Up-regulation of the IKCa1 potassium channel during T-cell activation. Molecular mechanism and functional consequences.
We used whole cell recording to evaluate functional expression of the intermediate conductance Ca(2+)-activated K(+) channel, IKCa1, in response to various mitogenic stimuli. One to two days following engagement of T-cell receptors to trigger both PKC- and Ca(2+)-dependent events, IKCa1 expression increased from an average of 8 to 300-800 channels/cell. Selective stimulation of the PKC pathway resulted in equivalent up-regulation, whereas a calcium ionophore was relatively ineffective. Enhancement in IKCa1 mRNA levels paralleled the increased channel number. The genomic organization of IKCa1, SKCa2, and SKCa3 were defined, and IK(Ca) and SK(Ca) genes were found to have a remarkably similar intron-exon structure. Mitogens enhanced IKCa1 promoter activity proportional to the increase in IKCa1 mRNA, suggesting that transcriptional mechanisms underlie channel up-regulation. Mutation of motifs for AP1 and Ikaros-2 in the promoter abolished this induction. Selective Kv1.3 inhibitors ShK-Dap(22), margatoxin, and correolide suppressed mitogenesis of resting T-cells but not preactivated T-cells with up-regulated IKCa1 channel expression. Selectively blocking IKCa1 channels with clotrimazole or TRAM-34 suppressed mitogenesis of preactivated lymphocytes, whereas resting T-cells were less sensitive. Thus, Kv1.3 channels are essential for activation of quiescent cells, but signaling through the PKC pathway enhances expression of IKCa1 channels that are required for continued proliferation.
Ghanshani
S
S
Department of Physiology and Biophysics, and Department of Microbiology and Molecular Genetics, University of California, Irvine, California 92697, USA.
Wulff
H
H
Miller
M J
MJ
Rohm
H
H
Neben
A
A
Gutman
G A
GA
Cahalan
M D
MD
Chandy
K G
KG
D002121:Calcium Channel Blockers / Q000494:pharmacology; D015220:Calcium Channels / Q000096:biosynthesis* / Q000235:genetics; D006801:Humans; D051660:Intermediate-Conductance Calcium-Activated Potassium Channels; D008213:Lymphocyte Activation / Q000187:drug effects; D008934:Mitogens / Q000494:pharmacology; D008954:Models, Biological; D008969:Molecular Sequence Data; D010835:Phytohemagglutinins / Q000494:pharmacology; D015221:Potassium Channels; D011401:Promoter Regions, Genetic; D011720:Pyrazoles / Q000494:pharmacology; D015398:Signal Transduction; D013601:T-Lymphocytes / Q000187:drug effects / Q000276:immunology / Q000378:metabolism*; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology; D014158:Transcription, Genetic; D015854:Up-Regulation
D002121:Calcium Channel Blockers; D015220:Calcium Channels; D051660:Intermediate-Conductance Calcium-Activated Potassium Channels; C496923:KCNN4 protein, human; D008934:Mitogens; D010835:Phytohemagglutinins; D015221:Potassium Channels; D011720:Pyrazoles; C411671:TRAM 34; D013755:Tetradecanoylphorbol Acetate