The interacting domains in cereblon differentially modulate the immunomodulatory drug-mediated ubiquitination and degradation of its binding partners
The interacting domains in cereblon differentially modulate the immunomodulatory drug-mediated ubiquitination and degradation of its binding partners
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cereblon 中的相互作用结构域差异调节免疫调节药物介导的泛素化及其结合伴侣的降解
DOI：
10.1016/j.bbrc.2018.11.058
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发表时间：
2018
期刊：
影响因子：
--
通讯作者：
Guoqiang Xu
中科院分区：
文献类型：
--
作者：
Jing Tao;Jing Yang;Guoqiang Xu
Cereblon（CRBN）是cullin-4 RING E3连接酶（CRL 4）的底物受体，已被用于通过小分子量CRBN调节剂降解靶向蛋白。然而，目前还不清楚在不同结构域与CRBN相互作用的蛋白质是否以及如何受到这些调节剂的影响。在这里，我们使用CRBN和它的四个结合伙伴，c-Jun，氯离子通道蛋白CLC-1，转录因子IKZF 1和MEIS 2，作为模型蛋白质，研究免疫调节药物（IMiDs），包括沙利度胺，来那度胺和泊马度胺，对它们的稳定性，泛素化和与CRBN的相互作用的影响。结合先前的发现，结构域作图实验表明，这四种蛋白质与CRBN在三个不同的区域相互作用。免疫印迹分析表明，CRBN结合伴侣的蛋白水平可以增强，减弱，或不受IMiDs的影响。相互作用分析和泛素化分析表明，IMiDs调节CRBN和它的结合伙伴之间的相互作用，在三个不同的方式，从而差异调节其泛素化。这项工作表明CRBN中的结合结构域是影响IMiDs对这些CRBN相互作用伴侣的泛素化和稳定性的调节的关键因素。
Cereblon (CRBN), a substrate receptor of the cullin-4 RING E3 ligase (CRL4), has been utilized for the targeted protein degradation via small molecular weight CRBN modulators. However, it is unclear whether and how proteins that interact with CRBN at different domains are affected by these modulators. Here, we use CRBN and its four binding partners, c-Jun, chloride channel protein CLC-1, transcription factor IKZF1, and MEIS2, as model proteins to investigate the effect of immunomodulatory drugs (IMiDs) including thalidomide, lenalidomide, and pomalidomide, on their stability, ubiquitination, and interaction with CRBN. Together with previous discoveries, domain mapping experiment shows that these four proteins interact with CRBN at three distinct regions. Immunoblotting analyses reveal that the protein level of CRBN-binding partners could be enhanced, attenuated, or not affected by IMiDs. Interaction analyses and ubiquitination assay demonstrate that IMiDs modulate the interaction between CRBN and its binding partners in three distinct ways and thus differentially regulate their ubiquitination. This work suggests that the binding domain in CRBN is a critical factor which influences the regulation of IMiDs on the ubiquitination and stability of these CRBN-interacting partners.