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. 1990 Oct;10(10):5150–5159. doi: 10.1128/mcb.10.10.5150

Characterization of promoter elements of an interferon-inducible Ly-6E/A differentiation antigen, which is expressed on activated T cells and hematopoietic stem cells.

K D Khan 1, G Lindwall 1, S E Maher 1, A L Bothwell 1
PMCID: PMC361189  PMID: 1697928

Abstract

The Ly-6E/A antigen is expressed on activated murine T cells. Using probes made from the previously characterized cDNA, we have isolated a genomic DNA clone encoding the Ly-6A antigen. We determined the DNA sequence of the genomic clone and conducted a functional analysis of the promoter region. Mouse fibroblast BALB/3T3 cells transfected with this genomic clone constitutively expressed Ly-6A antigen on their cell surface. This expression was inducible by alpha/beta and gamma interferons. The Ly-6E 5'-flanking region was analyzed by chloramphenicol acetyltransferase assays in fibroblast cells for cis-acting elements. At least two positive elements were found to be needed for maximum constitutive promoter activity in L cells. One of the positive elements was specifically bound by a CCAAT box-binding protein from crude nuclear extract, as shown by electrophoretic mobility shift assays and footprinting. The other element, which contains a GGAAA motif and has homology to various known enhancers, also showed a specific binding activity. This second positive element when multimerized became a very powerful enhancing element. Interferon treatment could enhance expression of the chloramphenicol acetyltransferase gene fused to the Ly-6E 5'-flanking region in stably transfected BALB/3T3 cells. The elements responsible for this enhancement lie, at least in part, between positions -1760 and -900 of the gene. Surprisingly, there is no sequence homology between this region of Ly-6E and the established consensus for the interferon-stimulated response element, which has been shown functionally important to all previously characterized alpha/beta interferon-inducible promoters. The Ly-6E gene may prove to be a novel system for the study of interferon induction.

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Selected References

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  1. Alwine J. C. Transient gene expression control: effects of transfected DNA stability and trans-activation by viral early proteins. Mol Cell Biol. 1985 May;5(5):1034–1042. doi: 10.1128/mcb.5.5.1034. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Baldwin A. S., Jr, Sharp P. A. Binding of a nuclear factor to a regulatory sequence in the promoter of the mouse H-2Kb class I major histocompatibility gene. Mol Cell Biol. 1987 Jan;7(1):305–313. doi: 10.1128/mcb.7.1.305. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Ballard D. W., Philbrick W. M., Bothwell A. L. Identification of a novel 9-kDa polypeptide from nuclear extracts. DNA binding properties, primary structure, and in vitro expression. J Biol Chem. 1988 Jun 15;263(17):8450–8457. [PubMed] [Google Scholar]
  4. Borrelli E., Heyman R., Hsi M., Evans R. M. Targeting of an inducible toxic phenotype in animal cells. Proc Natl Acad Sci U S A. 1988 Oct;85(20):7572–7576. doi: 10.1073/pnas.85.20.7572. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Bothwell A., Pace P. E., LeClair K. P. Isolation and expression of an IFN-responsive Ly-6C chromosomal gene. J Immunol. 1988 Apr 15;140(8):2815–2820. [PubMed] [Google Scholar]
  6. Chodosh L. A., Baldwin A. S., Carthew R. W., Sharp P. A. Human CCAAT-binding proteins have heterologous subunits. Cell. 1988 Apr 8;53(1):11–24. doi: 10.1016/0092-8674(88)90483-7. [DOI] [PubMed] [Google Scholar]
  7. Cohen B., Vaiman D., Chebath J. Enhancer functions and in vitro protein binding of native and mutated interferon-responsive sequences. Nucleic Acids Res. 1989 Feb 25;17(4):1679–1695. doi: 10.1093/nar/17.4.1679. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Dignam J. D., Lebovitz R. M., Roeder R. G. Accurate transcription initiation by RNA polymerase II in a soluble extract from isolated mammalian nuclei. Nucleic Acids Res. 1983 Mar 11;11(5):1475–1489. doi: 10.1093/nar/11.5.1475. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Dorn A., Bollekens J., Staub A., Benoist C., Mathis D. A multiplicity of CCAAT box-binding proteins. Cell. 1987 Sep 11;50(6):863–872. doi: 10.1016/0092-8674(87)90513-7. [DOI] [PubMed] [Google Scholar]
  10. Felgner P. L., Gadek T. R., Holm M., Roman R., Chan H. W., Wenz M., Northrop J. P., Ringold G. M., Danielsen M. Lipofection: a highly efficient, lipid-mediated DNA-transfection procedure. Proc Natl Acad Sci U S A. 1987 Nov;84(21):7413–7417. doi: 10.1073/pnas.84.21.7413. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Fried M., Crothers D. M. Equilibria and kinetics of lac repressor-operator interactions by polyacrylamide gel electrophoresis. Nucleic Acids Res. 1981 Dec 11;9(23):6505–6525. doi: 10.1093/nar/9.23.6505. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Friedman R. L., Stark G. R. alpha-Interferon-induced transcription of HLA and metallothionein genes containing homologous upstream sequences. Nature. 1985 Apr 18;314(6012):637–639. doi: 10.1038/314637a0. [DOI] [PubMed] [Google Scholar]
  13. Fujita T., Shibuya H., Ohashi T., Yamanishi K., Taniguchi T. Regulation of human interleukin-2 gene: functional DNA sequences in the 5' flanking region for the gene expression in activated T lymphocytes. Cell. 1986 Aug 1;46(3):401–405. doi: 10.1016/0092-8674(86)90660-4. [DOI] [PubMed] [Google Scholar]
  14. Gorman C. M., Moffat L. F., Howard B. H. Recombinant genomes which express chloramphenicol acetyltransferase in mammalian cells. Mol Cell Biol. 1982 Sep;2(9):1044–1051. doi: 10.1128/mcb.2.9.1044. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. LeClair K. P., Bridgett M. M., Dumont F. J., Palfree R. G., Hämmerling U., Bothwell A. L. Kinetic analysis of Ly-6 gene induction in a T lymphoma by interferons and interleukin 1, and demonstration of Ly-6 inducibility in diverse cell types. Eur J Immunol. 1989 Jul;19(7):1233–1239. doi: 10.1002/eji.1830190713. [DOI] [PubMed] [Google Scholar]
  16. LeClair K. P., Palfree R. G., Flood P. M., Hammerling U., Bothwell A. Isolation of a murine Ly-6 cDNA reveals a new multigene family. EMBO J. 1986 Dec 1;5(12):3227–3234. doi: 10.1002/j.1460-2075.1986.tb04633.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Maity S. N., Golumbek P. T., Karsenty G., de Crombrugghe B. Selective activation of transcription by a novel CCAAT binding factor. Science. 1988 Jul 29;241(4865):582–585. doi: 10.1126/science.3399893. [DOI] [PubMed] [Google Scholar]
  18. Malek T. R., Ortega G., Chan C., Kroczek R. A., Shevach E. M. Role of Ly-6 in lymphocyte activation. II. Induction of T cell activation by monoclonal anti-Ly-6 antibodies. J Exp Med. 1986 Sep 1;164(3):709–722. doi: 10.1084/jem.164.3.709. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Palfree R. G., LeClair K. P., Bothwell A., Hämmerling U. cDNA characterization of an Ly-6.2 gene expressed in BW5147 tumor cells. Immunogenetics. 1987;26(6):389–391. doi: 10.1007/BF00343712. [DOI] [PubMed] [Google Scholar]
  20. Porter A. C., Chernajovsky Y., Dale T. C., Gilbert C. S., Stark G. R., Kerr I. M. Interferon response element of the human gene 6-16. EMBO J. 1988 Jan;7(1):85–92. doi: 10.1002/j.1460-2075.1988.tb02786.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Reiser H., Coligan J., Palmer E., Benacerraf B., Rock K. L. Cloning and expression of a cDNA for the T-cell-activating protein TAP. Proc Natl Acad Sci U S A. 1988 Apr;85(7):2255–2259. doi: 10.1073/pnas.85.7.2255. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Rock K. L., Reiser H., Bamezai A., McGrew J., Benacerraf B. The LY-6 locus: a multigene family encoding phosphatidylinositol-anchored membrane proteins concerned with T-cell activation. Immunol Rev. 1989 Oct;111:195–224. doi: 10.1111/j.1600-065x.1989.tb00547.x. [DOI] [PubMed] [Google Scholar]
  23. Spangrude G. J., Heimfeld S., Weissman I. L. Purification and characterization of mouse hematopoietic stem cells. Science. 1988 Jul 1;241(4861):58–62. doi: 10.1126/science.2898810. [DOI] [PubMed] [Google Scholar]
  24. Su B., Bothwell A. L. Biosynthesis of a phosphatidylinositol-glycan-linked membrane protein: signals for posttranslational processing of the Ly-6E antigen. Mol Cell Biol. 1989 Aug;9(8):3369–3376. doi: 10.1128/mcb.9.8.3369. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Tite J. P., Janeway C. A., Jr Antigen-dependent selection of B lymphoma cells varying in Ia density by cloned antigen-specific L3T4a+ T cells: a possible in vitro model for B cell adaptive differentiation. J Mol Cell Immunol. 1984;1(4):253–265. [PubMed] [Google Scholar]
  26. Tsang S. Y., Nakanishi M., Peterlin B. M. B-cell-specific and interferon-gamma-inducible regulation of the HLA-DR alpha gene. Proc Natl Acad Sci U S A. 1988 Nov;85(22):8598–8602. doi: 10.1073/pnas.85.22.8598. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Yeh E. T., Reiser H., Bamezai A., Rock K. L. TAP transcription and phosphatidylinositol linkage mutants are defective in activation through the T cell receptor. Cell. 1988 Mar 11;52(5):665–674. doi: 10.1016/0092-8674(88)90404-7. [DOI] [PubMed] [Google Scholar]
  28. van de Rijn M., Heimfeld S., Spangrude G. J., Weissman I. L. Mouse hematopoietic stem-cell antigen Sca-1 is a member of the Ly-6 antigen family. Proc Natl Acad Sci U S A. 1989 Jun;86(12):4634–4638. doi: 10.1073/pnas.86.12.4634. [DOI] [PMC free article] [PubMed] [Google Scholar]

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