Sam68 association with p120GAP in CD4+ T cells is dependent on CD4 molecule expression.
p120 GTPase-activating protein (p120GAP) is a major negative regulator of p21ras activity in several cell types including T cells. Catalytic activity of this enzyme is regulated in part by its interaction with several associated tyrosine-phosphorylated proteins. Sam68 was initially described as associated with p120GAP. It has been further established that Sam68 is a substrate of src kinases in mitosis and that it is not associated with p120GAP in transformed fibroblasts. We describe herein that Sam68 associates with p120GAP and PLC gamma 1 in human mature T cells and in a T cell line expressing the CD4 molecule HUT78 CD4+. This association is present in nonactivated cells and increases after anti-CD3 activation. It is dependent on CD4 expression and, in part, on the association of CD4 with p56lck, as shown by the strongly decreased association of Sam68 with p120GAP in the CD4- mutants, HUT78 CD4-, and by the reduced association of Sam68 with both p120GAP and p56lck in the HUT78 T cell line expressing a CD4 mutant unable to interact with p56lck, HUT78 C420/22. We propose that recruitment of Sam68, via CD4/p56lck, to the inner face of the plasma membrane may permit, via its docking properties, the correct association of key signaling molecules including PLC gamma 1 and p120GAP. This formation of transduction modules will enable the activation of different signaling cascades including the p21ras pathway and an array of downstream events, ultimately leading to T cell activation.
Jabado
N
N
Institut National de la Santé et de la Recherche Médicale, Unité 429, Hôpital Necker-Enfants Malades, Paris, France. Jabado@necker.fr
Jauliac
S
S
Pallier
A
A
Bernard
F
F
Fischer
A
A
Hivroz
C
C
D048868:Adaptor Proteins, Signal Transducing; D015704:CD4 Antigens / Q000096:biosynthesis* / Q000502:physiology; D015496:CD4-Positive T-Lymphocytes / Q000201:enzymology / Q000276:immunology / Q000378:metabolism*; D002478:Cells, Cultured; D004268:DNA-Binding Proteins; D020558:GTP Phosphohydrolases / Q000378:metabolism*; D020690:GTPase-Activating Proteins; D006801:Humans; D007527:Isoenzymes / Q000378:metabolism; D008213:Lymphocyte Activation; D019860:Lymphocyte Specific Protein Tyrosine Kinase p56(lck) / Q000378:metabolism / Q000502:physiology; D016399:Lymphoma, T-Cell; D051966:Phospholipase C gamma; D010766:Phosphorylation; D011506:Proteins / Q000378:metabolism*; D016601:RNA-Binding Proteins / Q000378:metabolism*; D014407:Tumor Cells, Cultured; D010738:Type C Phospholipases / Q000378:metabolism; D020703:ras GTPase-Activating Proteins; D018631:ras Proteins / Q000378:metabolism*; D019061:src-Family Kinases / Q000378:metabolism*
D048868:Adaptor Proteins, Signal Transducing; D015704:CD4 Antigens; D004268:DNA-Binding Proteins; D020690:GTPase-Activating Proteins; D007527:Isoenzymes; C489559:KHDRBS1 protein, human; D011506:Proteins; D016601:RNA-Binding Proteins; D020703:ras GTPase-Activating Proteins; D019860:Lymphocyte Specific Protein Tyrosine Kinase p56(lck); D019061:src-Family Kinases; D010738:Type C Phospholipases; D051966:Phospholipase C gamma; D020558:GTP Phosphohydrolases; D018631:ras Proteins