Molecular mechanisms in the TCR (TCR alpha beta-CD3 delta epsilon, gamma epsilon) interaction with zeta 2 homodimers: clues from a 'phenotypic revertant' clone.
The association between the TCRalphabeta-CD3gammaepsilondeltaepsilon hexamers and zeta2 homodimers in the endoplasmic reticulum (ER) constitutes a key step in TCR assembly and export to the T cell surface. Incompletely assembled TCR-CD3 complexes are degraded in the ER or the lysosomes. A previously described Jurkat variant (J79) has a mutation at position 195 on the TCR Calpha domain causing a phenylalanine to valine exchange. This results in a lack of association between TCRalphabeta-CD3gammaepsilondeltaepsilon hexamers and zeta2 homodimers. Two main hypotheses could explain this phenomenon in J79 cells: TCR-CD3 hexamers may be incapable of interacting with zeta2 due to a structural change in the TCR Calpha region; alternatively, TCR-CD3 hexamers may be incapable of interacting with zeta2 due to factors unrelated to either molecular complex. In order to assess these two possibilities, the TCR-CD3 membrane-negative J79 cells were treated with ethylmethylsulfonate and clones positive for TCR membrane expression were isolated. The characterization of the J79r58 phenotypic revertant cell line is the subject of this study. The main question was to assess the reason for the TCR re-expression. The TCR on J79r58 cells appears qualitatively and functionally equivalent to wild-type TCR complexes. Nucleotide sequence analysis confirmed the presence of the original mutation in the TCR Calpha region but failed to detect compensatory mutations in alpha, beta, gamma, delta, epsilon or zeta chains. Thus, mutated J79-TCR-CD3 complexes can interact with zeta2 homodimers. Possible mechanisms for the unsuccessful TCR-CD3 interaction with zeta2 homodimers are presented and discussed.
Martin
E P
EP
Unité de Physiopathologie Cellulaire et Moléculaire, CNRS, ERS 1590, IFR 30 d'Immunologie Cellulaire et Moléculaire, CHU de Purpan, 31059 cedex 03 Toulouse, France.
Arnaud
J
J
Alibaud
L
L
Gouaillard
C
C
Llobera
R
R
Huchenq-Champagne
A
A
Rubin
B
B
D002462:Cell Membrane / Q000378:metabolism; D002999:Clone Cells; D019281:Dimerization; D005434:Flow Cytometry; D006801:Humans; D019169:Jurkat Cells; D046911:Macromolecular Substances; D018832:Molecular Chaperones / Q000378:metabolism / Q000502:physiology; D009154:Mutation; D010641:Phenotype; D017260:Receptor-CD3 Complex, Antigen, T-Cell / Q000096:biosynthesis / Q000235:genetics / Q000378:metabolism* / Q000502:physiology; D016693:Receptors, Antigen, T-Cell, alpha-beta / Q000378:metabolism* / Q000502:physiology; D015398:Signal Transduction / Q000276:immunology
D046911:Macromolecular Substances; D018832:Molecular Chaperones; D017260:Receptor-CD3 Complex, Antigen, T-Cell; D016693:Receptors, Antigen, T-Cell, alpha-beta