Differential regulation of p72syk expression in naive and proliferating CD4-CD8+ T cells.
A well-known consequence of TCR stimulation in proliferating T cells is cell death by apoptosis. We have previously shown that the extent of tyrosine phosphorylation of TCR zeta, CD3 gamma, and CD3 epsilon subunits in proliferating CD4-CD8+ T cells after TCR stimulation was decreased when compared to similarly stimulated naive T cells expressing the same TCR. Furthermore, these differences correlated with a decrease in the specific kinase activity of p56lck and p59fyn, with a corresponding increase in the specific kinase activity of p50rsk, a negative regulator of src-family tyrosine kinases. In this study we determined whether kinases that bind tyrosine phosphorylated TCR zeta chain were differentially regulated in naive and proliferating cells. Chemically synthesized cytoplasmic domains of the TCR zeta chain were fully phosphorylated in vitro with p56lck and used to precipitate TCR zeta binding proteins in naive and proliferating cells. Using this method we found that both ZAP-70 and p72syk bound tyrosine phosphorylated TCR zeta very efficiently. More interestingly, p72syk was found to be expressed only in naive but not proliferating cells. Kinetic studies indicate that more than 48 hr of activation was required for ceasation of p72syk expression. We also showed that the inability to detect p72syk expression in proliferating cells was not due to its translocation to cytoskeletal compartments in proliferating cells. We propose that the differential regulation of ZAP-70 and p72syk in naive and proliferating cells may contribute to the uncoupling of the TCR signaling pathway from downstream signaling events leading to distinct functional outcomes in these two cell types after TCR stimulation.
Orchansky
P L
PL
Department of Microbiology and Immunology, University of British Columbia, Vancouver, Canada.
Matsuuchi
L
L
Ericsson
P O
PO
Teh
H S
HS
D000818:Animals; D015153:Blotting, Western; D017252:CD3 Complex / Q000378:metabolism; D018414:CD8-Positive T-Lymphocytes / Q000378:metabolism*; D002850:Chromatography, Gel; D004792:Enzyme Precursors / Q000096:biosynthesis*; D006801:Humans; D047908:Intracellular Signaling Peptides and Proteins; D019169:Jurkat Cells; D008213:Lymphocyte Activation; D008565:Membrane Proteins / Q000378:metabolism; D051379:Mice; D008822:Mice, Transgenic; D010766:Phosphorylation; D011505:Protein-Tyrosine Kinases / Q000096:biosynthesis* / Q000378:metabolism; D011948:Receptors, Antigen, T-Cell / Q000378:metabolism; D012685:Sepharose; D015398:Signal Transduction; D000072377:Syk Kinase; D051746:ZAP-70 Protein-Tyrosine Kinase
D017252:CD3 Complex; D004792:Enzyme Precursors; D047908:Intracellular Signaling Peptides and Proteins; D008565:Membrane Proteins; D011948:Receptors, Antigen, T-Cell; C054979:antigen T cell receptor, zeta chain; D012685:Sepharose; D011505:Protein-Tyrosine Kinases; C000606896:SYK protein, human; D000072377:Syk Kinase; C000606897:Syk protein, mouse; D051746:ZAP-70 Protein-Tyrosine Kinase; C497261:ZAP70 protein, human; C497262:Zap70 protein, mouse