TRIM4 interacts with TRPM8 and regulates its channel function through K423-mediated ubiquitination
TRIM4 interacts with TRPM8 and regulates its channel function through K423-mediated ubiquitination
复制标题
TRIM4 与 TRPM8 相互作用，并通过 K423 介导的泛素化调节其通道功能。
DOI：
10.1002/jcp.30065
复制
发表时间：
2020-10-09
影响因子：
5.6
通讯作者：
Tang, Jingfeng
中科院分区：
文献类型：
--
作者：
Huang, Yuan;Li, Shunyao;Tang, Jingfeng
瞬时受体电位美拉抑素成员8 （TRPM8）是一种Ca2+可渗透的非选择性阳离子通道，可被寒冷和冷却剂激活，介导异位性疼痛。在几种人类癌症中发现了TRPM8的功能障碍或异常表达。泛素化在TRPM8功能调控中的作用尚不清楚。在这里，我们确定了泛素(Ub)-连接酶E3， tripartite motif-containing 4 (TRIM4)，作为TRPM8的一个新的相互作用伙伴，并证实了TRIM4-TRPM8的相互作用是通过TRIM4的SPRY结构域介导的。膜片钳实验显示TRIM4负向调节HEK293细胞中trpm8介导的电流。此外，TRIM4通过促进TRPM8的k63连锁泛素化，降低了TRPM8在细胞表面的表达。进一步分析发现，TRPM8 n端赖氨酸残基423是TRIM4介导其功能调控的主要泛素化位点。ub激活酶E1， ub样修饰物激活酶1 （UBA1）也被发现与TRPM8相互作用，从而调节其通道功能和泛素化。此外，UBA1的敲低会破坏TRIM4对TRPM8泛素化和功能的调节。因此，本研究表明TRIM4通过k423介导的TRPM8泛素化下调TRPM8，并需要UBA1调控TRPM8。
Transient receptor potential melastatin member 8 (TRPM8), a Ca2+-permeable nonselective cation channel activated by cold and cooling agents, mediates allodynia. Dysfunction or abnormal expression of TRPM8 has been found in several human cancers. The role of ubiquitination in the regulation of TRPM8 function remains poorly understood. Here, we identified the ubiquitin (Ub)-ligase E3, tripartite motif-containing 4 (TRIM4), as a novel interaction partner of TRPM8 and confirmed that the TRIM4-TRPM8 interaction was mediated through the SPRY domain of TRIM4. Patch-clamp assays showed that TRIM4 negatively regulates TRPM8-mediated currents in HEK293 cells. Moreover, TRIM4 reduced the expression of TRPM8 on the cell surface by promoting the K63-linked ubiquitination of TRPM8. Further analyses revealed that the TRPM8 N-terminal lysine residue at 423 was the major ubiquitination site that mediates its functional regulation by TRIM4. A Ub-activating enzyme E1, Ub-like modifier-activating enzyme 1 (UBA1), was also found to interact with TRPM8, thereby regulating its channel function and ubiquitination. In addition, knockdown of UBA1 impaired the regulation of TRPM8 ubiquitination and function by TRIM4. Thus, this study demonstrates that TRIM4 downregulates TRPM8 via K423-mediated TRPM8 ubiquitination and requires UBA1 to regulate TRPM8.