Quantitative and temporal requirements revealed for Zap70 catalytic activity during T cell development

Byron B Au-Yeung; Heather J Melichar; Jenny O Ross; Debra A Cheng; Julie Zikherman; Kevan M Shokat; Ellen A Robey; Arthur Weiss

doi:10.1038/ni.2918

Quantitative and temporal requirements revealed for Zap70 catalytic activity during T cell development

Nat Immunol. 2014 Jul;15(7):687-94. doi: 10.1038/ni.2918. Epub 2014 Jun 8.

Authors

Byron B Au-Yeung¹, Heather J Melichar², Jenny O Ross³, Debra A Cheng⁴, Julie Zikherman⁴, Kevan M Shokat⁵, Ellen A Robey³, Arthur Weiss⁴

Affiliations

¹ 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].
² 1] Division of Immunology and Pathogenesis, Department of Molecular and Cell Biology, University of California Berkeley, Berkeley, California, USA. [2] [3].
³ Division of Immunology and Pathogenesis, Department of Molecular and Cell Biology, University of California Berkeley, Berkeley, California, USA.
⁴ 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 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.

PMID: 24908390
PMCID: PMC4095875
DOI: 10.1038/ni.2918

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.

Publication types

Research Support, N.I.H., Extramural
Research Support, Non-U.S. Gov't

MeSH terms

Animals
Calcium / metabolism
Catalysis
Cell Differentiation
Intracellular Signaling Peptides and Proteins / physiology
Mice
Mice, Inbred C57BL
Protein-Tyrosine Kinases / physiology
Receptors, Antigen, T-Cell / physiology
Signal Transduction
Syk Kinase
T-Lymphocytes / physiology*
ZAP-70 Protein-Tyrosine Kinase / physiology*

Substances

Intracellular Signaling Peptides and Proteins
Receptors, Antigen, T-Cell
Protein-Tyrosine Kinases
Syk Kinase
Syk protein, mouse
ZAP-70 Protein-Tyrosine Kinase
Zap70 protein, mouse
Calcium

Abstract

Publication types

MeSH terms

Substances

Grants and funding