Apoptosis but not other activation events is inhibited by a mutation in the transmembrane domain of T cell receptor beta that impairs CD3zeta association.
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 gammadelta 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.
Rodríguez-Tarduchy
G
G
Centro de Biología Molecular Severo Ochoa, CSIC-Universidad Autónoma de Madrid, Cantoblanco, 28049 Madrid, Spain.
Sahuquillo
A G
AG
Alarcón
B
B
Bragado
R
R
D000595:Amino Acid Sequence; D015703:Antigens, CD / Q000378:metabolism; D000945:Antigens, Differentiation, T-Lymphocyte / Q000378:metabolism; D017209:Apoptosis; D001692:Biological Transport; D017252:CD3 Complex / Q000737:chemistry* / Q000502:physiology; D002451:Cell Compartmentation; D002462:Cell Membrane / Q000378:metabolism; D002470:Cell Survival; D016384:Consensus Sequence; D015536:Down-Regulation; D004721:Endoplasmic Reticulum / Q000378:metabolism; D053222:Fas Ligand Protein; D006801:Humans; D007376:Interleukin-2 / Q000378:metabolism; D037181:Lectins, C-Type; D008213:Lymphocyte Activation; D046911:Macromolecular Substances; D008562:Membrane Glycoproteins / Q000378:metabolism; D008969:Molecular Sequence Data; D016693:Receptors, Antigen, T-Cell, alpha-beta / Q000737:chemistry* / Q000502:physiology; D015375:Receptors, Interleukin-2 / Q000378:metabolism; D016415:Sequence Alignment; D015398:Signal Transduction; D013329:Structure-Activity Relationship; D000099192:CD69 Antigens
D015703:Antigens, CD; D000945:Antigens, Differentiation, T-Lymphocyte; D017252:CD3 Complex; D053222:Fas Ligand Protein; D007376:Interleukin-2; D037181:Lectins, C-Type; D046911:Macromolecular Substances; D008562:Membrane Glycoproteins; D016693:Receptors, Antigen, T-Cell, alpha-beta; D015375:Receptors, Interleukin-2; D000099192:CD69 Antigens; C505661:FASLG protein, human