Human p59fyn(T) regulates OKT3-induced calcium influx by a mechanism distinct from PIP2 hydrolysis in Jurkat T cells.
The earliest biochemical event after cross-linking of TCR is the tyrosine phosphorylation of a variety of substrates. At least three nonreceptor tyrosine kinases have been implicated in this signaling cascade: p59fyn(T), p56lck, and ZAP-70. Recently, PLC gamma 1 has been shown to be tyrosine phosphorylated in T cells after receptor activation. This increase in tyrosine phosphorylation correlates with the increased activity of the enzyme. The substrate for PLC gamma 1, phosphatidylinositol 4,5-bisphosphate (PIP2), is hydrolyzed to the protein kinase C activator diacylglycerol and inositol 1,4,5-triphosphate (IP3), which promotes calcium release from the endoplasmic reticulum. These results lend support to the notion that calcium mobilization after TCR cross-linking is mediated by increased levels of IP3. In this study we have cloned and transfected a human p59fyn(T) cDNA in the anti-sense configuration into the human T cell line, Jurkat, resulting in decreased expression of the protein. We find that cell lines expressing significantly reduced levels of p59fyn(T) exhibit significantly lower calcium influx following OKT3 activation. However, the level of IP3 production was unchanged and IP1 and IP2 levels were elevated. These data indicate that p59fyn(T) can regulate calcium influx by a mechanism distinct from PIP2 hydrolysis.
Rigley
K
K
Cantab Pharmaceuticals Research Limited, Cambridge, UK.
Slocombe
P
P
Proudfoot
K
K
Wahid
S
S
Mandair
K
K
Bebbington
C
C
D000595:Amino Acid Sequence; D001483:Base Sequence; D002118:Calcium / Q000378:metabolism*; D002460:Cell Line; D018076:DNA, Complementary / Q000032:analysis; D006801:Humans; D006868:Hydrolysis; D008969:Molecular Sequence Data; D016853:Muromonab-CD3 / Q000276:immunology; D018129:Phosphatidylinositol Phosphates / Q000378:metabolism*; D011505:Protein-Tyrosine Kinases / Q000502:physiology*; D011518:Proto-Oncogene Proteins / Q000502:physiology*; D051076:Proto-Oncogene Proteins c-fyn; D016372:RNA, Antisense / Q000096:biosynthesis; D011948:Receptors, Antigen, T-Cell / Q000276:immunology; D013601:T-Lymphocytes / Q000378:metabolism*
D018076:DNA, Complementary; D016853:Muromonab-CD3; D018129:Phosphatidylinositol Phosphates; D011518:Proto-Oncogene Proteins; D016372:RNA, Antisense; D011948:Receptors, Antigen, T-Cell; D011505:Protein-Tyrosine Kinases; C494957:FYN protein, human; D051076:Proto-Oncogene Proteins c-fyn; D002118:Calcium