Lee, Dong Kun Park, Jung Yeon Yoo, Jae Cheal Byun, Eun Hye Bae, Yeon-Ju Lee, Young-Sun Park, Nammi Kang, Dawon Han, Jaehee Park, Jae Yong Hwang, Eunmi Hong, Seong-Geun 2024-01-19T21:33:41Z 2024-01-19T21:33:41Z 2021-09-05 2018-10 0031-6768 https://pubs.kist.re.kr/handle/201004/120832 Transient receptor-potential, cation channel, subfamily M, member 4 (TRPM4) channels regulate a variety of physiological and pathological processes; however, their roles as functional channels under diverse conditions remain unclear. In this study, cytosolic protein tyrosine phosphatase non-receptor type 6 (PTPN6) interacted with TRPM4 channels. We confirmed their interaction by performing co-immunoprecipitation (Co-IP) assays following heterologous PTPN6 and TRPM4 channel expression in HEK293 cells. Furthermore, biomolecular fluorescence complementation (BiFC) image analysis confirmed TRPM4-PTPN6 binding. In addition, immunoblotting and Co-IP analyses revealed that TRPM4 expression significantly decreased in the membrane fraction of cells after PTPN6 was silenced with a specific short-hairpin RNA (shRNA-PTPN6). In agreement, TRPM4-induced changes in whole-cell currents were not detected in PTPN6-silenced HEK cells, in contrast to cells transfected with a scrambled RNA (scRNA) or in naive HEK cells. These data suggest that PTPN6 inhibits TRPM4 channel activity by disrupting TRPM4 expression. Furthermore, TRPM4 channels were expressed in the membrane of naive cells and scRNA transfectants, but not in those of PTPN6-silenced cells. These results indicated that PTPN6 is critically associated with TRPM4 trafficking. This role of PTPN6 in TRPM4 membrane localization was also demonstrated in HeLa cells. TRPM4 overexpression significantly enhanced cell proliferation in untreated HeLa cells, but not in HeLa cells with silenced PTPN6 expression. These findings indicate that PTPN6-dependent TRPM4 expression and trafficking to the plasma membrane is critical for cell proliferation in both HEK293 and HeLa cells. Therefore, PTPN6 is a novel therapeutic target for treating pathologic diseases involving TRPM4. English SPRINGER HEIDELBERG NONSELECTIVE CATION CHANNEL POTENTIAL MELASTATIN 4 TYROSINE PHOSPHATASES PROTEIN SHP-1 PTPN6 regulates the cell-surface expression of TRPM4 channels in HEK293 cells Article 10.1007/s00424-018-2161-9 1 PFLUGERS ARCHIV-EUROPEAN JOURNAL OF PHYSIOLOGY, v.470, no.10, pp.1449 - 1458 PFLUGERS ARCHIV-EUROPEAN JOURNAL OF PHYSIOLOGY 470 10 1449 1458 scie scopus 000444576600003 2-s2.0-85048775037 Physiology Physiology Article NONSELECTIVE CATION CHANNEL POTENTIAL MELASTATIN 4 TYROSINE PHOSPHATASES PROTEIN SHP-1 TRPM4 PTPN6 RTK Tyrosine phosphorylation Membrane trafficking