Coupling Ca2+ store release to Icrac channel activation in B lymphocytes requires the activity of Lyn and Syk kinases.
Activation of the B cell receptor complex in B lymphocytes causes Ca(2+) release from intracellular stores, which, in turn, activates ion channels known as Icrac. We investigated the mechanisms that link Ca(2+) store release to channel gating in DT40 B lymphocyte cell lines genetically manipulated to suppress the expression of several tyrosine kinases: Btk, Lyn, Syk, and the Blnk adaptor molecule. The simultaneous but not the independent suppression of Lyn and Syk expression prevents the activation of Icrac without interfering with thapsigargin-sensitive Ca(2+) store release. Icrac activation by Ca(2+) is reversed in mutant cells by the homologous expression of the missing kinases. Pharmacological inhibition of kinase activity by LavendustinA and PP2 cause the same functional deficit as the genetic suppression of enzyme expression. Biochemical assays demonstrate that kinase activity is required as a tonic signal: targets must be phosphorylated to link Ca(2+) store release to Icrac gating. The action of kinases on Icrac activation does not arise from control of the expression level of the stromal interaction molecule 1 and Orai1 proteins.
Chung
S Clare
SC
Department of Physiology, School of Medicine, University of California, San Francisco, San Francisco, CA 94143, USA.
Limnander
Andre
A
Kurosaki
Tomohiro
T
Weiss
Arthur
A
Korenbrot
Juan I
JI
D000818:Animals; D001402:B-Lymphocytes / Q000378:metabolism*; D002118:Calcium / Q000378:metabolism*; D015220:Calcium Channels / Q000378:metabolism*; D045744:Cell Line, Tumor; D002645:Chickens; D004594:Electrophysiology; D047908:Intracellular Signaling Peptides and Proteins / Q000235:genetics / Q000378:metabolism*; D015640:Ion Channel Gating; D008213:Lymphocyte Activation; D008565:Membrane Proteins / Q000378:metabolism; D009154:Mutation; D010636:Phenols / Q000378:metabolism; D010766:Phosphorylation; D011505:Protein-Tyrosine Kinases / Q000235:genetics / Q000378:metabolism*; D012333:RNA, Messenger; D000072377:Syk Kinase; D019284:Thapsigargin / Q000494:pharmacology; D019061:src-Family Kinases / Q000235:genetics / Q000378:metabolism*
D015220:Calcium Channels; D047908:Intracellular Signaling Peptides and Proteins; D008565:Membrane Proteins; D010636:Phenols; D012333:RNA, Messenger; C062898:lavendustin A; D019284:Thapsigargin; D011505:Protein-Tyrosine Kinases; D000072377:Syk Kinase; C060902:lyn protein-tyrosine kinase; D019061:src-Family Kinases; D002118:Calcium