Skip to main content
The EMBO Journal logoLink to The EMBO Journal
. 1982;1(12):1613–1619. doi: 10.1002/j.1460-2075.1982.tb01363.x

Glucocorticoid receptors recognize DNA sequences in and around murine mammary tumour virus DNA.

S Geisse, C Scheidereit, H M Westphal, N E Hynes, B Groner, M Beato
PMCID: PMC553259  PMID: 6327283

Abstract

In several rodent cell lines, glucocorticoids increase the transcription of murine mammary tumour virus (MMTV) proviral DNA in a process mediated by the glucocorticoid receptor. To investigate whether a direct interaction between the receptor and specific sequences on the induced genes can be implicated in the hormonal regulation of transcription, filter binding studies were performed with partially purified glucocorticoid receptor of rat liver and eight cloned MMTV proviral probes. Both the 40 000 and the 90 00 mol. wt. forms of the receptor do bind preferentially to restriction fragments containing the right 400-500 nucleotides of the MMTV long terminal repeat units (LTR). Using LTR deletion mutants, we confirm that the right end of the LTR contains at least one binding site for the glucocorticoid receptor. In addition, the receptor binds preferentially to the mouse genomic sequences flanking at least three endogenous proviral copies, and to sequences within the env genes in some of the endogenous and exogenous proviruses. These findings are compatible with the hypothesis that steroid hormones regulate specific gene expression through a direct interaction of the hormone-receptor complex with DNA sequences in and around the induced genes.

Full text

PDF
1619

Images in this article

Selected References

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

  1. Buetti E., Diggelmann H. Cloned mouse mammary tumor virus DNA is biologically active in transfected mouse cells and its expression is stimulated by glucocorticoid hormones. Cell. 1981 Feb;23(2):335–345. doi: 10.1016/0092-8674(81)90129-x. [DOI] [PubMed] [Google Scholar]
  2. Carlstedt-Duke J., Okret S., Wrange O., Gustafsson J. A. Immunochemical analysis of the glucocorticoid receptor: identification of a third domain separate from the steroid-binding and DNA-binding domains. Proc Natl Acad Sci U S A. 1982 Jul;79(14):4260–4264. doi: 10.1073/pnas.79.14.4260. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Cochet M., Chang A. C., Cohen S. N. Characterization of the structural gene and putative 5'-regulatory sequences for human proopiomelanocortin. Nature. 1982 May 27;297(5864):335–339. doi: 10.1038/297335a0. [DOI] [PubMed] [Google Scholar]
  4. Dellweg H. G., Hotz A., Mugele K., Gehring U. Active domains in wild-type and mutant glucocorticoid receptors. EMBO J. 1982;1(3):285–289. doi: 10.1002/j.1460-2075.1982.tb01161.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Donehower L. A., Huang A. L., Hager G. L. Regulatory and coding potential of the mouse mammary tumor virus long terminal redundancy. J Virol. 1981 Jan;37(1):226–238. doi: 10.1128/jvi.37.1.226-238.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Fasel N., Pearson K., Buetti E., Diggelmann H. The region of mouse mammary tumor virus DNA containing the long terminal repeat includes a long coding sequence and signals for hormonally regulated transcription. EMBO J. 1982;1(1):3–7. doi: 10.1002/j.1460-2075.1982.tb01115.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Govindan M. V., 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 Sci U S A. 1982 Sep;79(17):5157–5161. doi: 10.1073/pnas.79.17.5157. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Gronemeyer H., Pongs O. Localization of ecdysterone on polytene chromosomes of Drosophila melanogaster. Proc Natl Acad Sci U S A. 1980 Apr;77(4):2108–2112. doi: 10.1073/pnas.77.4.2108. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Groner B., Buetti E., Diggelmann H., Hynes N. E. Characterization of endogenous and exogenous mouse mammary tumor virus proviral DNA with site-specific molecular clones. J Virol. 1980 Dec;36(3):734–745. doi: 10.1128/jvi.36.3.734-745.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Groner B., Hynes N. E. Number and location of mouse mammary tumor virus proviral DNA in mouse DNA of normal tissue and of mammary tumors. J Virol. 1980 Mar;33(3):1013–1025. doi: 10.1128/jvi.33.3.1013-1025.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Herrlich P., Hynes N. E., Ponta H., Rahmsdorf U., Kennedy N., Groner B. The endogenous proviral mouse mammary tumor virus genes of the GR mouse are not identical and only one corresponds to the exogenous virus. Nucleic Acids Res. 1981 Oct 10;9(19):4981–4995. doi: 10.1093/nar/9.19.4981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Huang A. L., Ostrowski M. C., Berard D., Hager G. L. Glucocorticoid regulation of the Ha-MuSV p21 gene conferred by sequences from mouse mammary tumor virus. Cell. 1981 Dec;27(2 Pt 1):245–255. doi: 10.1016/0092-8674(81)90408-6. [DOI] [PubMed] [Google Scholar]
  13. Hynes N. E., Kennedy N., Rahmsdorf U., Groner B. Hormone-responsive expression of an endogenous proviral gene of mouse mammary tumor virus after molecular cloning and gene transfer into cultured cells. Proc Natl Acad Sci U S A. 1981 Apr;78(4):2038–2042. doi: 10.1073/pnas.78.4.2038. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Kennedy N., Knedlitschek G., Groner B., Hynes N. E., Herrlich P., Michalides R., van Ooyen A. J. Long terminal repeats of endogenous mouse mammary tumour virus contain a long open reading frame which extends into adjacent sequences. Nature. 1982 Feb 18;295(5850):622–624. doi: 10.1038/295622a0. [DOI] [PubMed] [Google Scholar]
  15. Lee F., Mulligan R., Berg P., Ringold G. Glucocorticoids regulate expression of dihydrofolate reductase cDNA in mouse mammary tumour virus chimaeric plasmids. Nature. 1981 Nov 19;294(5838):228–232. doi: 10.1038/294228a0. [DOI] [PubMed] [Google Scholar]
  16. Michalides R., van Nie R., Nusse R., Hynes N. E., Groner B. Mammary tumor induction loci in GR and DBAf mice contain one provirus of the mouse mammary tumor virus. Cell. 1981 Jan;23(1):165–173. doi: 10.1016/0092-8674(81)90281-6. [DOI] [PubMed] [Google Scholar]
  17. Mulvihill E. R., LePennec J. P., Chambon P. Chicken oviduct progesterone receptor: location of specific regions of high-affinity binding in cloned DNA fragments of hormone-responsive genes. Cell. 1982 Mar;28(3):621–632. doi: 10.1016/0092-8674(82)90217-3. [DOI] [PubMed] [Google Scholar]
  18. Payvar F., Wrange O., Carlstedt-Duke J., Okret S., Gustafsson J. A., Yamamoto K. R. Purified glucocorticoid receptors bind selectively in vitro to a cloned DNA fragment whose transcription is regulated by glucocorticoids in vivo. Proc Natl Acad Sci U S A. 1981 Nov;78(11):6628–6632. doi: 10.1073/pnas.78.11.6628. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Riggs A. D., Suzuki H., Bourgeois S. Lac repressor-operator interaction. I. Equilibrium studies. J Mol Biol. 1970 Feb 28;48(1):67–83. doi: 10.1016/0022-2836(70)90219-6. [DOI] [PubMed] [Google Scholar]
  20. Ringold G. M. Glucocorticoid regulation of mouse mammary tumor virus gene expression. Biochim Biophys Acta. 1979 Dec 19;560(4):487–508. doi: 10.1016/0304-419x(79)90014-3. [DOI] [PubMed] [Google Scholar]
  21. Schaffner W., Weissmann C. A rapid, sensitive, and specific method for the determination of protein in dilute solution. Anal Biochem. 1973 Dec;56(2):502–514. doi: 10.1016/0003-2697(73)90217-0. [DOI] [PubMed] [Google Scholar]
  22. Westphal H. M., Beato M. Influence of pyridoxal 5'-phosphate on the DNA binding activity of steroid hormone receptors and other DNA binding proteins. FEBS Lett. 1981 Feb 23;124(2):189–192. doi: 10.1016/0014-5793(81)80133-0. [DOI] [PubMed] [Google Scholar]
  23. Westphal H. M., Beato M. The activated glucocorticoid receptor of rat liver. Purification and physical characterization. Eur J Biochem. 1980 May;106(2):395–403. doi: 10.1111/j.1432-1033.1980.tb04585.x. [DOI] [PubMed] [Google Scholar]
  24. Westphal H. M., Fleischmann G., Beato M. Photoaffinity labeling of steroid binding proteins with unmodified ligands. Eur J Biochem. 1981 Sep;119(1):101–106. doi: 10.1111/j.1432-1033.1981.tb05582.x. [DOI] [PubMed] [Google Scholar]
  25. Winter R. B., von Hippel P. H. Diffusion-driven mechanisms of protein translocation on nucleic acids. 2. The Escherichia coli repressor--operator interaction: equilibrium measurements. Biochemistry. 1981 Nov 24;20(24):6948–6960. doi: 10.1021/bi00527a029. [DOI] [PubMed] [Google Scholar]
  26. Wrange O., Carlstedt-Duke J., Gustafsson J. A. Purification of the glucocorticoid receptor from rat liver cytosol. J Biol Chem. 1979 Sep 25;254(18):9284–9290. [PubMed] [Google Scholar]

Articles from The EMBO Journal are provided here courtesy of Nature Publishing Group

RESOURCES