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Phosphorylation of human estrogen receptor α at serine 118 by two distinct signal transduction pathways revealed by phosphorylation-specific antisera

Oncogene volume 21, pages 4921–4931 (2002)Cite this article

Abstract

Estrogen receptor α (ERα) is a transcription factor that regulates expression of target genes in a ligand-dependent manner. Activation of gene expression is mediated by two transcription activation functions AF-1 and AF-2, which act in a promoter- and cell-specific manner. Whilst AF-2 activity is regulated by estrogen (E2) binding, the activity of AF-1 is additionally modulated by phosphorylation at several sites. One of these phosphorylation sites, serine 118 (S118) is of particular interest as its mutation significantly reduces ERα activity. Previous studies have shown that S118 can be phosphorylated by the ERK1/2 mitogen activated protein kinases (MAPK) and by the cyclin-dependent protein kinase Cdk7. In this study we use antisera that specifically recognize ERα phosphorylated at S118 to demonstrate that MAPK phosphorylates S118 in a ligand-independent manner, whereas Cdk7 mediates E2-induced phosphorylation of S118. E2 stimulation of S118 phosphorylation was observed within 10 min of its addition and was maximal at 10−7 M E2. S118 phosphorylation was maximal at 30 min but then declined, such that by 180 min following E2 addition little S118 phosphorylation was evident. S118 phosphorylation was also induced by the partial estrogen antagonist 4-hydroxytamoxifen, but not by the complete antagonist ICI 182, 780. S118 phosphorylation upon addition of the MAPK inducers EGF or PMA followed the expected time courses. Finally, we show that ERα is phosphorylated at S118 in vivo using immunoblotting of extracts prepared from a series of ERα-positive breast tumours.

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References

  • Alessi DR, Saito Y, Campbell DG, Cohen P, Sithanandam G, Rapp U, Ashworth A, Marshall CJ, Cowley S . 1994 EMBO J. 13: 1610–1619

  • Alessi DR, Smythe C, Keyse SM . 1993 Oncogene 8: 2015–2020

  • Ali S, Coombes RC . 2000 J. Mamm. Gland Biol. Neoplasia 5: 271–281

  • Ali S, Lutz Y, Bellocq J-P, Chenard-Neu M-P, Rouyer N, Metzger D . 1993a Hybridoma 12: 391–405

  • Ali S, Metzger D, Bornert JM, Chambon P . 1993b EMBO J. 12: 1153–1160

  • Arnold SF, Obourn JD, Jaffe H, Notides AC . 1994 Mol. Endocrinol. 8: 1208–1214

  • Arnold SF, Obourn JD, Jaffe H, Notides AC . 1995 J. Steroid Biochem. Mol. Biol. 55: 163–172

  • Barettino D, Vivanco Ruiz MM, Stunnenberg HG . 1994 EMBO J. 13: 3039–3049

  • Beato M, Herrlich P, Schutz G . 1995 Cell 83: 851–857

  • Berry M, Metzger D, Chambon P . 1990 EMBO J. 9: 2811–2818

  • Berthois Y, Katzenellenbogen JA, Katzenellenbogen BS . 1986 Proc. Natl. Acad. Sci. USA 83: 2496–2500

  • Blumberg B, Evans RM . 1998 Genes Dev. 12: 3149–3155

  • Brzozowski AM, Pike AC, Dauter Z, Hubbard RE, Bonn T, Engstrom O, Ohman L, Greene GL, Gustafsson JA, Carlquist M . 1997 Nature 389: 753–758

  • Bunone G, Briand PA, Miksicek RJ, Picard D . 1996 EMBO J. 15: 2174–2183

  • Chen D, Riedl T, Washbrook E, Pace PE, Coombes RC, Egly JM, Ali S . 2000 Mol. Cell 6: 127–137

  • Couse, Korach KS . 1999 Endocr. Rev. 20: 358–417

  • Coutts AS, Murphy LC . 1998 Cancer Res. 58: 4071–4074

  • Danielian PS, White R, Lees JA, Parker MG . 1992 EMBO J. 11: 1025–1033

  • Durand B, Saunders M, Gaudon C, Roy B, Losson R, Chambon P . 1994 EMBO J. 13: 5370–5382

  • Gee JM, Robertson JF, Ellis IO, Nicholson RI . 2001 Int. J. Cancer 95: 247–254

  • Glass CK, Rosenfeld MG . 2000 Genes Dev. 14: 121–141

  • Gonzalez FA, Raden DL, Davis RJ . 1991 J. Biol. Chem. 266: 22159–22163

  • Green S, Walter P, Kumar V, Krust A, Bornert JM, Argos P, Chambon P . 1986 Nature 320: 134–139

  • Gronemeyer H . 1991 Annu. Rev. Genet. 25: 89–123

  • Horwitz KB . 1994 J. Steroid Biochem. Mol. Biol. 49: 295–302

  • Horwitz KB, Koseki Y, McGuire WL . 1978 Endocrinology 103: 1742–1751

  • Howe LR, Leevers SJ, Gomez N, Nakielny S, Cohen P, Marshall CJ . 1992 Cell 71: 335–342

  • Joel PB, Traish AM, Lannigan DA . 1998 J. Biol. Chem. 273: 13317–13323

  • Kato S, Endoh H, Masuhiro Y, Kitamoto T, Uchiyama S, Sasaki H, Masushige S, Gotoh Y, Nishida E, Kawashima H, Metzger D, Chambon P . 1995 Science 270: 1491–1494

  • Keyse SM, Emslie EA . 1992 Nature 359: 644–647

  • Kobayashi Y, Kitamoto T, Masuhiro Y, Watanabe M, Kase T, Metzger D, Yanagisawa J, Kato S . 2000 J. Biol. Chem. 275: 15645–15651

  • Le Goff P, Montano MM, Schodin DJ, Katzenellenbogen BS . 1994 J. Biol. Chem. 269: 4458–4466

  • Lees JA, Fawell SE, Parker MG . 1989 Nucleic Acids Res. 17: 5477–5488

  • Makishima M, Okamoto AY, Repa JJ, Tu H, Learned RM, Luk A, Hull MV, Lustig KD, Mangelsdorf DJ, Shan B . 1999 Science 284: 1362–1365

  • Mangelsdorf DJ, Thummel C, Beato M, Herrlich P, Schutz G, Umesono K, Blumberg B, Kastner P, Mark M, Chambon P, Evans RM . 1995 Cell 83: 835–839

  • McDonnell DP, Clemm DL, Hermann T, Goldman ME, Pike JW . 1995 Mol. Endocrinol. 9: 659–669

  • McGuire WL . 1978 Semin. Oncol. 5: 428–433

  • Metzger D, Ali S, Bornert JM, Chambon P . 1995a J. Biol. Chem. 270: 9535–9542

  • Metzger D, Berry M, Ali S, Chambon P . 1995b Mol. Endocrinol. 9: 579–591

  • Moras D, Gronemeyer H . 1998 Curr. Opin. Cell. Biol. 10: 384–391

  • Parks DJ, Blanchard SG, Bledsoe RK, Chandra G, Consler TG, Kliewer SA, Stimmel JB, Willson TM, Zavacki AM, Moore DD, Lehmann JM . 1999 Science 284: 1365–1368

  • Robertson JF . 1996 Br. J. Cancer 73: 5–12

  • Robinson MJ, Stippec SA, Goldsmith E, White MA, Cobb MH . 1998 Curr. Biol. 8: 1141–1150

  • Rogatsky I, Trowbridge JM, Garabedian MJ . 1999 J. Biol. Chem. 274: 22296–22302

  • Saatcioglu F, Bartunek P, Deng T, Zenke M, Karin M . 1993 Mol. Cell. Biol. 13: 3675–3685

  • Shao D, Lazar MA . 1999 J. Clin. Invest. 103: 1617–1618

  • Shiau AK, Barstad D, Loria PM, Cheng L, Kushner PJ, Agard DA, Greene GL . 1998 Cell 95: 927–937

  • Sivaraman VS, Wang H, Nuovo GJ, Malbon CC . 1997 J. Clin. Invest. 99: 1478–1483

  • Smith CL . 1998 Biol. Reprod. 58: 627–632

  • Tassan JP, Schultz SJ, Bartek J, Nigg EA . 1994 J. Cell. Biol. 127: 467–478

  • Tora L, Mullick A, Metzger D, Ponglikitmongkol M, Park I, Chambon P . 1989a EMBO J. 8: 1981–1986

  • Tora L, White J, Brou C, Tasset D, Webster N, Scheer E, Chambon P . 1989b Cell 59: 477–487

  • Tsai MJ, O'Malley BW . 1994 Annu. Rev. Biochem. 63: 451–486

  • Wang H, Chen J, Hollister K, Sowers LC, Forman BM . 1999 Mol. Cell. 3: 543–553

  • Webster NJ, Green S, Jin JR, Chambon P . 1988 Cell 54: 199–207

  • Yankulov K, Yamashita K, Roy R, Egly JM, Bentley DL . 1995 J. Biol. Chem. 270: 23922–23925

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Acknowledgements

We are grateful to Prof Pierre Chambon and Dr Daniel Metzger for the ER plasmids and for expression vectors. We would also like to thank Prof Chris Marshall for providing purified ERK2 protein and plasmids and for helpful discussions. Our thanks also go to Drs S Keyse, E Nigg, T Makela, DO Morgan, G Peters and MH Cobb. For technical help we thank Xiaomei Ouyang. This work was supported by the Cancer Research UK and the Mandeville Trust.

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Author notes

  1. Paul E Pace

    Present address: PNAC, MRC Laboratory of Molecular Biology, Hills Road, Cambridge, CB2 2QH, UK

  2. Richard J Epstein

    Present address: Division of Medical Sciences, National Cancer Centre, Singapore

Authors and Affiliations

  1. Department of Cancer Medicine, Imperial College of Science, Technology and Medicine, Hammersmith Hospital Campus, Du Cane Road, London, W12 0NN, UK

    Dongsheng Chen, Elinor Washbrook, Naveed Sarwar, Paul E Pace, Vatsala Thirunuvakkarasu, Jacqueline Taylor, Richard J Epstein, R Charles Coombes & Simak Ali

  2. Department of Molecular & Cellular Pathology, University of Dundee, Ninewells Hospital & Medical School, Dundee, DD1 9SY, UK

    Gaynor J Bates & Frances V Fuller-Pace

  3. Institut de Genetique et de Biologie Moleculaire et Cellulaire, Université Louis Pasteur, BP 163, Illkirch Cedex Strasbourg, F-67404, France

    Jean-Marc Egly

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Correspondence to Simak Ali.

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Chen, D., Washbrook, E., Sarwar, N. et al. Phosphorylation of human estrogen receptor α at serine 118 by two distinct signal transduction pathways revealed by phosphorylation-specific antisera. Oncogene 21, 4921–4931 (2002). https://doi.org/10.1038/sj.onc.1205420

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