Home LiteratureArticle Details
PMID: 10655479 Published · ppublish English Journal Article Research Support, U.S. Gov't, P.H.S.

Phosphorylation of human progesterone receptors at serine-294 by mitogen-activated protein kinase signals their degradation by the 26S proteasome.

Proceedings of the National Academy of Sciences of the United States of America ·Vol. 97 ·No. 3 ·2000-02-01 ·Pages 1032-7

Lange CA, Shen T, Horwitz KB

Abstract

Ligand-dependent down-regulation that leads to rapid and extensive loss of protein is characteristic of several nuclear steroid receptors, including human progesterone receptors (PRs). In breast cancer cells, >95% of PRs are degraded 6 h after the start of progestin treatment. The mechanism for down-regulation is unknown. We examined the role of PR phosphorylation by mitogen-activated protein kinases (MAPKs) in this process. Lactacystin and calpain inhibitor I, specific inhibitors of the 26S proteasome, blocked progestin-induced down-regulation, and ubiquitinated conjugates of PR accumulated in cells. Ligand-dependent PR degradation was also blocked by specific inhibition of p42 and p44 MAPKs. To define the targets of phosphorylation by this kinase, two serine/proline MAPK consensus sites on PR were mutated. We demonstrate that mutation of PR serine-294 to alanine (S294A) specifically and completely prevents ligand-dependent receptor down-regulation. We also find that rapid, ligand-independent degradation of immature PR intermediates occurs by a proteasome-mediated pathway. These results demonstrate that PR destruction, by either of two alternate routes, is mediated by the 26S proteasome. Specifically, down-regulation of mature PRs occurs by a mechanism in which ligand binding activates PR phosphorylation by MAPKs at a unique serine residue, which then targets the receptors for degradation.

MeSH Terms
Acetylcysteine/analogs & derivatives,pharmacology Breast Neoplasms/chemistry Consensus Sequence Cysteine Endopeptidases/drug effects,metabolism Down-Regulation/physiology Enzyme Inhibitors/pharmacology Female Glycoproteins/pharmacology HeLa Cells/metabolism Humans Imidazoles/pharmacology MAP Kinase Signaling System Mitogen-Activated Protein Kinase 1/antagonists & inhibitors,metabolism Mitogen-Activated Protein Kinase 3 Mitogen-Activated Protein Kinases/antagonists & inhibitors,metabolism Multienzyme Complexes/drug effects,metabolism Neoplasm Proteins/metabolism Phosphorylation Phosphoserine/chemistry Promegestone/pharmacology Proteasome Endopeptidase Complex Protein Isoforms/metabolism Protein Processing, Post-Translational Pyridines/pharmacology Receptors, Progesterone/chemistry,metabolism Recombinant Fusion Proteins/metabolism Ubiquitins/metabolism
Chemicals
Enzyme Inhibitors Glycoproteins Imidazoles Multienzyme Complexes Neoplasm Proteins Protein Isoforms Pyridines Receptors, Progesterone Recombinant Fusion Proteins Ubiquitins calpain inhibitors lactacystin Phosphoserine Promegestone Mitogen-Activated Protein Kinase 1 Mitogen-Activated Protein Kinase 3 Mitogen-Activated Protein Kinases Cysteine Endopeptidases Proteasome Endopeptidase Complex SB 203580 4-(4-fluorophenyl)-2-(4-hydroxyphenyl)-5-(4-pyridyl)imidazole Acetylcysteine
Authors & Affiliations
3 authors, click to expand affiliations / ORCID
Lange C A
Department of Medicine, The Molecular Biology Program, and The Colorado Cancer Center, University of Colorado Health Sciences Center, Denver, CO 80262, USA.
Shen T
Horwitz K B
References (50)
50 references, click to expand
  1. Signal-induced site-specific phosphorylation targets I kappa B alpha to the ubiquitin-proteasome pathway.
    Genes Dev. 1995 Jul 1;9(13):1586-97 PMID: 7628694
  2. Convergence of progesterone with growth factor and cytokine signaling in breast cancer. Progesterone receptors regulate signal transducers and activators of transcription expression and activity.
    J Biol Chem. 1998 Nov 20;273(47):31317-26 PMID: 9813040
  3. Role of phosphorylation on DNA binding and transcriptional functions of human progesterone receptors.
    J Biol Chem. 1996 Jun 7;271(23):13308-16 PMID: 8662865
  4. Inhibition of proteasome activities and subunit-specific amino-terminal threonine modification by lactacystin.
    Science. 1995 May 5;268(5211):726-31 PMID: 7732382
  5. Phosphotryptic peptide analysis of human progesterone receptor. New phosphorylated sites formed in nuclei after hormone treatment.
    J Biol Chem. 1989 Apr 15;264(11):6520-8 PMID: 2703504
  6. Steroid hormone receptors and their regulation by phosphorylation.
    Biochem J. 1996 Nov 1;319 ( Pt 3):657-67 PMID: 8920964
  7. The mitogen-activated protein kinase signal transduction pathway.
    J Biol Chem. 1993 Jul 15;268(20):14553-6 PMID: 8325833
  8. Mutagenic analysis of the destruction signal of mitotic cyclins and structural characterization of ubiquitinated intermediates.
    Mol Biol Cell. 1996 Sep;7(9):1343-57 PMID: 8885231
  9. Progesterone receptor regulation in T47D human breast cancer cells: analysis by density labeling of progesterone receptor synthesis and degradation and their modulation by progestin.
    Endocrinology. 1988 Apr;122(4):1532-40 PMID: 3345726
  10. Tyrosine kinase/p21ras/MAP-kinase pathway activation by estradiol-receptor complex in MCF-7 cells.
    EMBO J. 1996 Mar 15;15(6):1292-300 PMID: 8635462
  11. Progesterone receptor phosphorylation complexities in defining a functional role.
    Trends Endocrinol Metab. 1993 Jan-Feb;4(1):1-7 PMID: 18407126
  12. Ornithine decarboxylase is degraded by the 26S proteasome without ubiquitination.
    Nature. 1992 Dec 10;360(6404):597-9 PMID: 1334232
  13. Disruption of the Raf-1-Hsp90 molecular complex results in destabilization of Raf-1 and loss of Raf-1-Ras association.
    J Biol Chem. 1995 Oct 13;270(41):24585-8 PMID: 7592678
  14. Antagonist-occupied human progesterone receptors bound to DNA are functionally switched to transcriptional agonists by cAMP.
    J Biol Chem. 1993 May 5;268(13):9262-6 PMID: 8387487
  15. Proteasome-dependent degradation of the human estrogen receptor.
    Proc Natl Acad Sci U S A. 1999 Mar 2;96(5):1858-62 PMID: 10051559
  16. Turnover of cyclin E by the ubiquitin-proteasome pathway is regulated by cdk2 binding and cyclin phosphorylation.
    Genes Dev. 1996 Aug 15;10(16):1979-90 PMID: 8769642
  17. Phosphorylation of human progesterone receptor by cyclin-dependent kinase 2 on three sites that are authentic basal phosphorylation sites in vivo.
    Mol Endocrinol. 1997 Jun;11(6):823-32 PMID: 9171245
  18. Progesterone receptor structure and function altered by geldanamycin, an hsp90-binding agent.
    Mol Cell Biol. 1995 Dec;15(12):6804-12 PMID: 8524246
  19. The proteasome is involved in angiogenesis.
    Biochem Biophys Res Commun. 1998 May 8;246(1):243-8 PMID: 9600100
  20. In situ photolinked nuclear progesterone receptors of human breast cancer cells: subunit molecular weights after transformation and translocation.
    Endocrinology. 1983 Dec;113(6):2195-201 PMID: 6685620
  21. Cyclin is degraded by the ubiquitin pathway.
    Nature. 1991 Jan 10;349(6305):132-8 PMID: 1846030
  22. Ubiquitin-dependent c-Jun degradation in vivo is mediated by the delta domain.
    Cell. 1994 Sep 9;78(5):787-98 PMID: 8087846
  23. p34Cdc28-mediated control of Cln3 cyclin degradation.
    Mol Cell Biol. 1995 Feb;15(2):731-41 PMID: 7823941
  24. Roles of ubiquitin-mediated proteolysis in cell cycle control.
    Curr Opin Cell Biol. 1997 Dec;9(6):788-99 PMID: 9425343
  25. Mouse glucocorticoid receptor phosphorylation status influences multiple functions of the receptor protein.
    J Biol Chem. 1997 Apr 4;272(14):9287-93 PMID: 9083064
  26. Convergence of progesterone and epidermal growth factor signaling in breast cancer. Potentiation of mitogen-activated protein kinase pathways.
    J Biol Chem. 1998 Nov 20;273(47):31308-16 PMID: 9813039
  27. Targeting of substrates to the 26S proteasome.
    FASEB J. 1997 Nov;11(13):1055-66 PMID: 9367341
  28. New T47D breast cancer cell lines for the independent study of progesterone B- and A-receptors: only antiprogestin-occupied B-receptors are switched to transcriptional agonists by cAMP.
    Cancer Res. 1994 Jul 15;54(14):3868-77 PMID: 8033109
  29. Inhibition of cyclin D1 phosphorylation on threonine-286 prevents its rapid degradation via the ubiquitin-proteasome pathway.
    Genes Dev. 1997 Apr 15;11(8):957-72 PMID: 9136925
  30. A protein kinase involved in the regulation of inflammatory cytokine biosynthesis.
    Nature. 1994 Dec 22-29;372(6508):739-46 PMID: 7997261
  31. Lactacystin, proteasome function, and cell fate.
    J Biol Chem. 1998 Apr 10;273(15):8545-8 PMID: 9535824
  32. Variant T47D human breast cancer cells with high progesterone-receptor levels despite estrogen and antiestrogen resistance.
    Cell. 1982 Mar;28(3):633-42 PMID: 7200400
  33. A synthetic inhibitor of the mitogen-activated protein kinase cascade.
    Proc Natl Acad Sci U S A. 1995 Aug 15;92(17):7686-9 PMID: 7644477
  34. Evidence for enhanced ubiquitin-mediated proteolysis of the chicken progesterone receptor by progesterone.
    Life Sci. 1998;63(17):1505-12 PMID: 9808061
  35. Pathways of ubiquitin conjugation.
    FASEB J. 1997 Dec;11(14):1257-68 PMID: 9409544
  36. Specific progesterone receptors in human breast cancer.
    Steroids. 1975 Apr;25(4):497-505 PMID: 165596
  37. Prognostic factors in breast cancer.
    Semin Oncol. 1992 Jun;19(3):244-53 PMID: 1351692
  38. Cell cycle regulation by the ubiquitin pathway.
    FASEB J. 1997 Nov;11(13):1067-75 PMID: 9367342
  39. Multiple human progesterone receptor messenger ribonucleic acids and their autoregulation by progestin agonists and antagonists in breast cancer cells.
    Mol Endocrinol. 1988 Jan;2(1):62-72 PMID: 3398843
  40. Endoproteolytic cleavage and proteasomal degradation of presenilin 2 in transfected cells.
    J Biol Chem. 1997 Apr 25;272(17):11006-10 PMID: 9110991
  41. Polyubiquitination and proteasomal degradation of the p185c-erbB-2 receptor protein-tyrosine kinase induced by geldanamycin.
    J Biol Chem. 1996 Sep 13;271(37):22796-801 PMID: 8798456
  42. Activation of the Src/p21ras/Erk pathway by progesterone receptor via cross-talk with estrogen receptor.
    EMBO J. 1998 Apr 1;17(7):2008-18 PMID: 9524123
  43. Activation of cyclin E/CDK2 is coupled to site-specific autophosphorylation and ubiquitin-dependent degradation of cyclin E.
    EMBO J. 1996 Aug 15;15(16):4182-93 PMID: 8861947
  44. Stable and specific binding of heat shock protein 90 by geldanamycin disrupts glucocorticoid receptor function in intact cells.
    Mol Endocrinol. 1996 Jun;10(6):705-12 PMID: 8776730
  45. The hsp90-based chaperone system: involvement in signal transduction from a variety of hormone and growth factor receptors.
    Proc Soc Exp Biol Med. 1998 Apr;217(4):420-34 PMID: 9521088
  46. Estrogen-induced activation of mitogen-activated protein kinase requires mobilization of intracellular calcium.
    Proc Natl Acad Sci U S A. 1999 Apr 13;96(8):4686-91 PMID: 10200323
  47. Identification of a group of Ser-Pro motif hormone-inducible phosphorylation sites in the human progesterone receptor.
    Mol Endocrinol. 1995 Aug;9(8):1029-40 PMID: 7476977
  48. Enhanced in vitro cytotoxicity and cytostasis of the combination of onconase with a proteasome inhibitor.
    Int J Oncol. 1998 Oct;13(4):633-44 PMID: 9735389
  49. Cellular progesterone receptor phosphorylation in response to ligands activating protein kinases.
    Biochem Biophys Res Commun. 1987 Aug 14;146(3):1357-65 PMID: 3497636
  50. The ubiquitin-proteasome pathway in cancer.
    Br J Cancer. 1998;77(3):448-55 PMID: 9472642
Article Info
Journal
Proceedings of the National Academy of Sciences of the United States of America
Abbr.
Proc Natl Acad Sci U S A
ISSN
0027-8424
DOI
10.1073/pnas.97.3.1032
Published
2000-02-01
Pages
1032-7
Language
English
Region
United States
NLM ID
7505876
PMCID
PMC15511
Subset
IM
Grants
NIDDK NIH HHS · DK53825 · United States
NIDDK NIH HHS · DK48238 · United States
NCI NIH HHS · R01 CA026869 · United States
NCI NIH HHS · R37 CA026869 · United States
NIDDK NIH HHS · R01 DK053825 · United States
NCI NIH HHS · CA26869 · United States
External Links
PubMed source DOI: 10.1073/pnas.97.3.1032 PMC Full Text View journal details
Analysis Services
Analysis Services

Contact

No. 2 Wenbo Road, Zhangqiu District, Jinan, Shandong

Qilu Normal University · Genelibs Bioinformatics Lab

750 Shunhua Rd, Jinan

2F, Bldg F, University Science Park

Tel: 0531-88819269

WeChat Official Account

Follow our WeChat subscription account for real-time updates and the latest in medical and biological research.


Business Email

E-mail: product@genelibs.com

Copyright © 2026.Genelibs Jinan Honghui Biotechnology Co., Ltd. 鲁ICP备20007159号-2