[en] Glucocorticoids have been shown to inhibit the activity of the human prolactin (hPRL) promoter. Using transient expression experiments in rat pituitary cells, we located the sequence conferring glucocorticoid inhibition to a region which contains Pit-1 binding sites, responsible for pituitary-specific expression, but does not seem to contain a glucocorticoid receptor (GR) binding site. Co-transfection experiments in non-pituitary cell lines, using expression vectors for Pit-1 and different mutants of the human GR show that inhibition of the hPRL gene is seen only in the presence of Pit-1 and GR, and that the DNA binding function of the receptor is not required. Immunoprecipitation studies show that either anti-GR or anti-Pit-1 antibodies are able to co-precipitate GR and Pit-1, suggesting an interaction between these factors. We conclude that the activated GR functionally interferes with the pituitary specific factor Pit-1, thereby leading to the observed transcriptional repression.
Disciplines :
Biochemistry, biophysics & molecular biology
Author, co-author :
Nalda, Asunción M; Université Paris XI
Martial, Joseph ; Université de Liège - ULiège > Département des sciences de la vie > GIGA-R : Biologie et génétique moléculaire
Muller, Marc ; Université de Liège - ULiège > Département des sciences de la vie > GIGA-R : Biologie et génétique moléculaire
Language :
English
Title :
The glucocorticoid receptor inhibits the human prolactin gene expression by interference with Pit-1 activity
Publication date :
1997
Journal title :
Molecular and Cellular Endocrinology
ISSN :
0303-7207
eISSN :
1872-8057
Publisher :
North Holland Publishing Company, Amsterdam, Netherlands
Adler, S., Waterman, M.L., He, X., Rosenfeld, M.G., Steroid receptor-mediated inhibition of rat prolactin gene expression does not require the receptor DNA-binding domain (1988) Cell, 52, pp. 685-695
Akerblom, J.E., Slater, E.P., Beato, M., Baxter, J.D., Mellon, P.L., Negative regulation by glucocorticoids through interference with a cAMP responsive element (1988) Science, 241, pp. 350-351
Berwaer, M., Monget, P., Peers, B., Mathy-Hartert, M., Bellefroid, E., Davis, J.R.E., Belayew, A., Martial, J.A., Multi-hormonal regulation of the human prolactin gene expression from 5000 bp of its upstream sequence (1991) Mol. Cell. Endocrinol., 80, pp. 53-64
Camper, S.A., Yao, Y.A.S., Rottman, F.M., Hormonal regulation of the bovine prolactin promoter in rat pituitary tumor cells (1985) J. Biol. Chem., 260, pp. 12246-12251
Crenshaw, E.B., Kall, K., Simmons, D.M., Swanson, L.W., Rosenfeld, M.G., Cell-specific expression of the prolactin gene in transgenic mice is controlled by synergistic interactions between promoter and enhancer elements (1989) Genes Dev., 3, pp. 959-972
Day, R.N., Maurer, R.A., The distal enhancer region of the rat prolactin gene contains elements conferring response to multiple hormones (1989) Mol. Endocrinol., 3, pp. 3-9
De Camili, P., MacConi, D., Spada, A., Dopamine inhibits adenylate cyclase in human prolactin-secreting pituitary adenomas (1979) Nature, 278, pp. 252-254
Drouin, J., Sun, Y.L., Chamberland, M., Gauthier, Y., De Lean, A., Nemer, M., Schmidt, T.J., Novel glucocorticoid receptor complex with DNA element of the hormone-repressed POMC gene (1993) EMBO J., 12, pp. 145-156
Eggert, M., Muller, M., Renkawitz, R., The Glucocorticoid Receptor (1995) Inducible Gene Expression, 2, pp. 131-156. , In: Bauerle, P.A. (Ed.), Birkhauser Verlag, Boston
Elsholtz, H.P., Lew, A.M., Albert, P.R., Sundmark, V.C., Inhibitory control of prolactin and Pit-1 gene promoters by dopamine (1991) J. Biol. Chem., 226, pp. 22919-22925
Fox, R.S., Jong, T.C., Casanova, J., Ye, Z.S., Stanley, F., Samuels, H.H., The homeodomain protein, Pit-1/GHF-1, is capable of binding to and activating cell-specific elements of both the growth hormone and prolactin gene promoters (1990) Mol. Endocrinol., 4, pp. 1069-1080
Giguère, V., Hollenberg, S.M., Rosenfeld, M.G., Evans, R.M., Functional domains of the human glucocorticoid receptor (1986) Cell, 46, pp. 645-652
Graham, F.L., Van Der Eb, A.J., A new technique for the assay of human adenovirus 5 DNA (1973) Virology, 52, pp. 456-457
Helmberg, A., Auphan, N., Caelles, C., Karin, M., Glucocorticoid-induced apoptosis of human leukemic cells is caused by the repressive function of the glucocorticoid receptor (1995) EMBO J., 3, pp. 452-460
Hoggard, N., Davis, J.R.E., Berwaer, M., Monget, P., Peers, B., Belayew, A., Martial, J.A., Pit-1 binding sequences permit calcium regulation of human prolactin gene expression (1991) Mol. Endocrinol., 5, pp. 1748-1754
Hollenberg, S.M., Evans, R.M., Multiple and cooperative trans-activation domains of the human glucocorticoid receptor (1988) Cell, 55, pp. 899-906
Horwitz, K.B., Costlow, M.E., McGuire, W.L., MCF-7: A human breast cancer cell line with estrogen, androgen, progesterone, and glucocorticoid receptors (1975) Steroids, 26, pp. 785-795
Jonat, C., Rahmsdorf, H.J., Park, K.-K., Cato, A.C.B., Gebel, S., Ponta, H., Herrlich, P., Anti-tumor promotion and anti-inflammation: Down-modulation of AP-1 (Fos/Jun) activity by glucocorticoid hormone (1990) Cell, 62, pp. 1189-1204
Lew, A.M., Elsholtz, H.P., Cloning of the human cDNA for transcription factor Pit-1 (1991) Nucleic Acids Res., 19, p. 6329
Mangalam, H.J., Albert, V.R., Ingraham, H.A., Kapiloff, M., Wilson, L., Nelson, C., Elsholtz, H., Rosenfeld, M.G., A pituitary POU domain protein, Pit-1, activates both growth hormone and prolactin promoters transcriptionally (1989) Genes Dev., 3, pp. 946-958
Murdoch, G.H., Waterman, M., Evans, R.M., Rosenfeld, M.G., Molecular mechanisms of phorbol esters, thyrotropin-releasing hormone, and growth factor stimulation of prolactin gene transcription (1985) J. Biol. Chem., 260, pp. 11852-11858
Peers, B., Voz, M.L., Monget, P., Mathy-Hartert, M., Berwaer, M., Belayew, A., Martial, J.A., Regulatory elements controlling pituitary-specific expression of the human prolactin gene (1990) Mol. Cell. Biol., 10, pp. 4690-4700
Peers, B., Monget, P., Nalda, A.M., Voz, M., Berwaer, M., Belayew, A., Martial, J.A., Transcriptional induction of the human prolactin gene by cAMP requires two cis-acting elements and at least the pituitary-specific factor Pit-1 (1991) J. Biol. Chem., 266, pp. 18127-18134
Ray, A., Prefontaine, K.E., Physical association and functional antagonism between the p65 subunit of transcription factor NF-kB and the glucocorticoid receptor (1994) Proc. Natl. Acad. Sci. USA, 91, pp. 752-756
Sakai, D.D., Helms, S., Carlstedt-Duke, J., Gustafsson, J.-A., Rottman, F.M., Yamamoto, K.R., Hormone-mediated repression: A negative glucocorticoid response element from the bovine prolactin gene (1988) Genes Dev., 2, pp. 1144-1154
Samuels, H.H., Stanley, F., Casanova, J., Depletion of L-3,5,3′-triiodothyronine and L-thyroxine in euthyroid calf serum for use in cell culture studies of the action of thyroid hormone (1979) Endocrinology, 105, pp. 80-85
Sanger, F., Nicklen, S., Coulson, A.R., DNA sequencing with chain-terminating inhibitors (1977) Proc. Natl. Acad. Sci. USA, 74, pp. 5463-5467
Schüle, R., Muller, M., Kaltschmidt, C., Renkawitz, R., Many transcription factors interact synergistically with steroid receptors (1988) Science, 242, pp. 1418-1420
Shapiro, D.J., Sharp, P.A., Wahli, W.W., Keller, M., A high-efficiency HeLa nuclear transcription extract (1988) DNA, 7, pp. 47-55
Subramaniam, N., Cairns, W., Okret, S., Studies on the mechanism of glucocorticoid-mediated repression from a negative glucocorticoid response element from the bovine prolactin gene (1997) DNA Cell Biol., 16, pp. 153-163
Yan, G.Z., Bancroft, C., Mediation by calcium of thyrotropin-releasing hormone action on the prolactin promoter via transcription factor Pit-1 (1991) Mol. Endocrinol., 5, pp. 1186-1189
Yan, G.Z., Pan, W.T., Bancroft, C., Thyrotropin-releasing hormone action on the prolactin promoter is mediated by the POU protein Pit-1 (1991) Mol. Endocrinol., 5, pp. 535-541
Yang-Yen, H.-F., Chambond, J.-C., Sun, Y.-L., Smeal, T., Schmidt, T.J., Drouin, J., Karin, M., Transcriptional interference between c-jun and the glucocorticoid receptor: Mutual inhibition of DNA binding due to direct protein-protein interaction (1990) Cell, 62, pp. 1205-1215