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. 1989 Apr 11;17(7):2581–2595. doi: 10.1093/nar/17.7.2581

The contribution of the N- and C-terminal regions of steroid receptors to activation of transcription is both receptor and cell-specific.

M T Bocquel 1, V Kumar 1, C Stricker 1, P Chambon 1, H Gronemeyer 1
PMCID: PMC317644  PMID: 2717402

Abstract

Normalized dose response-curves for transcriptional activation of reporter genes were obtained by co-transfecting them with increasing amounts of wild-type (wt) progesterone (PR), glucocorticoid (GR) and oestrogen (ER) expression vectors. Marked differences in both shape and magnitude of the stimulation were observed depending on whether HeLa or CV1 cells were transfected. In HeLa cells the transcriptional stimulation from a reporter gene containing the hormone responsive element (RE) present in the mouse mammary tumour virus (MMTV) long terminal repeat (LTR) increased as increasing amounts (from 0.05 to 7.5 micrograms) of PR expression vector were transfected, whereas no such increase was observed in CV1 cells above 1 microgram of the same vector. In contrast, a PR mutant lacking the hormone binding domain (HBD, region E), exhibited increasing constitutive activity with increasing amounts of PR expression vector, such that in CV1 cells, but not in HeLa cells, similar activities were measured for the mutant and wt PR when 5 micrograms expression vectors were transfected. Western blot analyses indicated that the differences between the two cell lines were not due to differences in the amount of receptor proteins. Using the same MMTV LTR-based reporter gene, cell-specific differences were also detected between the dose-response curves obtained for the human GR and a mutant which lacks the HBD. A PR mutant in which the N-terminal A/B region was deleted exhibited no (CV1 cells) or less than 5% (HeLa cells) of the wt-activity, whereas the corresponding GR mutant stimulated efficiently transcription in both cell lines. Identical studies with the wt human ER or a mutant truncated for the N-terminal A/B region resulted in bell-shaped dose-response curves in both HeLa and CV1 cells, whereas an ER mutant lacking the HBD was weakly active in either cell line. These data demonstrate cell- and receptor-specificity for the transcriptional activation functions present in the A/B region and the HBD of various steroid receptors and suggest that limiting factors mediate their action. The present study also emphasizes the need of establishing dose-response curves to correctly assess the relative contribution of the different regions of steroid hormone receptors in activation of transcription.

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Selected References

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  1. Carson M. A., Tsai M. J., Conneely O. M., Maxwell B. L., Clark J. H., Dobson A. D., Elbrecht A., Toft D. O., Schrader W. T., O'Malley B. W. Structure-function properties of the chicken progesterone receptor A synthesized from complementary deoxyribonucleic acid. Mol Endocrinol. 1987 Nov;1(11):791–801. doi: 10.1210/mend-1-11-791. [DOI] [PubMed] [Google Scholar]
  2. Cato A. C., Miksicek R., Schütz G., Arnemann J., Beato M. The hormone regulatory element of mouse mammary tumour virus mediates progesterone induction. EMBO J. 1986 Sep;5(9):2237–2240. doi: 10.1002/j.1460-2075.1986.tb04490.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Danielsen M., Northrop J. P., Jonklaas J., Ringold G. M. Domains of the glucocorticoid receptor involved in specific and nonspecific deoxyribonucleic acid binding, hormone activation, and transcriptional enhancement. Mol Endocrinol. 1987 Nov;1(11):816–822. doi: 10.1210/mend-1-11-816. [DOI] [PubMed] [Google Scholar]
  4. Eul J., Meyer M. E., Tora L., Bocquel M. T., Quirin-Stricker C., Chambon P., Gronemeyer H. Expression of active hormone and DNA-binding domains of the chicken progesterone receptor in E. coli. EMBO J. 1989 Jan;8(1):83–90. doi: 10.1002/j.1460-2075.1989.tb03351.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Evans R. M. The steroid and thyroid hormone receptor superfamily. Science. 1988 May 13;240(4854):889–895. doi: 10.1126/science.3283939. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Giguère V., Hollenberg S. M., Rosenfeld M. G., Evans R. M. Functional domains of the human glucocorticoid receptor. Cell. 1986 Aug 29;46(5):645–652. doi: 10.1016/0092-8674(86)90339-9. [DOI] [PubMed] [Google Scholar]
  7. Gluzman Y. SV40-transformed simian cells support the replication of early SV40 mutants. Cell. 1981 Jan;23(1):175–182. doi: 10.1016/0092-8674(81)90282-8. [DOI] [PubMed] [Google Scholar]
  8. Godowski P. J., Picard D., Yamamoto K. R. Signal transduction and transcriptional regulation by glucocorticoid receptor-LexA fusion proteins. Science. 1988 Aug 12;241(4867):812–816. doi: 10.1126/science.3043662. [DOI] [PubMed] [Google Scholar]
  9. Godowski P. J., Rusconi S., Miesfeld R., Yamamoto K. R. Glucocorticoid receptor mutants that are constitutive activators of transcriptional enhancement. Nature. 1987 Jan 22;325(6102):365–368. doi: 10.1038/325365a0. [DOI] [PubMed] [Google Scholar]
  10. 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]
  11. Green S., Chambon P. A superfamily of potentially oncogenic hormone receptors. Nature. 1986 Dec 18;324(6098):615–617. doi: 10.1038/324615a0. [DOI] [PubMed] [Google Scholar]
  12. 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]
  13. Green S., Kumar V., Theulaz I., Wahli W., Chambon P. The N-terminal DNA-binding 'zinc finger' of the oestrogen and glucocorticoid receptors determines target gene specificity. EMBO J. 1988 Oct;7(10):3037–3044. doi: 10.1002/j.1460-2075.1988.tb03168.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Greene G. L., Sobel N. B., King W. J., Jensen E. V. Immunochemical studies of estrogen receptors. J Steroid Biochem. 1984 Jan;20(1):51–56. doi: 10.1016/0022-4731(84)90188-2. [DOI] [PubMed] [Google Scholar]
  15. Gronemeyer H., Govindan M. V., Chambon P. Immunological similarity between the chick oviduct progesterone receptor forms A and B. J Biol Chem. 1985 Jun 10;260(11):6916–6925. [PubMed] [Google Scholar]
  16. Gronemeyer H., Turcotte B., Quirin-Stricker C., Bocquel M. T., Meyer M. E., Krozowski Z., Jeltsch J. M., Lerouge T., Garnier J. M., Chambon P. The chicken progesterone receptor: sequence, expression and functional analysis. EMBO J. 1987 Dec 20;6(13):3985–3994. doi: 10.1002/j.1460-2075.1987.tb02741.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Guiochon-Mantel A., Loosfelt H., Ragot T., Bailly A., Atger M., Misrahi M., Perricaudet M., Milgrom E. Receptors bound to antiprogestin from abortive complexes with hormone responsive elements. Nature. 1988 Dec 15;336(6200):695–698. doi: 10.1038/336695a0. [DOI] [PubMed] [Google Scholar]
  18. Herbomel P., Bourachot B., Yaniv M. Two distinct enhancers with different cell specificities coexist in the regulatory region of polyoma. Cell. 1984 Dec;39(3 Pt 2):653–662. doi: 10.1016/0092-8674(84)90472-0. [DOI] [PubMed] [Google Scholar]
  19. Hollenberg S. M., Evans R. M. Multiple and cooperative trans-activation domains of the human glucocorticoid receptor. Cell. 1988 Dec 2;55(5):899–906. doi: 10.1016/0092-8674(88)90145-6. [DOI] [PubMed] [Google Scholar]
  20. Hollenberg S. M., Giguere V., Segui P., Evans R. M. Colocalization of DNA-binding and transcriptional activation functions in the human glucocorticoid receptor. Cell. 1987 Apr 10;49(1):39–46. doi: 10.1016/0092-8674(87)90753-7. [DOI] [PubMed] [Google Scholar]
  21. Hollenberg S. M., Giguere V., Segui P., Evans R. M. Colocalization of DNA-binding and transcriptional activation functions in the human glucocorticoid receptor. Cell. 1987 Apr 10;49(1):39–46. doi: 10.1016/0092-8674(87)90753-7. [DOI] [PubMed] [Google Scholar]
  22. Klein-Hitpass L., Schorpp M., Wagner U., Ryffel G. U. An estrogen-responsive element derived from the 5' flanking region of the Xenopus vitellogenin A2 gene functions in transfected human cells. Cell. 1986 Sep 26;46(7):1053–1061. doi: 10.1016/0092-8674(86)90705-1. [DOI] [PubMed] [Google Scholar]
  23. 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]
  24. Miesfeld R., Godowski P. J., Maler B. A., Yamamoto K. R. Glucocorticoid receptor mutants that define a small region sufficient for enhancer activation. Science. 1987 Apr 24;236(4800):423–427. doi: 10.1126/science.3563519. [DOI] [PubMed] [Google Scholar]
  25. Tora L., Gaub M. P., Mader S., Dierich A., Bellard M., Chambon P. Cell-specific activity of a GGTCA half-palindromic oestrogen-responsive element in the chicken ovalbumin gene promoter. EMBO J. 1988 Dec 1;7(12):3771–3778. doi: 10.1002/j.1460-2075.1988.tb03261.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Tora L., Gronemeyer H., Turcotte B., Gaub M. P., Chambon P. The N-terminal region of the chicken progesterone receptor specifies target gene activation. Nature. 1988 May 12;333(6169):185–188. doi: 10.1038/333185a0. [DOI] [PubMed] [Google Scholar]
  27. Webster N. J., Green S., Jin J. R., Chambon P. The hormone-binding domains of the estrogen and glucocorticoid receptors contain an inducible transcription activation function. Cell. 1988 Jul 15;54(2):199–207. doi: 10.1016/0092-8674(88)90552-1. [DOI] [PubMed] [Google Scholar]
  28. Yamamoto K. R. Steroid receptor regulated transcription of specific genes and gene networks. Annu Rev Genet. 1985;19:209–252. doi: 10.1146/annurev.ge.19.120185.001233. [DOI] [PubMed] [Google Scholar]

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