromatosis (CYLD), as a microtubule-associated deubiquitinase, plays a pivotal role in a wide range of cellular activities, including innate immunity, cell division, and ciliogenesis. Structural characterization reveals a small zinc-binding B-box inserted within the ubiquitin specific protease (USP) domain of CYLD; however, the exact role for this module remains yet to be elucidated. Here we identify a critical role for the B-box in facilitating the intermolecular interaction and subcellular localization of CYLD. By co-immunoprecipitation assays we uncover that CYLD has the ability to form an intermolecular complex. Native gel electrophoresis analysis and pull down assays show that the USP domain of CYLD is essential for its intermolecular interaction. Further investigation reveals that deletion of the B-box from the USP domain disrupts the intermolecular interaction of CYLD. Importantly, although loss of the B-box has no obvious effect on the deubiquitinase activity of CYLD, it abolishes the USP domain-mediated retention of CYLD in the cytoplasm. Collectively, these data demonstrate an important role for the B-box module of CYLD in mediating its assembly and subcellular distribution, which might be related to the functions of CYLD in various biological processes.