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Structure and expression of the Lyt-3 agene of C.AKR mice

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Abstract

The mouse Lyt-3 agene, which encodes the Lyt-3.1 T-cell surface alloantigen of the C.AKR strain, has been cloned, and the nucleotide sequence of its exons and more than 2 kb of 5′ flanking sequence have been determined. The gene extends over approximately 16 kb of DNA and consists of six exons encoding leader, leader plus V-like domain, membrane-proximal, transmembrane, and cytoplasmic domains. The only difference between the coding region of the Lyt-3 agene and the cDNA sequences reported for Lyt-3 b(Nakauchi et al. 1987, Panaccio et al. 1987) is at position 77 of the mature protein where Lyt-3 aencodes serine and Lyt-3 bencodes arginine. This substitution must therefore be the basis for the serological distinction between the Lyt-3.1 and Lyt-3.2 alloantigens. Potential TATA and CAAT sequences, two Sp1 protein binding sites, two extended repeats of the dinucleotide, CA, a number of short inverted repeats, and an inverted segment of the mouse B1 repetitive sequence are found 5′ to the Lyt-3 agene. Two consensus poly-A addition signals and a complete copy of the mouse B1 sequence are found 3′ to the gene. Both B1-related regions are flanked by short direct repeats suggesting that they arose by an insertional mechanism. Cotransfection of the Lyt-3 agene together with a cloned Lyt-2 agene resulted in expression of both Lyt-2 and Lyt-3.1 on the surface of Ltk and BW5147 cells. Transfection of the Lyt-3 agene without Lyt-2 aled to expression of Lyt-3-related cellular RNA but did not result in surface expression of Lyt-3.1, suggesting that the Lyt-3 glycoprotein is not expressed on the cell surface in the absence of Lyt-2.

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References

  • Boyd, R. R., Goldrick, M. M., and Gottlieb, P. D.: Genetic polymorphism at the mouse immunoglobulin J κ locus (Igk-J) as demonstrated by Southern hybridization and nucleotide sequence analysis. Immunogenetics 24: 150–157, 1986a

    Google Scholar 

  • Boyd, R. T., Goldrick, M. M., and Gottlieb, P. D.: Structural differences in a single gene encoding the VκSer group of light chains explain the existence of two mouse light-chain genetic markers. Proc Natl Acad Sci USA 83: 9134–9138, 1986b

    Google Scholar 

  • Boyse, E. A., Miyazawa, M., Aoki, T., and Old, L. J.: Ly-A and Ly-B: two systems of lymphocyte isoantigens in the mouse. Proc R Soc London 170: 175–193, 1968

    Google Scholar 

  • Boyse, E. A., Itakura, K., Stockert, E., Iritani, C. A., and Miura, M.: LyC: a third locus specifying alloantigens exposed only on thymocytes and lymphocytes. Transplantation 11: 351–352, 1971

    Google Scholar 

  • Dembic, Z., Haas, W., Zamoyska, R., Parties, J., Steinmetz, M., and von Boehmer, H.: Transfection of the CD8 gene enhances T-cell recognition. Nature 326: 510–511, 1987

    Google Scholar 

  • Durda, P. J. and Gottlieb, P. D.: The Ly-3 antigens on mouse thymocytes: immune precipitation and molecular weight characterization. J Exp Med 144: 476–493, 1976

    Google Scholar 

  • Durda, P. J. and Gottlieb, P. D.: Sequential precipitation of mouse thymocyte extracts with anti-Lyt-2 and anti-Lyt-3 sera. I. Lyt-2.1 and Lyt-3.1 antigenic determinants reside on separable molecular species. J Immunol 121: 983–989, 1978

    Google Scholar 

  • Frischauf, A.-M., Lehrach, H., Poustka, A., and Murray, N.: Lambda replacement vectors carrying polylinker sequences. J Mol Biol 170: 827–842, 1983

    Google Scholar 

  • Gabert, J., Langlet, C., Zamoyska, R., Parnes, J. R., Schmitt-Verhulst, A.-M., and Malissen, B.: Reconstitution of the MHC class I specificity by transfer of the T cell receptor and Lyt-2 genes. Cell 50: 545–554, 1987

    Google Scholar 

  • Gillies, S. D., Morrison, S. L., Oi, V. T., and Tonegawa, S.: A tissue-specific transcription enhancer element is located in the major intron of a rearranged immunoglobulin heavy chain gene. Cell 33: 717–728, 1983

    Google Scholar 

  • Gottlieb, P. D.: Genetic correlation of a mouse light chain variable region marker with a thymocyte surface antigen. J Exp Med 140: 1432–1437, 1974

    Google Scholar 

  • Gottlieb, P. D., Marshak-Rothstein, A., Auditore-Hargreaves, K., Berkoben, D. B., August, D. A., Rosche R. M., and Benedetto, J. D.: Construction and properties of new Lyt-congenic strains and anti-Lyt-2.2 and anti-Lyt-3.1 monoclonal antibodies. Immunogenetics 10: 545–555, 1980

    Google Scholar 

  • Graham, F. L., Bacchetti, S., McKinnon, R., Stanners, C., Cordell, B., and Goodman, H. M.: Transformation of mammaliam cells with DNA using the calcium technique. In R. Baserga, C. Croce, and G. Rovera (eds.) Introduction of Macromolecules into Viable Mammaliam Cells, pp. 3–25, Alan R. Liss, Inc., New York, 1980

    Google Scholar 

  • Itakura, K., Hutton, J. J., Boyse, E. A., and Old, L. J.,: Genetic linkage relationships of loci specifying differentiation alloantigens in the mouse. Transplantation 13: 239–243, 1972

    Google Scholar 

  • IUPAC-IUB Commission on Biochemical Nomenclature: A one letter notation for amino acid sequences: tentative rules. J Biol Chem 243: 3557–3559, 1968

    Google Scholar 

  • Johnson, P.: A human homolog of the mouse CD8 molecule, Lyt-3: genomic sequence and expression. Immunogenetics 26: 174–177, 1987

    Google Scholar 

  • Johnson, P. and Williams, A. F.: Striking similarities between antigen receptor J pieces and sequence in the second chain of the muringe CD8 antigen. Nature 323: 74–76, 1986

    Google Scholar 

  • Johnson, P., Gagnon, L., Barclay, A. N., and Williams, A. F.: Purification, chain separation and sequence of the MRC OX-8 antigen, a marker of rat cytotoxic T lymphocytes. EMBO J 4: 2539–2545, 1985

    Google Scholar 

  • Kadonaga, J. T., Jones, K. A., and Tijan, R.: Promoter-specific activation of RNA polymerase II transcription by Spl. Trends Biochem 11: 20–23, 1986

    Google Scholar 

  • Kavathas, P. and Herzenberg, L. A.: Transfection for lymphocyte cell surface antigens. In D. M. Weir (ed.): Handbook of Experimental Immunology, Fourth Edition, Volume 3, Blackwell Scientific Publications, Oxford, pp. 91.1–91.10, 1986

    Google Scholar 

  • Kavathas, P., Sukhatme, V. P., Herzenberg, L. A., and Parties, J. R.: Isolation of a gene encoding the human T-lymphocyte differentiation antigen Leu-2 (T8) by gene transfer and cDNA subtraction. Proc Natl Acad Sci USA 81: 7688–7692, 1984

    Google Scholar 

  • Krayev, A. S., Kramerov, D. A., Skryabin, K. G., Ryskov, A. P., Bayev, A. A., and Georgiev, G. P.: The nucleotide sequence of the ubiquitous repetetive DNA sequence B 1 complementary to the most abundant class of mouse fold-back RNA. Nucleic Acids Res 8: 1201–1215, 1980

    Google Scholar 

  • Ledbetter, J. A. and Herzenberg, L. A.: Xenogeneic monoclonal antibodies to mouse lymphoid differentiation antigens. Immunol Rev 47: 63–90, 1979

    Google Scholar 

  • Ledbetter, J. A., Seaman, W. E., Tsu, T. T., and Herzenberg, L. A.: Lyt-2 and Lyt-3 antigens are on two different polypeptide subunits linked by disulfide bonds. Relationship of subunits to T cell cytolytic activity. J Exp Med 153: 1503–1516, 1981

    Google Scholar 

  • Liaw, C. W., Zamoyska, R., and Parties, P. R.: Structure, sequence, and polymorphism of the Lyt-2 T cell differentiation gene. J Immunol 137: 1037–1043, 1986

    Google Scholar 

  • MacDonald, H. R., Glasebrook, A. L., Bron, C., Kelso, A., and Cerottini, J.-C.: Clonal heterogeneity in the functional requirement for Lyt-2/3 molecules on cytolytic T lymphocytes (CTL): possible implications for the affinity of CTL antigen receptors. Immunol Rev 68: 88–115, 1982

    Google Scholar 

  • Maniatis, T., Fritsch, E. F., and Sambrook, J.: Molecular Cloning: A Laboratory Manual, pp. 1–545, Cold Spring Harbor Laboratory, Cold Spring Harbor, 1983

    Google Scholar 

  • Manser, T. and Getter, M. L.: Isolation of hybridomas expressing a specific heavy chain variable region gene segment by using a screening technique that detects mRNA sequences in whole cell lysates. Proc Natl Acad Sci USA 81: 2470–2474, 1984

    Google Scholar 

  • Messing, J.: M13mp2 and derivatives: a molecular cloning system for DNA sequencing, strand-specific hybridization, and in vitro mutagenesis. In A. Walton (ed.): Third Cleveland Symposium on Macromolecules: Recombinant DNA, pp. 143–153, Elsevier North Holland, Amsterdam, 1981

    Google Scholar 

  • Mulligan, R. C. and Berg, P.: Expression of a bacterial gene in mammalian cells. Science 209: 1422–1427, 1980

    Google Scholar 

  • Nakauchi, H., Shinkai, Y., and Okumura, K.: Molecular cloning of Lyt-3, a membrane glycoprotein marking a subset of mouse T lymphocytes: molecular homology to immunoglobulin and T-cell receptor variable and joining regions. Proc Natl Acad Sci USA 84: 4210–4214, 1987

    Google Scholar 

  • Novak, T. J. and Rothenberg, E. V.: Differential transient and longterm expression of DNA sequences introduced into T lymphocyte lines. DNA 5: 439–451, 1986

    Google Scholar 

  • Panaccio, M., Gillespie, M. T., Walker, I. D., Kirszbaum, L., Sharpe, J. A., Tobias, G. H., McKenzie, I. F. C., and Deacon, N. J.: Molecular characterization of the murine cytotoxic T-cell mem brane glycoprotein Ly-3 (CD8). Proc Natl Acad Sci USA 84: 6874–6878, 1987

    Google Scholar 

  • Poncz, M., Solowiejczyk, D., Ballantine, M., Schwartz, E., and Surrey, S.: “Non-random” DNA sequence analysis in bacteriophage M13 by the dideoxy chain-termination method. Proc Natl Acad Sci USA 79: 4298–4302, 1982

    Google Scholar 

  • Potter, H., Weir, L., and Leder, P.: Enhancer-dependent expression of human immunoglobulin genes introduced into mouse pre-B lymphocytes by electroporation. Proc Natl Acad Sci USA 81: 7161–7165, 1984

    Google Scholar 

  • Reilly, E. B., Auditore-Hargreaves, K., Hämmerling, U., and Gottlieb, P. D.: Lyt-2 and Lyt-3 alloantigens: precipitation with monoclonal and conventional antibodies and analysis on one- and two-dimensional polyacrylamide gels. J Immunol 125: 2245–2251, 1980

    Google Scholar 

  • Rich, A., Nordheim, A., and Wang, A. H.-J.: The chemistry and biology of left-handed Z-DNA. Annu Rev Biochem 53: 791–846, 1984

    Google Scholar 

  • Sandri-Goldin, R. M., Goldin, A. L., Levine, M., and Glorioso, J. C.: High frequency transfer of cloned Herpes Simplex virus type I sequences to mammaliam cells by protoplast fusion. Mol Cell Biol 1: 743–752, 1981

    Google Scholar 

  • Sanger, F., Nicklen, S., and Coulson, A. R.: DNA sequencing with chain-terminating inhibitors. Proc Natl Acad Sci USA 74: 5463–5467, 1977

    Google Scholar 

  • Shimonkevitz, R., Leuscher, B., Cerottini, J.-C., and MacDonald, H. R.: Clonal analysis of cytolytic T lymphocyte-mediated lysis of target cells with inducible antigen expression: correlation between antigen density and requirement for Lyt-2/3 function. J Immunol 135: 892–899, 1985

    Google Scholar 

  • Shinohara, N. and Sachs, D.: Mouse alloantibodies capable of blocking cytotoxic T cell function. I. Relationship between the antigen reactive with blocking antibodies and the Lyt-2 locus. J Exp Med 150: 432–444, 1979

    Google Scholar 

  • Swain, S. L.: Significance of Lyt phenotype: Lyt-2 antibodies block activities of T cells that recognize class I major histocompatibility complex antigens regardless of their function. Proc Natl Acad Sci USA 78: 7101–7105, 1981

    Google Scholar 

  • Swain, S. L.: T cell subsets and the recognition of MHC class. Immunol Rev 74: 129–142, 1983

    Google Scholar 

  • Walker, I. D., Murray, B. J., Hogarth, P. M., Kelso, A., and McKenzie, I. F. C.: Comparison of thymic and peripheral T cell Lyt-2/3 antigens. Eur J Immunol 14: 906–910, 1984

    Google Scholar 

  • Walker, I. D., Murray, B. J., Kirszbaum, L., Chambers, G. W., Deacon, N. J., and McKenzie, I. F. C.: The amino-terminal sequences of Lyt-2 and Lyt-3. Immunogenetics 23: 60–63, 1986

    Google Scholar 

  • Weiher, H., König, M., and Gruss, P.: Multiple point mutations affecting the simian virus 40 enhancer. Science 219: 626–631, 1983

    Google Scholar 

  • Youn, H. J., Harriss, J. V., and Gottlieb, P. D.: Nucleotide sequence analysis of the C.AKR Lyt-2 agene: structural polymorphism in alleles encoding the Lyt-2.1 T-cell surface alloantigen. Immunogenetics, in press, 1988

  • Zamoyska, R., Vollmer, A. C., Sizer, K. C., Liaw C. W., and Parnes, J. R.: Two Lyt-2 polypeptides arise from a single gene by alternative splicing patterns of mRNA. Cell 43: 153–163, 1985

    Google Scholar 

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Youn, H.J., Harriss, J.V. & Gottlieb, P.D. Structure and expression of the Lyt-3 agene of C.AKR mice. Immunogenetics 28, 353–361 (1988). https://doi.org/10.1007/BF00364234

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