The juvenile myoclonic epilepsy-related protein EFHC1 interacts with the redox-sensitive TRPM2 channel linked to cell death.
The transient receptor potential M2 channel (TRPM2) is the Ca(2+)-permeable cation channel controlled by cellular redox status via β-NAD(+) and ADP-ribose (ADPR). TRPM2 activity has been reported to underlie susceptibility to cell death and biological processes such as inflammatory cell migration and insulin secretion. However, little is known about the intracellular mechanisms that regulate oxidative stress-induced cell death via TRPM2. We report here a molecular and functional interaction between the TRPM2 channel and EF-hand motif-containing protein EFHC1, whose mutation causes juvenile myoclonic epilepsy (JME) via mechanisms including neuronal apoptosis. In situ hybridization analysis demonstrates TRPM2 and EFHC1 are coexpressed in hippocampal neurons and ventricle cells, while immunoprecipitation analysis demonstrates physical interaction of the N- and C-terminal cytoplasmic regions of TRPM2 with the EFHC1 protein. Coexpression of EFHC1 significantly potentiates hydrogen peroxide (H(2)O(2))- and ADPR-induced Ca(2+) responses and cationic currents via recombinant TRPM2 in HEK293 cells. Furthermore, EFHC1 enhances TRPM2-conferred susceptibility of HEK293 cells to H(2)O(2)-induced cell death, which is reversed by JME mutations. These results reveal a positive regulatory action of EFHC1 on TRPM2 activity, suggesting that TRPM2 contributes to the expression of JME phenotypes by mediating disruptive effects of JME mutations of EFHC1 on biological processes including cell death.
Laboratory of Molecular Biology, Department of Synthetic Chemistry and Biological Chemistry, Graduate School of Engineering, Kyoto University, Kyoto, Japan.
Masahiro
M
Katano
Tomohiro
T
Numata
Kripamoy
K
Aguan
Yuji
Y
Hara
Shigeki
S
Kiyonaka
Shinichiro
S
Yamamoto
Takafumi
T
Miki
Seishiro
S
Sawamura
Toshimitsu
T
Suzuki
Kazuhiro
K
Yamakawa
Yasuo
Y
Mori
D000246:Adenosine Diphosphate Ribose / Q000235:genetics / Q000378:metabolism; D020816:Amino Acid Motifs; D000818:Animals; D017209:Apoptosis; D002135:Calcium-Binding Proteins / Q000235:genetics / Q000378:metabolism*; D005786:Gene Expression Regulation / Q000187:drug effects / Q000235:genetics; D057809:HEK293 Cells; D006624:Hippocampus / Q000378:metabolism / Q000473:pathology; D006801:Humans; D006861:Hydrogen Peroxide / Q000494:pharmacology; D051379:Mice; D009154:Mutation; D020190:Myoclonic Epilepsy, Juvenile / Q000235:genetics / Q000378:metabolism / Q000473:pathology; D009243:NAD / Q000235:genetics / Q000378:metabolism; D009474:Neurons / Q000378:metabolism / Q000473:pathology; D016877:Oxidants / Q000494:pharmacology; D017434:Protein Structure, Tertiary; D050053:TRPM Cation Channels / Q000235:genetics / Q000378:metabolism*
D002135:Calcium-Binding Proteins; C489648:EFHC1 protein, human; C520734:EFHC1 protein, mouse; D016877:Oxidants; D050053:TRPM Cation Channels; C121934:TRPM2 protein, human; C544443:TRPM2 protein, mouse; D009243:NAD; D000246:Adenosine Diphosphate Ribose; D006861:Hydrogen Peroxide