A novel TRPM2 isoform inhibits calcium influx and susceptibility to cell death.
TRPM2 is a Ca(2+)-permeable channel that is activated by oxidative stress and confers susceptibility to cell death. Here, an isoform of TRPM2 was identified in normal human bone marrow that consists of the TRPM2 N terminus and the first two predicted transmembrane domains. Because of alternative splicing, a stop codon (TAG) is located at the splice junction between exons 16 and 17, resulting in deletion of the four C-terminal transmembrane domains, the putative calcium-permeable pore region, and the entire C terminus. This splice variant was found in other hematopoietic cells including human burst forming unit-erythroid-derived erythroblasts and TF-1 erythroleukemia cells. Endogenous expression of both the short form of TRPM2 (TRPM2-S) and the full length (TRPM2-L) was determined by reverse transcriptase-PCR, and localization of endogenous TRPM2 to the plasma membrane was demonstrated by confocal microscopy. Heterologous expression of TRPM2-S in HEK 293T cells demonstrated similar membrane localization as TRPM2-L, and coexpression of TRPM2-S did not alter the subcellular localization of TRPM2-L. The direct interaction of TRPM2-S with TRPM2-L was demonstrated with immunoprecipitation. H(2)O(2) induced calcium influx through TRPM2-L expressed in 293T cells. Coexpression of TRPM2-S suppressed H(2)O(2)-induced calcium influx through TRPM2-L. Furthermore, expression of TRPM2-S inhibited susceptibility to cell death and onset of apoptosis induced by H(2)O(2) in cells expressing TRPM2-L. These data demonstrate that TRPM2-S is an important physiologic isoform of TRPM2 and modulates channel activity and induction of cell death by oxidative stress through TRPM2-L.
Zhang
Wenyi
W
Henry Hood Research Program, Sigfried and Janet Weis Center for Research, The Geisinger Clinic, 100 North Academy Avenue, Danville, PA 17822, USA.
Chu
Xin
X
Tong
Qin
Q
Cheung
Joseph Y
JY
Conrad
Kathleen
K
Masker
Kathryn
K
Miller
Barbara A
BA
D017209:Apoptosis; D002118:Calcium / Q000378:metabolism*; D015220:Calcium Channels / Q000032:analysis / Q000502:physiology*; D002460:Cell Line; D003001:Cloning, Molecular; D006801:Humans; D006861:Hydrogen Peroxide / Q000494:pharmacology; D007473:Ion Channels; D008565:Membrane Proteins; D020033:Protein Isoforms; D050053:TRPM Cation Channels
D015220:Calcium Channels; D007473:Ion Channels; D008565:Membrane Proteins; D020033:Protein Isoforms; D050053:TRPM Cation Channels; C121934:TRPM2 protein, human; D006861:Hydrogen Peroxide; D002118:Calcium