Skip to main content
Proceedings of the National Academy of Sciences of the United States of America logoLink to Proceedings of the National Academy of Sciences of the United States of America
. 1986 Nov;83(22):8708–8712. doi: 10.1073/pnas.83.22.8708

Sequence and expression of transcripts of the T-cell antigen receptor alpha-chain gene in a functional, antigen-specific suppressor-T-cell hybridoma.

K Imai, M Kanno, H Kimoto, K Shigemoto, S Yamamoto, M Taniguchi
PMCID: PMC387000  PMID: 2946043

Abstract

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.

Full text

PDF
8710

Images in this article

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Arden B., Klotz J. L., Siu G., Hood L. E. Diversity and structure of genes of the alpha family of mouse T-cell antigen receptor. 1985 Aug 29-Sep 4Nature. 316(6031):783–787. doi: 10.1038/316783a0. [DOI] [PubMed] [Google Scholar]
  2. Ballinari D., Castelli C., Traversari C., Pierotti M. A., Parmiani G., Palmieri G., Ricciardi-Castagnoli P., Adorini L. Disulfide-linked surface molecules of monoclonal antigen-specific suppressor T cells: evidence for T cell receptor structures. Eur J Immunol. 1985 Aug;15(8):855–860. doi: 10.1002/eji.1830150822. [DOI] [PubMed] [Google Scholar]
  3. Becker D. M., Pattern P., Chien Y., Yokota T., Eshhar Z., Giedlin M., Gascoigne N. R., Goodnow C., Wolf R., Arai K. Variability and repertoire size of T-cell receptor V alpha gene segments. Nature. 1985 Oct 3;317(6036):430–434. doi: 10.1038/317430a0. [DOI] [PubMed] [Google Scholar]
  4. Benton W. D., Davis R. W. Screening lambdagt recombinant clones by hybridization to single plaques in situ. Science. 1977 Apr 8;196(4286):180–182. doi: 10.1126/science.322279. [DOI] [PubMed] [Google Scholar]
  5. Chien Y., Becker D. M., Lindsten T., Okamura M., Cohen D. I., Davis M. M. A third type of murine T-cell receptor gene. Nature. 1984 Nov 1;312(5989):31–35. doi: 10.1038/312031a0. [DOI] [PubMed] [Google Scholar]
  6. De Santis R., Givol D., Hsu P. L., Adorini L., Doria G., Appella E. Rearrangement and expression of the alpha- and beta-chain genes of the T-cell antigen receptor in functional murine suppressor T-cell clones. Proc Natl Acad Sci U S A. 1985 Dec;82(24):8638–8642. doi: 10.1073/pnas.82.24.8638. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Gershon R. K., Kondo K. Infectious immunological tolerance. Immunology. 1971 Dec;21(6):903–914. [PMC free article] [PubMed] [Google Scholar]
  8. Glisin V., Crkvenjakov R., Byus C. Ribonucleic acid isolated by cesium chloride centrifugation. Biochemistry. 1974 Jun 4;13(12):2633–2637. doi: 10.1021/bi00709a025. [DOI] [PubMed] [Google Scholar]
  9. Goding J. W., Harris A. W. Subunit structure of cell surface proteins: disulfide bonding in antigen receptors, Ly-2/3 antigens, and transferrin receptors of murine T and B lymphocytes. Proc Natl Acad Sci U S A. 1981 Jul;78(7):4530–4534. doi: 10.1073/pnas.78.7.4530. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Goldberg D. A. Isolation and partial characterization of the Drosophila alcohol dehydrogenase gene. Proc Natl Acad Sci U S A. 1980 Oct;77(10):5794–5798. doi: 10.1073/pnas.77.10.5794. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Hayday A. C., Saito H., Gillies S. D., Kranz D. M., Tanigawa G., Eisen H. N., Tonegawa S. Structure, organization, and somatic rearrangement of T cell gamma genes. Cell. 1985 Feb;40(2):259–269. doi: 10.1016/0092-8674(85)90140-0. [DOI] [PubMed] [Google Scholar]
  12. Hedrick S. M., Germain R. N., Bevan M. J., Dorf M., Engel I., Fink P., Gascoigne N., Heber-Katz E., Kapp J., Kaufmann Y. Rearrangement and transcription of a T-cell receptor beta-chain gene in different T-cell subsets. Proc Natl Acad Sci U S A. 1985 Jan;82(2):531–535. doi: 10.1073/pnas.82.2.531. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Hedrick S. M., Nielsen E. A., Kavaler J., Cohen D. I., Davis M. M. Sequence relationships between putative T-cell receptor polypeptides and immunoglobulins. Nature. 1984 Mar 8;308(5955):153–158. doi: 10.1038/308153a0. [DOI] [PubMed] [Google Scholar]
  14. Kronenberg M., Goverman J., Haars R., Malissen M., Kraig E., Phillips L., Delovitch T., Suciu-Foca N., Hood L. Rearrangement and transcription of the beta-chain genes of the T-cell antigen receptor in different types of murine lymphocytes. Nature. 1985 Feb 21;313(6004):647–653. doi: 10.1038/313647a0. [DOI] [PubMed] [Google Scholar]
  15. Marchalonis J. J., Cone R. E., Santer V. Enzymic iodination. A probe for accessible surface proteins of normal and neoplastic lymphocytes. Biochem J. 1971 Oct;124(5):921–927. doi: 10.1042/bj1240921. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Messing J. New M13 vectors for cloning. Methods Enzymol. 1983;101:20–78. doi: 10.1016/0076-6879(83)01005-8. [DOI] [PubMed] [Google Scholar]
  17. Mori L., Lecoq A. F., Robbiati F., Barbanti E., Righi M., Sinigaglia F., Clementi F., Ricciardi-Castagnoli P. Rearrangement and expression of the antigen receptor alpha, beta and gamma genes in suppressor antigen-specific T cell lines. EMBO J. 1985 Aug;4(8):2025–2030. doi: 10.1002/j.1460-2075.1985.tb03887.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Palmiter R. D. Magnesium precipitation of ribonucleoprotein complexes. Expedient techniques for the isolation of undergraded polysomes and messenger ribonucleic acid. Biochemistry. 1974 Aug 13;13(17):3606–3615. doi: 10.1021/bi00714a032. [DOI] [PubMed] [Google Scholar]
  19. Ramsey J. C., Steele W. J. Differences in size, structure and function of free and membrane-bound polyribosomes of rat liver. Evidence for a single class of membrane-bound polyribosomes. Biochem J. 1977 Oct 15;168(1):1–8. doi: 10.1042/bj1680001. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Saito H., Kranz D. M., Takagaki Y., Hayday A. C., Eisen H. N., Tonegawa S. A third rearranged and expressed gene in a clone of cytotoxic T lymphocytes. Nature. 1984 Nov 1;312(5989):36–40. doi: 10.1038/312036a0. [DOI] [PubMed] [Google Scholar]
  21. Saito H., Kranz D. M., Takagaki Y., Hayday A. C., Eisen H. N., Tonegawa S. Complete primary structure of a heterodimeric T-cell receptor deduced from cDNA sequences. 1984 Jun 28-Jul 4Nature. 309(5971):757–762. doi: 10.1038/309757a0. [DOI] [PubMed] [Google Scholar]
  22. Southern E. M. Detection of specific sequences among DNA fragments separated by gel electrophoresis. J Mol Biol. 1975 Nov 5;98(3):503–517. doi: 10.1016/s0022-2836(75)80083-0. [DOI] [PubMed] [Google Scholar]
  23. Sumida T., Takei I., Taniguchi M. Activation of acceptor-suppressor hybridoma with antigen-specific suppressor T cell factor of two-chain type: requirement of the antigen- and the I-J-restricting specificity. J Immunol. 1984 Sep;133(3):1131–1136. [PubMed] [Google Scholar]
  24. Sumida T., Taniguchi M. Novel mechanisms of specific suppression of anti-hapten antibody response mediated by monoclonal anti-carrier antibody. J Immunol. 1985 Jun;134(6):3675–3681. [PubMed] [Google Scholar]
  25. Tada T., Okumura K. The role of antigen-specific T cell factors in the immune response. Adv Immunol. 1979;28:1–87. doi: 10.1016/s0065-2776(08)60799-3. [DOI] [PubMed] [Google Scholar]
  26. Taniguchi M., Sumida T. "I-J" as an idiotypic marker on the antigen-specific suppressor T cell factor. Immunol Rev. 1985 Apr;83:125–150. doi: 10.1111/j.1600-065x.1985.tb00473.x. [DOI] [PubMed] [Google Scholar]
  27. Yanagi Y., Yoshikai Y., Leggett K., Clark S. P., Aleksander I., Mak T. W. A human T cell-specific cDNA clone encodes a protein having extensive homology to immunoglobulin chains. Nature. 1984 Mar 8;308(5955):145–149. doi: 10.1038/308145a0. [DOI] [PubMed] [Google Scholar]

Articles from Proceedings of the National Academy of Sciences of the United States of America are provided here courtesy of National Academy of Sciences

RESOURCES