International Immunology, Vol. 13, No. 2, 211-222,
February 2001
© 2001 Japanese Society for Immunology
Conserved transmembrane tyrosine residues of the TCR ß chain are required for TCR expression and function in primary T cells and hybridomas
Department of Immunology, Jerome H. Holland Laboratory for the Biomedical Sciences and George Washington University School of Medicine, American Red Cross, 15601 Crabbs Branch Way, Rockville, MD 20855, USA
1 Wistar Institute, 3601 Spruce Street, Philadelphia, PA 19104, USA
Correspondence to: Correspondence to: L. M. Spain
The T cell receptor (TCR) ß chain transmembrane domain contains two evolutionarily conserved tyrosines (Y). In this study, the functional basis for the evolutionary conservation is addressed by mutation of the residues, expression of the mutants in hybridoma and primary T cells, and examination of TCR signaling function. We find that the phenotype of the mutants, both surface expression and ability to signal for IL-2 production, is highly variable in different mouse T hybridoma lines. Although we have not been able to determine the basis for these differences in the hybridomas, expression of the mutants in primary T cells provides a definitive assessment of mutant phenotype. We show that mutation of the N-terminal Y to either leucine (L) or alanine (A) results in low surface expression in primary T cells, while mutation of both N- and C-terminal Y to A or L abrogates surface expression. However, the more conservative mutation of both transmembrane Y to phenylalanine maintained receptor surface expression and assembly while severely disrupting signaling in primary T cells. Our data demonstrate that TCR ß chain transmembrane Y are essential for TCR signal transduction as well as complex assembly. These findings suggest that protein–protein interactions involving membrane-spanning domains are likely relevant for TCR signal transduction mechanisms.
Keywords: assembly, mutagenesis, signaling, TCR, transmembrane domain
2 Present address: Department of Biology, Cabrini College, 610 King of Prussia Road, Radnor, PA 19087-3698, USA
Transmitting editor: C. Terhorst
CiteULike 
Connotea 
Del.icio.us What's this?
This article has been cited by other articles:
|
|
 |
|
  H. Xu, M. S. Williams, and L. M. Spain Patterns of expression, membrane localization, and effects of ectopic expression suggest a function for MS4a4B, a CD20 homolog in Th1 T cells Blood, March 15, 2006; 107(6): 2400 - 2408. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 T. R. Petersen, S. Gulland, E. Bettelli, V. Kuchroo, E. Palmer, and B. T. Backstrom A chimeric T cell receptor with super-signaling properties Int. Immunol., July 1, 2004; 16(7): 889 - 894. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 E. Teixeiro, P. Fuentes, B. Galocha, B. Alarcon, and R. Bragado T Cell Receptor-mediated Signal Transduction Controlled by the beta Chain Transmembrane Domain. APOPTOSIS-DEFICIENT CELLS DISPLAY UNBALANCED MITOGEN-ACTIVATED PROTEIN KINASES ACTIVITIES UPON T CELL RECEPTOR ENGAGEMENT J. Biol. Chem., February 1, 2002; 277(6): 3993 - 4002. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 L. M. Spain and P. Liu TCR{beta} Transmembrane Tyrosines Are Required for Pre-TCR Function J. Immunol., January 1, 2002; 168(1): 127 - 133. [Abstract] [Full Text] [PDF]
|
|