TY - JOUR
T1 - Phosphorylation of multiple CD3ζ tyrosine residues leads to formation of pp21 in vitro and in vivo
T2 - Structural changes upon T cell receptor stimulation
AU - Koyasu, Shigeo
AU - McConkey, David J.
AU - Clayton, Linda K.
AU - Abraham, Sheena
AU - Yandava, Booma
AU - Katagiri, Takuya
AU - Moingeon, Philippe
AU - Yamamoto, Tadashi
AU - Reinherz, Ellis L.
N1 - Copyright:
Copyright 2004 Elsevier B.V., All rights reserved.
PY - 1992/2/15
Y1 - 1992/2/15
N2 - T lymphocyte activation resulting from antigen recognition involves a protein tyrosine kinase pathway which triggers phosphorylation of several cellular substrates including the CD3ζ subunit of the T cell receptor (TCR) to form pp21. The homologous TCR-associated protein, CD3η, is an alternatively spliced product of the same gene locus as CD3ζ. CD3η lacks one of six cytoplasmic tyrosine residues (Tyr-132) found in CD3ζ and is itself not phosphorylated. Site-directed mutagenesis in conjunction with in vitro and in vivo phosphorylation studies herein demonstrates that Tyr-132 is required for the formation of pp21. Moreover, the differential phosphorylation of CD3ζ versus CD3η is not due to a selective association of the known TCR-associated protein tyrosine kinase, p59fyn; p59fyn but not p56lck or p62yes is associated with each of the three TCR isoforms containing CD3ζ2, CD3η2, or CD3ζ-η. This association occurs through components of the TCR complex distinct from CD3ζ or CD3η. In addition, we show that pp21 formation is not only dependent on Tyr-132 but results from concomitant phosphorylation of other CD3ζ residues including Tyr-121. Mutation of Tyr-90, -121, or -132 does not alter primary signal transduction as shown by the ability of individual CD3ζ Tyr → Phe mutants to produce interleukin-2 upon TCR stimulation. Thus, the substantial structural changes in CD3ζ upon TCR stimulation as reflected by alteration in its mobility in sodium dodecyl sulfate-polyacrylamide gel electrophoresis may affect subsequent events such as receptor desensitization, receptor movement, and/or protein associations.
AB - T lymphocyte activation resulting from antigen recognition involves a protein tyrosine kinase pathway which triggers phosphorylation of several cellular substrates including the CD3ζ subunit of the T cell receptor (TCR) to form pp21. The homologous TCR-associated protein, CD3η, is an alternatively spliced product of the same gene locus as CD3ζ. CD3η lacks one of six cytoplasmic tyrosine residues (Tyr-132) found in CD3ζ and is itself not phosphorylated. Site-directed mutagenesis in conjunction with in vitro and in vivo phosphorylation studies herein demonstrates that Tyr-132 is required for the formation of pp21. Moreover, the differential phosphorylation of CD3ζ versus CD3η is not due to a selective association of the known TCR-associated protein tyrosine kinase, p59fyn; p59fyn but not p56lck or p62yes is associated with each of the three TCR isoforms containing CD3ζ2, CD3η2, or CD3ζ-η. This association occurs through components of the TCR complex distinct from CD3ζ or CD3η. In addition, we show that pp21 formation is not only dependent on Tyr-132 but results from concomitant phosphorylation of other CD3ζ residues including Tyr-121. Mutation of Tyr-90, -121, or -132 does not alter primary signal transduction as shown by the ability of individual CD3ζ Tyr → Phe mutants to produce interleukin-2 upon TCR stimulation. Thus, the substantial structural changes in CD3ζ upon TCR stimulation as reflected by alteration in its mobility in sodium dodecyl sulfate-polyacrylamide gel electrophoresis may affect subsequent events such as receptor desensitization, receptor movement, and/or protein associations.
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M3 - Article
C2 - 1531339
AN - SCOPUS:0026744709
SN - 0021-9258
VL - 267
SP - 3375
EP - 3381
JO - Journal of Biological Chemistry
JF - Journal of Biological Chemistry
IS - 5
ER -