Premium
TRIM4 interacts with TRPM8 and regulates its channel function through K423‐mediated ubiquitination
Author(s) -
Huang Yuan,
Li Shunyao,
Jia Zhenhua,
Li Shi,
He Wenzao,
Zhou Cefan,
Zhang Rui,
Xu Rui,
Sun Bo,
Ali Declan William,
Michalak Marek,
Chen XingZhen,
Tang Jingfeng
Publication year - 2021
Publication title -
journal of cellular physiology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.529
H-Index - 174
eISSN - 1097-4652
pISSN - 0021-9541
DOI - 10.1002/jcp.30065
Subject(s) - trpm8 , ubiquitin , ubiquitin ligase , transient receptor potential channel , hek 293 cells , microbiology and biotechnology , chemistry , gene knockdown , biochemistry , receptor , biology , gene , trpv1
Abstract Transient receptor potential melastatin member 8 (TRPM8), a Ca 2+ ‐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.