Skip to main content
NCBI home page
Nucleic Acids Research logoLink to Nucleic Acids Research
. 1992 Sep 11;20(17):4525–4531. doi: 10.1093/nar/20.17.4525

Ligand dependence of estrogen receptor induced changes in chromatin structure.

D M Gilbert 1, R Losson 1, P Chambon 1
PMCID: PMC334180  PMID: 1408752

Abstract

To determine whether the human estrogen receptor requires ligand to bind to its cognate estrogen receptor element (ERE) in vivo, we have examined the structure of chromatin at a chromosomally integrated ERE-URA3 reporter gene in yeast, and the influence of ligand bound and ligand free estrogen receptors on that structure. Using indirect end-labelling to map DNaseI and micrococcal nuclease sensitive sites, we found that receptor induced alterations in chromatin structure were completely dependent upon the presence of estradiol. These same alterations in chromatin structure were induced by a truncated estrogen receptor with both TAF-1 and TAF-2 transactivation functions deleted, suggesting that DNA binding per se disrupts chromatin structure. These results support models in which the estrogen receptor requires ligand to bind to the ERE in vivo.

Full text

PDF
4525

Images in this article

4528Image
on p.4528
4528Image
on p.4528
4529Image
on p.4529

Selected References

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

  1. Becker P. B., Gloss B., Schmid W., Strähle U., Schütz G. In vivo protein-DNA interactions in a glucocorticoid response element require the presence of the hormone. Nature. 1986 Dec 18;324(6098):686–688. doi: 10.1038/324686a0. [DOI] [PubMed] [Google Scholar]
  2. Bellard M., Dretzen G., Giangrande A., Ramain P. Nuclease digestion of transcriptionally active chromatin. Methods Enzymol. 1989;170:317–346. doi: 10.1016/0076-6879(89)70054-9. [DOI] [PubMed] [Google Scholar]
  3. Bradford M. M. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem. 1976 May 7;72:248–254. doi: 10.1016/0003-2697(76)90527-3. [DOI] [PubMed] [Google Scholar]
  4. Burch J. B., Weintraub H. Temporal order of chromatin structural changes associated with activation of the major chicken vitellogenin gene. Cell. 1983 May;33(1):65–76. doi: 10.1016/0092-8674(83)90335-5. [DOI] [PubMed] [Google Scholar]
  5. Cordingley M. G., Riegel A. T., Hager G. L. Steroid-dependent interaction of transcription factors with the inducible promoter of mouse mammary tumor virus in vivo. Cell. 1987 Jan 30;48(2):261–270. doi: 10.1016/0092-8674(87)90429-6. [DOI] [PubMed] [Google Scholar]
  6. Fedor M. J., Lue N. F., Kornberg R. D. Statistical positioning of nucleosomes by specific protein-binding to an upstream activating sequence in yeast. J Mol Biol. 1988 Nov 5;204(1):109–127. doi: 10.1016/0022-2836(88)90603-1. [DOI] [PubMed] [Google Scholar]
  7. Futcher A. B., Cox B. S. Copy number and the stability of 2-micron circle-based artificial plasmids of Saccharomyces cerevisiae. J Bacteriol. 1984 Jan;157(1):283–290. doi: 10.1128/jb.157.1.283-290.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Green S., Chambon P. Nuclear receptors enhance our understanding of transcription regulation. Trends Genet. 1988 Nov;4(11):309–314. doi: 10.1016/0168-9525(88)90108-4. [DOI] [PubMed] [Google Scholar]
  9. Gross D. S., Garrard W. T. Nuclease hypersensitive sites in chromatin. Annu Rev Biochem. 1988;57:159–197. doi: 10.1146/annurev.bi.57.070188.001111. [DOI] [PubMed] [Google Scholar]
  10. Han M., Grunstein M. Nucleosome loss activates yeast downstream promoters in vivo. Cell. 1988 Dec 23;55(6):1137–1145. doi: 10.1016/0092-8674(88)90258-9. [DOI] [PubMed] [Google Scholar]
  11. Kumar V., Chambon P. The estrogen receptor binds tightly to its responsive element as a ligand-induced homodimer. Cell. 1988 Oct 7;55(1):145–156. doi: 10.1016/0092-8674(88)90017-7. [DOI] [PubMed] [Google Scholar]
  12. Kumar V., Green S., Stack G., Berry M., Jin J. R., Chambon P. Functional domains of the human estrogen receptor. Cell. 1987 Dec 24;51(6):941–951. doi: 10.1016/0092-8674(87)90581-2. [DOI] [PubMed] [Google Scholar]
  13. Metzger D., White J. H., Chambon P. The human oestrogen receptor functions in yeast. Nature. 1988 Jul 7;334(6177):31–36. doi: 10.1038/334031a0. [DOI] [PubMed] [Google Scholar]
  14. Murdoch F. E., Gorski J. The role of ligand in estrogen receptor regulation of gene expression. Mol Cell Endocrinol. 1991 Jul;78(3):C103–C108. doi: 10.1016/0303-7207(91)90114-8. [DOI] [PubMed] [Google Scholar]
  15. Murdoch F. E., Meier D. A., Furlow J. D., Grunwald K. A., Gorski J. Estrogen receptor binding to a DNA response element in vitro is not dependent upon estradiol. Biochemistry. 1990 Sep 11;29(36):8377–8385. doi: 10.1021/bi00488a026. [DOI] [PubMed] [Google Scholar]
  16. Pham T. A., Elliston J. F., Nawaz Z., McDonnell D. P., Tsai M. J., O'Malley B. W. Antiestrogen can establish nonproductive receptor complexes and alter chromatin structure at target enhancers. Proc Natl Acad Sci U S A. 1991 Apr 15;88(8):3125–3129. doi: 10.1073/pnas.88.8.3125. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Pham T. A., Hwung Y. P., McDonnell D. P., O'Malley B. W. Transactivation functions facilitate the disruption of chromatin structure by estrogen receptor derivatives in vivo. J Biol Chem. 1991 Sep 25;266(27):18179–18187. [PubMed] [Google Scholar]
  18. Pratt W. B. Interaction of hsp90 with steroid receptors: organizing some diverse observations and presenting the newest concepts. Mol Cell Endocrinol. 1990 Nov 12;74(1):C69–C76. doi: 10.1016/0303-7207(90)90198-h. [DOI] [PubMed] [Google Scholar]
  19. Rigaud G., Roux J., Pictet R., Grange T. In vivo footprinting of rat TAT gene: dynamic interplay between the glucocorticoid receptor and a liver-specific factor. Cell. 1991 Nov 29;67(5):977–986. doi: 10.1016/0092-8674(91)90370-e. [DOI] [PubMed] [Google Scholar]
  20. Roy A., Exinger F., Losson R. cis- and trans-acting regulatory elements of the yeast URA3 promoter. Mol Cell Biol. 1990 Oct;10(10):5257–5270. doi: 10.1128/mcb.10.10.5257. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Thoma F. Protein-DNA interactions and nuclease-sensitive regions determine nucleosome positions on yeast plasmid chromatin. J Mol Biol. 1986 Jul 20;190(2):177–190. doi: 10.1016/0022-2836(86)90291-3. [DOI] [PubMed] [Google Scholar]
  22. Tora L., White J., Brou C., Tasset D., Webster N., Scheer E., Chambon P. The human estrogen receptor has two independent nonacidic transcriptional activation functions. Cell. 1989 Nov 3;59(3):477–487. doi: 10.1016/0092-8674(89)90031-7. [DOI] [PubMed] [Google Scholar]
  23. Wu C. Analysis of hypersensitive sites in chromatin. Methods Enzymol. 1989;170:269–289. doi: 10.1016/0076-6879(89)70052-5. [DOI] [PubMed] [Google Scholar]

Articles from Nucleic Acids Research are provided here courtesy of Oxford University Press

RESOURCES