Evidence of LAT as a dual substrate for Lck and Syk in T lymphocytes.
LAT is a linker protein essential for activation of T lymphocytes. Its rapid tyrosine-phosphorylation upon T cell receptor (TCR) stimulation recruits downstream signaling molecules for membrane targeting and activation. LAT is physically concentrated in cholesterol-enriched membrane microdomains and is known a substrate for Syk/Zap70 kinase. In this study, we demonstrate that LAT serves as a dual substrate for both Lck and Syk kinases. LAT phosphorylation is absent in Lck-deficient J.CaM1.6 cells and Lck is co-precipitated with LAT in pervanadate-activated Jurkat cells. Further, the in vitro kinase assay using purified Lck and LAT shows that Lck directly phosphorylates LAT. Both Lck and Syk, phosphorylate the ITAM-like motifs on LAT at Y171Y191, which is essential for induction of the interaction of LAT with downstream signaling molecules such as Grb2, PLC-gamma1 and c-Cbl, and for activation of MAPK-ERK. Collectively, our data indicate that LAT is an immediate substrate for Lck in one of the earliest events of T cell activation.
Jiang
Yixing
Y
Department of Medicine and Pennstate Cancer Institute, Penn State University College of Medicine, Hershey, PA 17033, United States.
Cheng
Hua
H
D048868:Adaptor Proteins, Signal Transducing / Q000378:metabolism*; D002478:Cells, Cultured; D051380:GRB2 Adaptor Protein / Q000378:metabolism; D006801:Humans; D047908:Intracellular Signaling Peptides and Proteins / Q000378:metabolism*; D019169:Jurkat Cells / Q000378:metabolism; D007668:Kidney / Q000166:cytology / Q000378:metabolism; D008213:Lymphocyte Activation; D019860:Lymphocyte Specific Protein Tyrosine Kinase p56(lck) / Q000378:metabolism*; D008565:Membrane Proteins / Q000378:metabolism*; D019950:Mitogen-Activated Protein Kinase 1 / Q000378:metabolism; D048052:Mitogen-Activated Protein Kinase 3 / Q000378:metabolism; D020928:Mitogen-Activated Protein Kinases / Q000378:metabolism; D051966:Phospholipase C gamma / Q000378:metabolism; D010766:Phosphorylation; D011505:Protein-Tyrosine Kinases / Q000378:metabolism*; D050721:Proto-Oncogene Proteins c-cbl / Q000378:metabolism; D015398:Signal Transduction; D000072377:Syk Kinase; D013601:T-Lymphocytes / Q000378:metabolism*
D048868:Adaptor Proteins, Signal Transducing; D051380:GRB2 Adaptor Protein; D047908:Intracellular Signaling Peptides and Proteins; C110810:LAT protein, human; D008565:Membrane Proteins; D050721:Proto-Oncogene Proteins c-cbl; D011505:Protein-Tyrosine Kinases; D019860:Lymphocyte Specific Protein Tyrosine Kinase p56(lck); C000606896:SYK protein, human; D000072377:Syk Kinase; D019950:Mitogen-Activated Protein Kinase 1; D048052:Mitogen-Activated Protein Kinase 3; D020928:Mitogen-Activated Protein Kinases; D051966:Phospholipase C gamma; C067419:CBL protein, human