title: Quantitative and temporal requirements revealed for Zap70 catalytic activity during T cell development.
abstract: The catalytic activity of Zap70 is crucial for T cell antigen receptor (TCR) signaling, but the quantitative and temporal requirements for its function in thymocyte development are not known. Using a chemical-genetic system to selectively and reversibly inhibit Zap70 catalytic activity in a model of synchronized thymic selection, we showed that CD4(+)CD8(+) thymocytes integrate multiple, transient, Zap70-dependent signals over more than 36 h to reach a cumulative threshold for positive selection, whereas 1 h of signaling was sufficient for negative selection. Titration of Zap70 activity resulted in graded reductions in positive and negative selection but did not decrease the cumulative TCR signals integrated by positively selected OT-I cells, which revealed heterogeneity, even among CD4(+)CD8(+) thymocytes expressing identical TCRs undergoing positive selection.
authors.affiliation: 1] Howard Hughes Medical Institute, University of California, San Francisco, San Francisco, California, USA. [2] Rosalind Russell-Ephraim P. Engleman Rheumatology Research Center, University of California, San Francisco, San Francisco, California, USA. [3] Department of Medicine, University of California, San Francisco, San Francisco, California, USA. [4] Department of Microbiology and Immunology, University of California, San Francisco, San Francisco, California, USA. [5].
authors.forename: Byron B
authors.identifier: 0000000264469102
authors.initials: BB
authors.lastname: Au-Yeung
authors.affiliation: 1] Division of Immunology and Pathogenesis, Department of Molecular and Cell Biology, University of California Berkeley, Berkeley, California, USA. [2] [3].
authors.forename: Heather J
authors.initials: HJ
authors.lastname: Melichar
authors.affiliation: Division of Immunology and Pathogenesis, Department of Molecular and Cell Biology, University of California Berkeley, Berkeley, California, USA.
authors.forename: Jenny O
authors.initials: JO
authors.lastname: Ross
authors.affiliation: 1] Howard Hughes Medical Institute, University of California, San Francisco, San Francisco, California, USA. [2] Rosalind Russell-Ephraim P. Engleman Rheumatology Research Center, University of California, San Francisco, San Francisco, California, USA. [3] Department of Medicine, University of California, San Francisco, San Francisco, California, USA. [4] Department of Microbiology and Immunology, University of California, San Francisco, San Francisco, California, USA.
authors.forename: Debra A
authors.initials: DA
authors.lastname: Cheng
authors.affiliation: 1] Howard Hughes Medical Institute, University of California, San Francisco, San Francisco, California, USA. [2] Rosalind Russell-Ephraim P. Engleman Rheumatology Research Center, University of California, San Francisco, San Francisco, California, USA. [3] Department of Medicine, University of California, San Francisco, San Francisco, California, USA. [4] Department of Microbiology and Immunology, University of California, San Francisco, San Francisco, California, USA.
authors.forename: Julie
authors.initials: J
authors.lastname: Zikherman
authors.affiliation: 1] Howard Hughes Medical Institute, University of California, San Francisco, San Francisco, California, USA. [2] Department of Cellular and Molecular Pharmacology, University of California, San Francisco, San Francisco, California, USA.
authors.forename: Kevan M
authors.initials: KM
authors.lastname: Shokat
authors.affiliation: Division of Immunology and Pathogenesis, Department of Molecular and Cell Biology, University of California Berkeley, Berkeley, California, USA.
authors.forename: Ellen A
authors.initials: EA
authors.lastname: Robey
authors.affiliation: 1] Howard Hughes Medical Institute, University of California, San Francisco, San Francisco, California, USA. [2] Rosalind Russell-Ephraim P. Engleman Rheumatology Research Center, University of California, San Francisco, San Francisco, California, USA. [3] Department of Medicine, University of California, San Francisco, San Francisco, California, USA. [4] Department of Microbiology and Immunology, University of California, San Francisco, San Francisco, California, USA.
authors.forename: Arthur
authors.initials: A
authors.lastname: Weiss
mesh_terms: D000818:Animals; D002118:Calcium / Q000378:metabolism; D002384:Catalysis; D002454:Cell Differentiation; D047908:Intracellular Signaling Peptides and Proteins / Q000502:physiology; D051379:Mice; D008810:Mice, Inbred C57BL; D011505:Protein-Tyrosine Kinases / Q000502:physiology; D011948:Receptors, Antigen, T-Cell / Q000502:physiology; D015398:Signal Transduction; D000072377:Syk Kinase; D013601:T-Lymphocytes / Q000502:physiology*; D051746:ZAP-70 Protein-Tyrosine Kinase / Q000502:physiology*
chemical_list: D047908:Intracellular Signaling Peptides and Proteins; D011948:Receptors, Antigen, T-Cell; D011505:Protein-Tyrosine Kinases; D000072377:Syk Kinase; C000606897:Syk protein, mouse; D051746:ZAP-70 Protein-Tyrosine Kinase; C497262:Zap70 protein, mouse; D002118:Calcium