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Isolation and Sequence of the Human Glucocorticoid Receptor Gene Promoter. Cloning of the Human Androgen Receptor cDNA

  • Manjapra V. Govindan
  • Marco Burelle
  • Celine Cantin
  • Martine Devic
  • Claude Labrie
  • Fernand Labrie
  • Yves Lachance
  • Gilles Leblanc
  • Claude Lefebvre
  • Pravin Patel
  • Jacques Simard
  • Udo Stropp
Part of the Serono Symposia, USA book series (SERONOSYMP)

Abstract

We have isolated the promoter region of the human glucocorticoid receptor (hGR) gene from the λEMBL-3 human genomic library using synthetic oligonucleotides corresponding to the 5’ end of hGR cDNA. The clone was fine mapped by restriction digestion, hybridization with hGR cDNA probes and by DNA sequence analysis and found to contain an open reading frame corresponding to the hGR cDNA from amino acids 1–131 separated by a large intron from the 5’ noncoding sequences. The promoter region of the hGR has been identified by primer extension, nuclease S1 mapping using hGR mRNA from human prostatic carcinoma cells (LNCaP) and human breast tumor cells (MCF-7), by in vitro transcription using hGR-DNA fragments as template and by DNA sequence analysis. The sequence of the hGR promoter reveals that it contains neither a “TATA box” nor a “CART box” and has an extremely high “G+C” content. Gene transfer studies with hGR-CAT chimeric plasmids into COS-cells showed that the hormonal regulatory sequences of hGR is contained within a 4 kb EcoRl-XbaI fragment.

Keywords

Androgen Receptor Glucocorticoid Receptor LNCaP Cell Mouse Mammary Tumor Virus Androgen Receptor mRNA 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.

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References

  1. 1.
    Yamamoto KR. Steroid receptor regulated transcription of specific genes and gene networks. Annu Rev Genet 1985; 19: 209–52.PubMedCrossRefGoogle Scholar
  2. 2.
    Payvar F, Wrange O, Carlstedt-Duke J, et al. Purified glucocorticoid receptors bind selectively in vitro to a cloned DNA fragment whose transcription is regulated by glucocorticoids in vivo. Proc Natl Acad Sci USA 1981; 78: 6628–32.PubMedCrossRefGoogle Scholar
  3. 3.
    Govindan MV, Spiess E, Majors J. Purified glucocorticoid receptor-hormone complex from rat liver cytosol binds specifically to cloned mouse mammary tumor virus long-terminal repeats in vitro. Proc Natl Acad USA 1982; 17: 5157–61.CrossRefGoogle Scholar
  4. 4.
    Geisse S, Scheidereit C, Westphal HM, et al. Glucocorticoid receptors recognize DNA sequences in and around murine mammary tumour virus DNA. EMBO J 1982; 1: 1613–9.PubMedGoogle Scholar
  5. 5.
    Pfahl M. Specific binding of the glucocorticoid-receptor complex to the mouse mammary tumor proviral promoter region. Cell 1982; 31: 475–82.PubMedCrossRefGoogle Scholar
  6. 6.
    Scheidereit C, Geisse S, Westphal HM, et al. The glucocorticoid receptor binds to defined nucleotide sequences near the promoter of mouse mammary tumour virus. Nature 1983; 749–52.Google Scholar
  7. 7.
    Payvar F, De Franco D, Firestone GL, et al. Sequence-specific binding of glucocorticoid receptor to MTV DNA at sites within and upstream of the transcribed region. Cell 198; 35: 381–92.Google Scholar
  8. 8.
    Scheidereit C, Beato M. Contacts between hormone receptor and DNA double helix within a glucocorticoid regulatory element of mouse mammary tumor virus. Proc Natl Acad Sci USA 1984; 81: 3029–33.PubMedCrossRefGoogle Scholar
  9. 9.
    Karin M, Haslinger A, Holtgreve H, et al. Characterization of DNA sequences through which cadmium and glucocorticoid hormones induce human metallothionein-IIA gene. Nature 1984; 308: 513–9.PubMedCrossRefGoogle Scholar
  10. 10.
    Renhawitz R, Schutz G, von der Ahe D, et al. Sequences in the promoter region of the chicken lysozyme gene required for steroid regulation and receptor binding. Cell 1984; 37: 503–10.CrossRefGoogle Scholar
  11. 11.
    Liao S. Cellular receptors and mechanisms of action of steroid hormones. Int Rev Cytol 1975; 41: 87–172.PubMedCrossRefGoogle Scholar
  12. 12.
    Mooradian AD, Morley JE, Korenman L. Biological actions of androgens. Endocr Rev 1987; 1–28.Google Scholar
  13. 13.
    Smith RG, Nag A, Syms AJ, Norris JS. II. Steroid receptor, gene structure and molecular biology: steroid regulation of receptor concentration and oncogene expression. J Steroid Biochem 1986; 24: 51–5.PubMedCrossRefGoogle Scholar
  14. 14.
    Tindall DJ, Chang CH, Lobl TJ, Cunningham GR. Androgen antagonists in androgen target tissues. Pharmacol Ther 1984; 24: 367–400.PubMedCrossRefGoogle Scholar
  15. 15.
    Rowley DR, Tindall DJ. Androgen receptor protein: purification and molecular properties. In: Litwack G, ed. Biochemical actions of hormones; vol X III. New York: Academic Press, 1984.Google Scholar
  16. 16.
    Labrie F, Dupont A, Belanger A, et al. Treatment of prostate cancer with gonadotropin-releasing hormone agonists. Endocr Rev 1986; 7: 67–74.PubMedCrossRefGoogle Scholar
  17. 17.
    Labrie F, Luthy I, Veilleux R, Simard J, Belanger A, Dupont A. New concepts on the androgen sensitivity of prostate cancer. In: Murphy GP, Khoury S, Kuss R, Chatelain C, Denis L, eds. New York: Alan R. Liss, Inc., 1987: 145–52.Google Scholar
  18. 18.
    Labrie F, Dupont A, Giguere M, et al. Advantages of the combination therapy in previously untreated and treated patients with advanced prostate cancer. J Steroid Biochem 1986; 25: 877–83.PubMedCrossRefGoogle Scholar
  19. 19.
    Bardin CW, Catterall JF. Testosterone: a major determinant of extragenital sexual dimorphism. Science 1981; 211: 1285–94.PubMedCrossRefGoogle Scholar
  20. 20.
    Keenan BS, Meyer WJ, Hadjian AJ, Jones HW, Migeon CJ. Syndrome of androgen insensitivity in man: absence of 5-alpha-dihydrotestosterone-binding protein in skin fibroblasts. J Clin Endocrinol Metab 1974; 38: 1143–6.PubMedCrossRefGoogle Scholar
  21. 21.
    Meyer WJ, Migeon BR, Migeon CJ. Locus of human X chromosome for dihydrotestosterone receptor and androgen insensitivity. Proc Natl Acad Sci USA 1975; 72: 1469–72.PubMedCrossRefGoogle Scholar
  22. 22.
    Wilson JD, George FW, Griffin JE. The hormonal control of sexual development. Science 1981; 211: 1278–84.PubMedCrossRefGoogle Scholar
  23. 23.
    Chang CM, Rowley DR, Lobl TJ, Tindall DJ. Purification and characterization of androgen receptor from steer seminal vesicle. Biochemistry 1982; 4102–9.Google Scholar
  24. 24.
    Chang CM, Tindall DJ. Physiochemical characterization of the androgen receptor in rat uterine cytosol. Endocrinology 1983; 113: 148–693.CrossRefGoogle Scholar
  25. 25.
    Rowley DR, Chang CM, Tindall DJ. Effects of sodium molybdate on the androgen receptor from the R3327 prostatic tumor. Endocrinology 1984; 114: 1776–83.PubMedCrossRefGoogle Scholar
  26. 26.
    Asselin J, Melancon R, Moachon G, Belanger A. Characteristics of binding to estrogen, androgen, progestin and glucocorticoid receptors in 7,12-dimethylbenz(a)anthracene-induced mammary tumors and their hormonal control. Cancer Res 1980; 40: 1612–22.PubMedGoogle Scholar
  27. 27.
    Barnert JM, Jeltsch JM, Staub A, Jensen E, Scrace G, Waterfield M, Chambon P. Cloning of the human estrogen receptor cDNA. Proc Natl Acad Sci USA 1985; 82: 7889–93.CrossRefGoogle Scholar
  28. 28.
    Hollenberg SM, Weinberger C, Ong ES, et al. Primary structure and expression of a functional human glucocorticoid receptor cDNA. Nature 1985; 318: 635–41.PubMedCrossRefGoogle Scholar
  29. 29.
    Govindan MV, Devic M, Green S, Gonemeyer H, Chambon P. Cloning of the human glucocorticoid receptor cDNA. Nucleic Acids Res 1985; 13: 8293–304.PubMedCrossRefGoogle Scholar
  30. 30.
    Misrahi M, Atger M, D’Auriol L, et al. Complete amino acid sequence of the human progesterone receptor deduced from cloned cDNA. Biochem Biophys Res Commun 1987; 143: 740–8.PubMedCrossRefGoogle Scholar
  31. 31.
    Arriza JL, Weinberg C, Cerelli G, et al. Cloning of human mineralocorticoid receptor complementary DNA: structural and functional kinship with the glucocorticoid receptor. Science 1987; 237: 268–75.PubMedCrossRefGoogle Scholar
  32. 32.
    Govindan MV, Burelle M, Cantin C, et al. Cloning of human androgen receptor cDNA. Proc of the Int Conf on Hormones and Cancer, Hamburg, 1987.Google Scholar
  33. 33.
    Miller J, McLachlan AD, Klug A. Repetitive zinc-binding domains in the protein transcription factor III-A from Xenopus oocytes. EMBO J 1985; 41: 1609–14.Google Scholar
  34. 34.
    Sanger F, Nicklen S, Coulson AR. DNA sequencing with chain terminating inhibitors. Proc Natl Acad Sci USA; 74: 5463–7.Google Scholar
  35. 35.
    Lemeure M, Glandville N, Mandel JL, Gerlinger P, Palmiter R, Chambon P. The ovalbumin gene family: hormonal control of X and Y gene transcription and mRNA accumulation. Cell 1981: 23: 561–71.CrossRefGoogle Scholar
  36. 36.
    Nelsen C, Crenshaw EB III, Franco R, Lira SA, Alber VR, Evans RM, Rosenfeld MV. Discrete cis-active genomic sequences dictate the pituitary cell type-specific expression of rate prolactin and growth hormone genes. Nature 1986; 322: 557–62.CrossRefGoogle Scholar
  37. 37.
    Davis GL, Dibner MD, Battey JF. Basic methods in molecular biology. New York: Elsevier, 1986.Google Scholar
  38. 38.
    Weingartener B, Keller W. Transcription and processing of adenoviral RNA by extracts from Hela cells. Proc Natl Acad Sci USA 1981; 78: 4092–6.CrossRefGoogle Scholar
  39. 39.
    Maniatis T, Fritsch EF, Sambrook J. Molecular cloning. Cold Spring Harbor Laboratory.Google Scholar
  40. 40.
    Laemmli UK. Cleavage of structural proteins during the assembly of the head of the bacteriophage T4. Nature; 227: 680.Google Scholar
  41. 41.
    Southern E. Detection of specific sequences among DNA fragments separated by gel electrophoresis. J Mol Biol 1975; 98: 503.PubMedCrossRefGoogle Scholar
  42. 42.
    Frischauf AM, et al. Lambda replacement vectors carrying polylinker sequences. J Mol Biol 1983; 170: 827–42.PubMedCrossRefGoogle Scholar
  43. 43.
    Breathnach R, Chambon P. Organization and expression of eucaryotic split genes coding for proteins. Annu Rev Biochem 1981; 50: 349–83.PubMedCrossRefGoogle Scholar
  44. 44.
    Fromm M, Berg P. Deletion mapping of DNA regions required for SV40 early region promoter function in vivo. J Mol Appl Genet 1982; 1: 457–81.PubMedGoogle Scholar
  45. 45.
    Ishii S, Xu Y-H, Stratton RH, et al. Characterization and sequence of the promoter region of the human epidermal growth factor receptor gene. Proc Natl Acad Sci USA 1985; 82: 4920–4.PubMedCrossRefGoogle Scholar
  46. 46.
    Benoist C, Chambon P. In vivo sequence requirement of the SV40 early promoter region. Nature 1981; 290: 304–10.PubMedCrossRefGoogle Scholar
  47. 47.
    Hansen U, Sharp PA. Sequences controlling in vitro transcription of SV40 promoters. EMBO J 1983; 2: 2293–303.PubMedGoogle Scholar
  48. 48.
    Wasylyk B, Chambon P. Potentiator effect of the SV40 72-bp repeat on initiation of transcription from heterologous promoter elements. Cold Spring Harbor Symposium. Quant Biol; 47: 921–34.Google Scholar
  49. 49.
    Tjian R. T antigen binding and the control of SV40 gene expression. Cell 1981; 26: 1–2.PubMedCrossRefGoogle Scholar
  50. 50.
    Reynolds GA, Basu SK, Osborne TH, et al. HMG CoA reductase: a negatively regulated gene with unusual promoter and 5’-untranslated regions. Cell 1984; 38: 275–85.PubMedCrossRefGoogle Scholar
  51. 51.
    Serfling E, Jasin M, Shaffner W. Enhancers and eukaryotic gene transcription. Trends in genetics. August 1985, 224–30.Google Scholar
  52. 52.
    Gidoni D, Dynan WS, Tjian R. Multiple specific contacts between a mammalian transcription factor and its cognate promoters. Nature 1984; 312: 409–13.PubMedCrossRefGoogle Scholar
  53. 53.
    Dynan WS, Tjian R. Isolation of transcription factors that discriminate between different promoters recognized by RNA polymerase II. Cell 1983; 32: 669–80.PubMedCrossRefGoogle Scholar
  54. 54.
    Okret S, Poellinger L, Dong Y, Gustafsson JA. Down-regulation of glucocorticoid receptor mRNA by glucocorticoid hormones and recognition by the receptor of a specific binding sequence within a receptor cDNA clone. Proc Natl Acad Sci USA 1986; 83: 5899–903.PubMedCrossRefGoogle Scholar
  55. 55.
    Gorman MC, Moffat FL, Howard BH. Recombinant genomes which express chloramphenicol acetyltransferase in mammalian cells. Mol Cell Biol 1982; (Sep):1044–51.Google Scholar
  56. 56.
    Miefsfeld R, Ruscini S, Godowski PJ, et al. Genetic complementation of a glucocorticoid receptor deficiency by expression of cloned receptor cDNA. Cell 1986; 46: 389–99.CrossRefGoogle Scholar
  57. 57.
    Asselin J, Labrie F, Gourdeau Y, Bonne C, Raynaud JP. Binding of [H]methyltrienolone (R1881) in rat prostate and human benign prostatic hypertrophy (BPH). Steroids 1976; 28: 449–59.PubMedCrossRefGoogle Scholar
  58. 58.
    Simard J, Luthy I, Guay J, Belanger A, Labrie F. Characteristics of interaction of the antiandrogen flutamide with androgen receptor in various target tissues. Mol Cell Endocrinol 1986; 44: 261–70.PubMedCrossRefGoogle Scholar

Copyright information

© Plenum Press, New York 1988

Authors and Affiliations

  • Manjapra V. Govindan
    • 1
  • Marco Burelle
    • 1
  • Celine Cantin
    • 1
  • Martine Devic
    • 2
  • Claude Labrie
    • 1
  • Fernand Labrie
    • 1
  • Yves Lachance
    • 1
  • Gilles Leblanc
    • 1
  • Claude Lefebvre
    • 1
  • Pravin Patel
    • 3
  • Jacques Simard
    • 1
  • Udo Stropp
    • 2
  1. 1.MRC Group in Molecular EndocrinologyLavai University Medical CenterCanada
  2. 2.L.G.M.E., C.N.R.S., U-184INSERM Institut de Biochimie BiologiqueStrasbourgFrance
  3. 3.Department of Microbiology, Faculty of MedicineLaval UniversityCanada

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