p62dok negatively regulates CD2 signaling in Jurkat cells.
p62(dok) belongs to a newly identified family of adaptor proteins. In T cells, the two members that are predominantly expressed, p56(dok) and p62(dok), are tyrosine phosphorylated upon CD2 or CD28 stimulation, but not upon CD3 ligation. Little is known about the biological role of Dok proteins in T cells. In this study, to evaluate the importance of p62(dok) in T cell function, we generated Jurkat clones overexpressing p62(dok). Our results demonstrate that overexpression of p62(dok) in Jurkat cells has a dramatic negative effect on CD2-mediated signaling. The p62(dok)-mediated inhibition affects several biochemical events initiated by CD2 ligation, such as the increase of intracellular Ca(2+), phospholipase C gamma 1 activation, and extracellular signal-regulated kinase 1/2 activation. Importantly, these cellular events are not affected in the signaling cascade induced by engagement of the CD3/TCR complex. However, both CD3- and CD2-induced NF-AT activation and IL-2 secretion are impaired in p62(dok)-overexpressing cells. In addition, we show that CD2 but not CD3 stimulation induces p62(dok) and Ras GTPase-activating protein recruitment to the plasma membrane. These results suggest that p62(dok) plays a negative role at multiple steps in the CD2 signaling pathway. We propose that p62(dok) may represent an important negative regulator in the modulation of the response mediated by the TCR.
Institut National de la Recherche Scientifique-Institut Armand-Frappier, Université du Québec, Laval QC, Canada.
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Némorin
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Laporte
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Bérubé
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Duplay
D018801:CD2 Antigens / Q000378:metabolism / Q000502:physiology*; D017252:CD3 Complex / Q000502:physiology; D002118:Calcium / Q000037:antagonists & inhibitors / Q000378:metabolism; D020013:Calcium Signaling / Q000235:genetics / Q000276:immunology; D002462:Cell Membrane / Q000276:immunology / Q000378:metabolism; D002999:Clone Cells; D004268:DNA-Binding Proteins; D015536:Down-Regulation / Q000235:genetics / Q000276:immunology*; D005786:Gene Expression Regulation / Q000276:immunology; D005822:Genetic Vectors / Q000378:metabolism; D006801:Humans; D007376:Interleukin-2 / Q000037:antagonists & inhibitors / Q000096:biosynthesis / Q000235:genetics; D019169:Jurkat Cells / Q000201:enzymology / Q000276:immunology* / Q000378:metabolism* / Q000502:physiology; D010750:Phosphoproteins / Q000096:biosynthesis / Q000235:genetics / Q000378:metabolism / Q000502:physiology*; D010766:Phosphorylation; D016601:RNA-Binding Proteins; D015398:Signal Transduction / Q000235:genetics / Q000276:immunology*; D014162:Transfection; D014443:Tyrosine / Q000378:metabolism; D020703:ras GTPase-Activating Proteins / Q000378:metabolism / Q000502:physiology; D018631:ras Proteins / Q000502:physiology
D018801:CD2 Antigens; D017252:CD3 Complex; D004268:DNA-Binding Proteins; C489560:DOK1 protein, human; C074561:GAP-associated protein p62; D007376:Interleukin-2; D010750:Phosphoproteins; D016601:RNA-Binding Proteins; D020703:ras GTPase-Activating Proteins; D014443:Tyrosine; D018631:ras Proteins; D002118:Calcium