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. 1987 May 1;165(5):1358–1370. doi: 10.1084/jem.165.5.1358

Tissue-specific expression of cell-surface Qa-2 antigen from a transfected Q7b gene of C57BL/10 mice

PMCID: PMC2188306  PMID: 3502706

Abstract

We screened a cDNA library prepared from a BALB.B10 CTL clone that expresses Qa-2 antigen, and isolated four clones derived from Q7b, a Qa region gene of C57BL/10. One of these Q7b cDNAs and the Q7b chromosomal gene were subcloned into expression vectors and transfected into L cells and R1.1 thymoma cells. We found that the chromosomal Q7b gene expresses Qa-2 on the surface of R1.1 cells, but not on L cells while the Q7b cDNA expresses protein on the surface of both cell types. The levels of Qa-2 expression do not correlate with the total levels of Q7b mRNA in these transfectants. Our results suggest that the tissue- specific expression of Qa-2 may be controlled, in part, by mechanisms of alternate RNA splicing. By using hybrid gene constructs, we have mapped the tissue-specific element to the 3' part of the gene, downstream of a site near the middle of exon 4. The hybrid polypeptides differ significantly in their transmembrane and cytoplasmic regions. These portions of the protein also may play a role in the tissue- specific expression of Qa-2.

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Selected References

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  1. Allen H., Fraser J., Flyer D., Calvin S., Flavell R. Beta 2-microglobulin is not required for cell surface expression of the murine class I histocompatibility antigen H-2Db or of a truncated H-2Db. Proc Natl Acad Sci U S A. 1986 Oct;83(19):7447–7451. doi: 10.1073/pnas.83.19.7447. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Allen H., Wraith D., Pala P., Askonas B., Flavell R. A. Domain interactions of H-2 class I antigens alter cytotoxic T-cell recognition sites. Nature. 1984 May 17;309(5965):279–281. doi: 10.1038/309279a0. [DOI] [PubMed] [Google Scholar]
  3. Auffray C., Rougeon F. Purification of mouse immunoglobulin heavy-chain messenger RNAs from total myeloma tumor RNA. Eur J Biochem. 1980 Jun;107(2):303–314. doi: 10.1111/j.1432-1033.1980.tb06030.x. [DOI] [PubMed] [Google Scholar]
  4. Cate R. L., Mattaliano R. J., Hession C., Tizard R., Farber N. M., Cheung A., Ninfa E. G., Frey A. Z., Gash D. J., Chow E. P. Isolation of the bovine and human genes for Müllerian inhibiting substance and expression of the human gene in animal cells. Cell. 1986 Jun 6;45(5):685–698. doi: 10.1016/0092-8674(86)90783-x. [DOI] [PubMed] [Google Scholar]
  5. Church G. M., Gilbert W. Genomic sequencing. Proc Natl Acad Sci U S A. 1984 Apr;81(7):1991–1995. doi: 10.1073/pnas.81.7.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Devlin J. J., Weiss E. H., Paulson M., Flavell R. A. Duplicated gene pairs and alleles of class I genes in the Qa2 region of the murine major histocompatibility complex: a comparison. EMBO J. 1985 Dec 1;4(12):3203–3207. doi: 10.1002/j.1460-2075.1985.tb04066.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Flavell R. A., Allen H., Burkly L. C., Sherman D. H., Waneck G. L., Widera G. Molecular biology of the H-2 histocompatibility complex. Science. 1986 Jul 25;233(4762):437–443. doi: 10.1126/science.3726537. [DOI] [PubMed] [Google Scholar]
  8. Goodenow R. S., McMillan M., Nicolson M., Sher B. T., Eakle K., Davidson N., Hood L. Identification of the class I genes of the mouse major histocompatibility complex by DNA-mediated gene transfer. Nature. 1982 Nov 18;300(5889):231–237. doi: 10.1038/300231a0. [DOI] [PubMed] [Google Scholar]
  9. Grosveld F. G., Lund T., Murray E. J., Mellor A. L., Dahl H. H., Flavell R. A. The construction of cosmid libraries which can be used to transform eukaryotic cells. Nucleic Acids Res. 1982 Nov 11;10(21):6715–6732. doi: 10.1093/nar/10.21.6715. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. 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]
  11. Kincade P. W., Flaherty L., Lee G., Watanabe T., Michaelson J. Qa antigen expression on functional lymphoid, myeloid, and stem cells in adult mice. J Immunol. 1980 Jun;124(6):2879–2885. [PubMed] [Google Scholar]
  12. Lemke H., Hämmerling G. J., Hämmerling U. Fine specificity analysis with monoclonal antibodies of antigens controlled by the major histocompatibility complex and by the Qa/TL region in mice. Immunol Rev. 1979;47:175–206. doi: 10.1111/j.1600-065x.1979.tb00293.x. [DOI] [PubMed] [Google Scholar]
  13. Lynes M. A., Tonkonogy S., Flaherty L. Qa-1 and Qa-2 expression on CFU-s. J Immunol. 1982 Sep;129(3):928–930. [PubMed] [Google Scholar]
  14. Maxam A. M., Gilbert W. Sequencing end-labeled DNA with base-specific chemical cleavages. Methods Enzymol. 1980;65(1):499–560. doi: 10.1016/s0076-6879(80)65059-9. [DOI] [PubMed] [Google Scholar]
  15. Mellor A. L., Antoniou J., Robinson P. J. Structure and expression of genes encoding murine Qa-2 class I antigens. Proc Natl Acad Sci U S A. 1985 Sep;82(17):5920–5924. doi: 10.1073/pnas.82.17.5920. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Mellor A. L., Weiss E. H., Kress M., Jay G., Flavell R. A. A nonpolymorphic class I gene in the murine major histocompatibility complex. Cell. 1984 Jan;36(1):139–144. doi: 10.1016/0092-8674(84)90082-5. [DOI] [PubMed] [Google Scholar]
  17. Ozato K., Sachs D. H. Monoclonal antibodies to mouse MHC antigens. III. Hybridoma antibodies reacting to antigens of the H-2b haplotype reveal genetic control of isotype expression. J Immunol. 1981 Jan;126(1):317–321. [PubMed] [Google Scholar]
  18. Potter H., Weir L., Leder P. Enhancer-dependent expression of human kappa immunoglobulin genes introduced into mouse pre-B lymphocytes by electroporation. Proc Natl Acad Sci U S A. 1984 Nov;81(22):7161–7165. doi: 10.1073/pnas.81.22.7161. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Reyes A. A., Schöld M., Wallace R. B. The complete amino acid sequence of the murine transplantation antigen H-2Db as deduced by molecular cloning. Immunogenetics. 1982;16(1):1–9. doi: 10.1007/BF00364437. [DOI] [PubMed] [Google Scholar]
  20. Ronne H., Widmark E., Rask L., Peterson P. A. Intron sequences reveal evolutionary relationships among major histocompatibility complex class I genes. Proc Natl Acad Sci U S A. 1985 Sep;82(17):5860–5864. doi: 10.1073/pnas.82.17.5860. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Rubin C. M., Schmid C. W. Pyrimidine-specific chemical reactions useful for DNA sequencing. Nucleic Acids Res. 1980 Oct 24;8(20):4613–4619. doi: 10.1093/nar/8.20.4613. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Sharrow S. O., Flaherty L., Sachs D. H. Serologic cross-reactivity between Class I MHC molecules and an H-2-linked differentiation antigen as detected by monoclonal antibodies. J Exp Med. 1984 Jan 1;159(1):21–40. doi: 10.1084/jem.159.1.21. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Sherman D. H., Kranz D. M., Eisen H. N. Expression of structurally diverse Qa-2-encoded molecules on the surface of cloned cytotoxic T lymphocytes. J Exp Med. 1984 Nov 1;160(5):1421–1430. doi: 10.1084/jem.160.5.1421. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Sim G. K., Yagüe J., Nelson J., Marrack P., Palmer E., Augustin A., Kappler J. Primary structure of human T-cell receptor alpha-chain. Nature. 1984 Dec 20;312(5996):771–775. doi: 10.1038/312771a0. [DOI] [PubMed] [Google Scholar]
  25. Soloski M. J., Vernachio J., Einhorn G., Lattimore A. Qa gene expression: biosynthesis and secretion of Qa-2 molecules in activated T cells. Proc Natl Acad Sci U S A. 1986 May;83(9):2949–2953. doi: 10.1073/pnas.83.9.2949. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Steinmetz M., Moore K. W., Frelinger J. G., Sher B. T., Shen F. W., Boyse E. A., Hood L. A pseudogene homologous to mouse transplantation antigens: transplantation antigens are encoded by eight exons that correlate with protein domains. Cell. 1981 Sep;25(3):683–692. doi: 10.1016/0092-8674(81)90175-6. [DOI] [PubMed] [Google Scholar]
  27. Stephan D., Sun H., Lindahl K. F., Meyer E., Hämmerling G., Hood L., Steinmetz M. Organization and evolution of D region class I genes in the mouse major histocompatibility complex. J Exp Med. 1986 May 1;163(5):1227–1244. doi: 10.1084/jem.163.5.1227. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Straus D. S., Stroynowski I., Schiffer S. G., Hood L. Expression of hybrid class I genes of the major histocompatibility complex in mouse L cells. Proc Natl Acad Sci U S A. 1985 Sep;82(18):6245–6249. doi: 10.1073/pnas.82.18.6245. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Stroynowski I., Forman J., Goodenow R. S., Schiffer S. G., McMillan M., Sharrow S. O., Sachs D. H., Hood L. Expression and T cell recognition of hybrid antigens with amino-terminal domains encoded by Qa-2 region of major histocompatibility complex and carboxyl termini of transplantation antigens. J Exp Med. 1985 May 1;161(5):935–952. doi: 10.1084/jem.161.5.935. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Vasseur M., Condamine H., Duprey P. RNAs containing B2 repeated sequences are transcribed in the early stages of mouse embryogenesis. EMBO J. 1985 Jul;4(7):1749–1753. doi: 10.1002/j.1460-2075.1985.tb03846.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Weiss E. H., Golden L., Fahrner K., Mellor A. L., Devlin J. J., Bullman H., Tiddens H., Bud H., Flavell R. A. Organization and evolution of the class I gene family in the major histocompatibility complex of the C57BL/10 mouse. Nature. 1984 Aug 23;310(5979):650–655. doi: 10.1038/310650a0. [DOI] [PubMed] [Google Scholar]
  32. Weiss E., Golden L., Zakut R., Mellor A., Fahrner K., Kvist S., Flavell R. A. The DNA sequence of the H-2kb gene: evidence for gene conversion as a mechanism for the generation of polymorphism in histocompatibilty antigens. EMBO J. 1983;2(3):453–462. doi: 10.1002/j.1460-2075.1983.tb01444.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Wigler M., Pellicer A., Silverstein S., Axel R., Urlaub G., Chasin L. DNA-mediated transfer of the adenine phosphoribosyltransferase locus into mammalian cells. Proc Natl Acad Sci U S A. 1979 Mar;76(3):1373–1376. doi: 10.1073/pnas.76.3.1373. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Wood W. I., Gitschier J., Lasky L. A., Lawn R. M. Base composition-independent hybridization in tetramethylammonium chloride: a method for oligonucleotide screening of highly complex gene libraries. Proc Natl Acad Sci U S A. 1985 Mar;82(6):1585–1588. doi: 10.1073/pnas.82.6.1585. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Zuniga M. C., Hood L. E. Clonal variation in cell surface display of an H-2 protein lacking a cytoplasmic tail. J Cell Biol. 1986 Jan;102(1):1–10. doi: 10.1083/jcb.102.1.1. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. van den Elsen P., Shepley B. A., Borst J., Coligan J. E., Markham A. F., Orkin S., Terhorst C. Isolation of cDNA clones encoding the 20K T3 glycoprotein of human T-cell receptor complex. 1984 Nov 29-Dec 5Nature. 312(5993):413–418. doi: 10.1038/312413a0. [DOI] [PubMed] [Google Scholar]

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