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Mol Cell Biol, March 1998, p. 1388-1399, Vol. 18, No. 3
Department of Immunology, Mayo Clinic,
Guggenheim (3), Rochester, Minnesota 559051;
Institut de Cardiologie de Montreal, Montreal, Quebec, Canada
H1T 1C82; and
Cell Biology and
Metabolism Branch, National Institutes of Health, Bethesda, Maryland
20892-54303
Received 16 July 1997/Returned for modification 26 August
1997/Accepted 8 December 1997
T-cell antigen receptor (TCR) engagement activates multiple protein
tyrosine kinases (PTKs), including the Src family member, Lck, and the
Syk-related PTK, ZAP-70. Studies in ZAP-70-deficient humans have
demonstrated that ZAP-70 plays crucial roles in T-cell activation and
development. However, progress toward a detailed understanding of the
regulation and function of ZAP-70 during TCR signaling has been
hampered by the lack of a suitable T-cell model for biochemical and
genetic analyses. In this report, we describe the isolation and
phenotypic characterization of a Syk- and ZAP-70-negative somatic
mutant derived from the Jurkat T-cell line. The P116 cell line displays
severe defects in TCR-induced signaling functions, including protein
tyrosine phosphorylation, intracellular Ca2+ mobilization,
and interleukin-2 promoter-driven transcription. These signaling
defects were fully reversed by reintroduction of catalytically active
versions of either Syk or ZAP-70 into the P116 cells. However, in
contrast to ZAP-70 expression, Syk expression triggered a significant
degree of cellular activation in the absence of TCR ligation.
Transfection experiments with ZAP-70-Syk chimeric proteins indicated
that both the amino-terminal regulatory regions and the
carboxy-terminal catalytic domains of Syk and ZAP-70 contribute to the
distinctive functional properties of these PTKs. These studies
underscore the crucial role of ZAP-70 in TCR signaling and offer a
powerful genetic model for further analyses of ZAP-70 regulation and
function in T cells.
0270-7306/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.
Genetic Evidence for Differential Coupling of Syk
Family Kinases to the T-Cell Receptor: Reconstitution Studies in a
ZAP-70-Deficient Jurkat T-Cell Line
*
Corresponding author. Mailing address: Department of
Immunology, 301 Guggenheim Building, Mayo Clinic, Rochester, MN 55905.
Present address: Gerontology Research Center, National Institute on
Aging, Baltimore, MD 21224.
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