• We are sorry, but NCBI web applications do not support your browser and may not function properly. More information
Logo of embojLink to Publisher's site
EMBO J. Mar 1993; 12(3): 1153–1160.
PMCID: PMC413317

Modulation of transcriptional activation by ligand-dependent phosphorylation of the human oestrogen receptor A/B region.

Abstract

Using a transient co-transfection system, we show that the human oestrogen receptor (hER) becomes phosphorylated in the presence of oestradiol (E2) as well as in the presence of the anti-oestrogens 4-hydroxy-tamoxifen (OHT) and ICI 164, 384 (ICI), although at lower efficiencies than with E2. There are multiple sites of phosphorylation in hER; using deletion and point mutants one of these sites has been mapped in the N-terminal A/B region at serine 118. Mutation of this serine to alanine caused, in a number of cell types, a significant reduction in transcriptional activation by hER from reporter genes containing an oestrogen response element (ERE), but did not affect the DNA binding properties or nuclear localization of hER. Thus phosphorylation of serine 118 is important for the action of the transcription activation function 1 (AF-1) located in the A/B region of the oestrogen receptor.

Full text

Full text is available as a scanned copy of the original print version. Get a printable copy (PDF file) of the complete article (2.0M), or click on a page image below to browse page by page. Links to PubMed are also available for Selected References.

Selected References

These references are in PubMed. This may not be the complete list of references from this article.
  • Auricchio F. Phosphorylation of steroid receptors. J Steroid Biochem. 1989 Apr;32(4):613–622. [PubMed]
  • Auricchio F, Migliaccio A, Di Domenico M, Nola E. Oestradiol stimulates tyrosine phosphorylation and hormone binding activity of its own receptor in a cell-free system. EMBO J. 1987 Oct;6(10):2923–2929. [PMC free article] [PubMed]
  • Beato M. Gene regulation by steroid hormones. Cell. 1989 Feb 10;56(3):335–344. [PubMed]
  • Berry M, Nunez AM, Chambon P. Estrogen-responsive element of the human pS2 gene is an imperfectly palindromic sequence. Proc Natl Acad Sci U S A. 1989 Feb;86(4):1218–1222. [PMC free article] [PubMed]
  • Berry M, Metzger D, Chambon P. Role of the two activating domains of the oestrogen receptor in the cell-type and promoter-context dependent agonistic activity of the anti-oestrogen 4-hydroxytamoxifen. EMBO J. 1990 Sep;9(9):2811–2818. [PMC free article] [PubMed]
  • Berthois Y, Katzenellenbogen JA, Katzenellenbogen BS. Phenol red in tissue culture media is a weak estrogen: implications concerning the study of estrogen-responsive cells in culture. Proc Natl Acad Sci U S A. 1986 Apr;83(8):2496–2500. [PMC free article] [PubMed]
  • Dauvois S, Danielian PS, White R, Parker MG. Antiestrogen ICI 164,384 reduces cellular estrogen receptor content by increasing its turnover. Proc Natl Acad Sci U S A. 1992 May 1;89(9):4037–4041. [PMC free article] [PubMed]
  • DeFranco DB, Qi M, Borror KC, Garabedian MJ, Brautigan DL. Protein phosphatase types 1 and/or 2A regulate nucleocytoplasmic shuttling of glucocorticoid receptors. Mol Endocrinol. 1991 Sep;5(9):1215–1228. [PubMed]
  • Denton RR, Koszewski NJ, Notides AC. Estrogen receptor phosphorylation. Hormonal dependence and consequence on specific DNA binding. J Biol Chem. 1992 Apr 15;267(11):7263–7268. [PubMed]
  • Evans RM. The steroid and thyroid hormone receptor superfamily. Science. 1988 May 13;240(4854):889–895. [PubMed]
  • Fahrner TJ, Carroll SL, Milbrandt J. The NGFI-B protein, an inducible member of the thyroid/steroid receptor family, is rapidly modified posttranslationally. Mol Cell Biol. 1990 Dec;10(12):6454–6459. [PMC free article] [PubMed]
  • Gaub MP, Rochette-Egly C, Lutz Y, Ali S, Matthes H, Scheuer I, Chambon P. Immunodetection of multiple species of retinoic acid receptor alpha: evidence for phosphorylation. Exp Cell Res. 1992 Aug;201(2):335–346. [PubMed]
  • Glineur C, Zenke M, Beug H, Ghysdael J. Phosphorylation of the v-erbA protein is required for its function as an oncogene. Genes Dev. 1990 Oct;4(10):1663–1676. [PubMed]
  • Green S, Chambon P. Oestradiol induction of a glucocorticoid-responsive gene by a chimaeric receptor. Nature. 1987 Jan 1;325(6099):75–78. [PubMed]
  • Green S, Chambon P. Nuclear receptors enhance our understanding of transcription regulation. Trends Genet. 1988 Nov;4(11):309–314. [PubMed]
  • Green S, Walter P, Kumar V, Krust A, Bornert JM, Argos P, Chambon P. Human oestrogen receptor cDNA: sequence, expression and homology to v-erb-A. Nature. 1986 Mar 13;320(6058):134–139. [PubMed]
  • Gronemeyer H. Transcription activation by estrogen and progesterone receptors. Annu Rev Genet. 1991;25:89–123. [PubMed]
  • Hall FL, Vulliet PR. Proline-directed protein phosphorylation and cell cycle regulation. Curr Opin Cell Biol. 1991 Apr;3(2):176–184. [PubMed]
  • Horwitz KB, McGuire WL. Nuclear mechanisms of estrogen action. Effects of estradiol and anti-estrogens on estrogen receptors and nuclear receptor processing. J Biol Chem. 1978 Nov 25;253(22):8185–8191. [PubMed]
  • Hsieh JC, Jurutka PW, Galligan MA, Terpening CM, Haussler CA, Samuels DS, Shimizu Y, Shimizu N, Haussler MR. Human vitamin D receptor is selectively phosphorylated by protein kinase C on serine 51, a residue crucial to its trans-activation function. Proc Natl Acad Sci U S A. 1991 Oct 15;88(20):9315–9319. [PMC free article] [PubMed]
  • Jackson SP. Regulating transcription factor activity by phosphorylation. Trends Cell Biol. 1992 Apr;2(4):104–108. [PubMed]
  • Kato S, Tora L, Yamauchi J, Masushige S, Bellard M, Chambon P. A far upstream estrogen response element of the ovalbumin gene contains several half-palindromic 5'-TGACC-3' motifs acting synergistically. Cell. 1992 Feb 21;68(4):731–742. [PubMed]
  • Koelle MR, Talbot WS, Segraves WA, Bender MT, Cherbas P, Hogness DS. The Drosophila EcR gene encodes an ecdysone receptor, a new member of the steroid receptor superfamily. Cell. 1991 Oct 4;67(1):59–77. [PubMed]
  • Koike S, Sakai M, Muramatsu M. Molecular cloning and characterization of rat estrogen receptor cDNA. Nucleic Acids Res. 1987 Mar 25;15(6):2499–2513. [PMC free article] [PubMed]
  • Krust A, Green S, Argos P, Kumar V, Walter P, Bornert JM, Chambon P. The chicken oestrogen receptor sequence: homology with v-erbA and the human oestrogen and glucocorticoid receptors. EMBO J. 1986 May;5(5):891–897. [PMC free article] [PubMed]
  • Kumar V, Green S, Staub A, Chambon P. Localisation of the oestradiol-binding and putative DNA-binding domains of the human oestrogen receptor. EMBO J. 1986 Sep;5(9):2231–2236. [PMC free article] [PubMed]
  • Kumar V, Green S, Stack G, Berry M, Jin JR, Chambon P. Functional domains of the human estrogen receptor. Cell. 1987 Dec 24;51(6):941–951. [PubMed]
  • Laudet V, Hänni C, Coll J, Catzeflis F, Stéhelin D. Evolution of the nuclear receptor gene superfamily. EMBO J. 1992 Mar;11(3):1003–1013. [PMC free article] [PubMed]
  • Lees JA, Fawell SE, Parker MG. Identification of two transactivation domains in the mouse oestrogen receptor. Nucleic Acids Res. 1989 Jul 25;17(14):5477–5488. [PMC free article] [PubMed]
  • Metzger D, Losson R, Bornert JM, Lemoine Y, Chambon P. Promoter specificity of the two transcriptional activation functions of the human oestrogen receptor in yeast. Nucleic Acids Res. 1992 Jun 11;20(11):2813–2817. [PMC free article] [PubMed]
  • Migliaccio A, Di Domenico M, Green S, de Falco A, Kajtaniak EL, Blasi F, Chambon P, Auricchio F. Phosphorylation on tyrosine of in vitro synthesized human estrogen receptor activates its hormone binding. Mol Endocrinol. 1989 Jul;3(7):1061–1069. [PubMed]
  • Moreno S, Nurse P. Substrates for p34cdc2: in vivo veritas? Cell. 1990 May 18;61(4):549–551. [PubMed]
  • Norbury CJ, Nurse P. Control of the higher eukaryote cell cycle by p34cdc2 homologues. Biochim Biophys Acta. 1989 Jul 28;989(1):85–95. [PubMed]
  • Nunez AM, Berry M, Imler JL, Chambon P. The 5' flanking region of the pS2 gene contains a complex enhancer region responsive to oestrogens, epidermal growth factor, a tumour promoter (TPA), the c-Ha-ras oncoprotein and the c-jun protein. EMBO J. 1989 Mar;8(3):823–829. [PMC free article] [PubMed]
  • Pakdel F, Le Gac F, Le Goff P, Valotaire Y. Full-length sequence and in vitro expression of rainbow trout estrogen receptor cDNA. Mol Cell Endocrinol. 1990 Jul 9;71(3):195–204. [PubMed]
  • Power RF, Mani SK, Codina J, Conneely OM, O'Malley BW. Dopaminergic and ligand-independent activation of steroid hormone receptors. Science. 1991 Dec 13;254(5038):1636–1639. [PubMed]
  • Ptashne M. How eukaryotic transcriptional activators work. Nature. 1988 Oct 20;335(6192):683–689. [PubMed]
  • Rochette-Egly C, Lutz Y, Saunders M, Scheuer I, Gaub MP, Chambon P. Retinoic acid receptor gamma: specific immunodetection and phosphorylation. J Cell Biol. 1991 Oct;115(2):535–545. [PMC free article] [PubMed]
  • Rochette-Egly C, Gaub MP, Lutz Y, Ali S, Scheuer I, Chambon P. Retinoic acid receptor-beta: immunodetection and phosphorylation on tyrosine residues. Mol Endocrinol. 1992 Dec;6(12):2197–2209. [PubMed]
  • Tasset D, Tora L, Fromental C, Scheer E, Chambon P. Distinct classes of transcriptional activating domains function by different mechanisms. Cell. 1990 Sep 21;62(6):1177–1187. [PubMed]
  • 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. [PubMed]
  • Tora L, Mullick A, Metzger D, Ponglikitmongkol M, Park I, Chambon P. The cloned human oestrogen receptor contains a mutation which alters its hormone binding properties. EMBO J. 1989 Jul;8(7):1981–1986. [PMC free article] [PubMed]
  • van Laar JH, Berrevoets CA, Trapman J, Zegers ND, Brinkmann AO. Hormone-dependent androgen receptor phosphorylation is accompanied by receptor transformation in human lymph node carcinoma of the prostate cells. J Biol Chem. 1991 Feb 25;266(6):3734–3738. [PubMed]
  • Vulliet PR, Hall FL, Mitchell JP, Hardie DG. Identification of a novel proline-directed serine/threonine protein kinase in rat pheochromocytoma. J Biol Chem. 1989 Sep 25;264(27):16292–16298. [PubMed]
  • Washburn T, Hocutt A, Brautigan DL, Korach KS. Uterine estrogen receptor in vivo: phosphorylation of nuclear specific forms on serine residues. Mol Endocrinol. 1991 Feb;5(2):235–242. [PubMed]
  • Webster NJ, Green S, Jin JR, 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. [PubMed]
  • Weigel NL, Carter TH, Schrader WT, O'Malley BW. Chicken progesterone receptor is phosphorylated by a DNA-dependent protein kinase during in vitro transcription assays. Mol Endocrinol. 1992 Jan;6(1):8–14. [PubMed]
  • Weiler IJ, Lew D, Shapiro DJ. The Xenopus laevis estrogen receptor: sequence homology with human and avian receptors and identification of multiple estrogen receptor messenger ribonucleic acids. Mol Endocrinol. 1987 May;1(5):355–362. [PubMed]
  • White R, Lees JA, Needham M, Ham J, Parker M. Structural organization and expression of the mouse estrogen receptor. Mol Endocrinol. 1987 Oct;1(10):735–744. [PubMed]

Articles from The EMBO Journal are provided here courtesy of The European Molecular Biology Organization

Formats:

Related citations in PubMed

See reviews...See all...

Cited by other articles in PMC

See all...

Links

  • Compound
    Compound
    PubChem Compound links
  • MedGen
    MedGen
    Related information in MedGen
  • PubMed
    PubMed
    PubMed citations for these articles
  • Substance
    Substance
    PubChem Substance links

Recent Activity

Your browsing activity is empty.

Activity recording is turned off.

Turn recording back on

See more...