Sequence and expression of transcripts of the T-cell antigen receptor alpha-chain gene in a functional, antigen-specific suppressor-T-cell hybridoma.
We have detected rearrangement and expression of a gene encoding a T-cell antigen receptor alpha chain in the keyhole limpet hemocyanin (KLH)-specific, inducible suppressor-T-cell (Ts) hybridoma 34S-281 (BW5147 lymphoma-C57BL/6Ts) by using alpha-chain cDNA clones isolated from this Ts hybridoma. The cDNA clones have a restriction length polymorphism in the constant region that identifies them as being of C57BL/6 origin. The cDNA sequence has an ATG start codon for an open reading frame including variable, joining, and constant gene segments. Furthermore, the Ts alpha-chain gene transcripts were detected on membrane-bound polysomes by RNA blot analysis using a variable-region fragment from one of the alpha-chain cDNA clones as probe, suggesting that they are actively translated in the Ts hybridoma. As the beta-chain gene is deleted in all Ts hybridomas analyzed, and since a disulfide-linked dimer is detectable by two-dimensional NaDodSO4/PAGE of lysates of surface-radioiodinated Ts, we suggest that Ts antigen receptors are either alpha-chain homodimers or heterodimers composed of the alpha chain in association with an undefined chain.
D000818:Animals; D001483:Base Sequence; D003001:Cloning, Molecular; D004247:DNA / Q000032:analysis; D004591:Electrophoresis, Polyacrylamide Gel; D006433:Hemocyanins / Q000276:immunology; D006825:Hybridomas / Q000378:metabolism*; D051379:Mice; D008810:Mice, Inbred C57BL; D011948:Receptors, Antigen, T-Cell / Q000096:biosynthesis / Q000235:genetics*; D011995:Recombination, Genetic; D050378:T-Lymphocytes, Regulatory / Q000378:metabolism*; D014158:Transcription, Genetic
D011948:Receptors, Antigen, T-Cell; D004247:DNA; D006433:Hemocyanins; C032808:keyhole-limpet hemocyanin