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Volume 271, Number 48, Issue of November 29, 1996 pp. 30417-30425
©1996 by The American Society for Biochemistry and Molecular Biology, Inc.

Apoptosis but Not Other Activation Events Is Inhibited by a Mutation in the Transmembrane Domain of T Cell Receptor beta  That Impairs CD3zeta Association

(Received for publication, February 27, 1996, and in revised form, August 12, 1996)

Gemma Rodríguez-Tarduchy Dagger , Almudena G. Sahuquillo , Balbino Alarcón Dagger and Rafael Bragado

From the Dagger  Centro de Biología Molecular Severo Ochoa, CSIC-Universidad Autónoma de Madrid, Cantoblanco, 28049 Madrid and the  Department of Immunology, Fundación Jiménez-Díaz, Avenida Reyes Católicos 2, 28040 Madrid, Spain

The transmembrane domain of T cell receptor (TCR) beta  contains a conserved immunoreceptor tyrosine-based activation-like motif consisting of a duplicated YXXL sequence. The motif is also present in TCRgamma , the equivalent chain to TCRbeta in gamma delta T lymphocytes but is absent in TCRalpha and TCRdelta . To determine the putative role of this sequence in TCR·CD3 complex assembly and signal transduction, a TCRbeta chain cDNA was mutated in the C-terminal tyrosine of the motif, cloned in an expression vector, and transfected into TCRbeta -negative Jurkat cells. Transfectants of the mutated chain (MUT) expressed, on average, much less TCR·CD3 complex on the membrane than wild type TCRbeta transfectants. Radiolabeling experiments suggested that the mutation caused a loose association of the CD3zeta chain resulting in a defective assembly. However, stimulation of high TCR·CD3 expressing wild type and MUT clones with monoclonal antibodies and Staphylococcus aureus enterotoxin B resulted in similar levels of CD25 and CD69 expression, interleukin-2 secretion, and TCR·CD3 complex down-regulation. By contrast, MUT cells were clearly resistant to activation-induced cell death, and they did not express CD95-ligand upon activation. These results suggest a differentiated intracellular signaling pathway leading to apoptosis in which Tyr-TM11 of the immunoreceptor tyrosine-based activation motif-like motif and CD3zeta appear to be involved.


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