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PMID: 15743833 Published · ppublish English Journal Article

Ligand-specific dynamics of the progesterone receptor in living cells and during chromatin remodeling in vitro.

Molecular and cellular biology ·Vol. 25 ·No. 6 ·2005-03-00 ·Pages 2406-18

Rayasam GV, Elbi C, Walker DA, Wolford R, Fletcher TM, Edwards DP, Hager GL

Abstract

Progesterone receptor (PR), a member of the nuclear receptor superfamily, is a key regulator of several processes in reproductive function. We have studied the dynamics of the interaction of PR with a natural target promoter in living cells through the use of fluorescence recovery after photobleaching (FRAP) analysis and also have characterized the dynamics of the interaction of PR with the mouse mammary tumor virus (MMTV) promoter reconstituted into chromatin in vitro. In photobleaching experiments, PR in the presence of the agonist R5020 exhibits rapid exchange with the MMTV promoter in living cells. Two PR antagonists, RU486 and ZK98299, have opposite effects on receptor dynamics in vivo. In the presence of RU486, PR binds to the promoter and is exchanged more slowly than the agonist-activated receptor. In contrast, PR bound to ZK98299 is not localized to the promoter and exhibits higher mobility in the nucleoplasm than the agonist-bound receptor. Significantly, PR bound to R5020 or RU486 can recruit the SWI/SNF chromatin remodeling complex to the promoter, but PR activated with ZK98299 cannot. Furthermore, we found ligand-specific active displacement of PR from the MMTV promoter during chromatin remodeling in vitro and conclude that the interaction of PR with chromatin is highly dynamic both in vivo and in vitro. We propose that factor displacement during chromatin remodeling is an important component of receptor mobility and that ligand-specific interactions with remodeling complexes can strongly influence receptor nuclear dynamics and rates of exchange with chromatin in living cells.

MeSH Terms
Cell Nucleus/metabolism Cells, Cultured Chromatin/metabolism Chromatin Assembly and Disassembly/genetics,physiology DNA Helicases Fluorescence Recovery After Photobleaching Gonanes/pharmacology Green Fluorescent Proteins/analysis,genetics Humans Ligands Mammary Tumor Virus, Mouse/genetics Mifepristone/pharmacology Nuclear Proteins/genetics,metabolism Promegestone/pharmacology Promoter Regions, Genetic/genetics Receptors, Progesterone/drug effects,genetics,metabolism Transcription Factors/genetics,metabolism
Chemicals
Chromatin Gonanes Ligands Nuclear Proteins Receptors, Progesterone Transcription Factors enhanced green fluorescent protein Green Fluorescent Proteins Mifepristone Promegestone SMARCA4 protein, human Smarca4 protein, mouse DNA Helicases onapristone
Authors & Affiliations
7 authors, click to expand affiliations / ORCID
Rayasam Geetha V
Laboratory of Receptor Biology and Gene Expression, Building 41, B602, National Cancer Institute, National Institutes of Health, 41 Library Dr., Bethesda, MD 20892-5055, USA.
Elbi Cem
Walker Dawn A
Wolford Ronald
Fletcher Terace M
Edwards Dean P
Hager Gordon L
References (46)
46 references, click to expand
  1. Using inducible vectors to study intracellular trafficking of GFP-tagged steroid/nuclear receptors in living cells.
    Methods. 1999 Nov;19(3):386-93 PMID: 10579933
  2. The activity of mammalian brm/SNF2alpha is dependent on a high-mobility-group protein I/Y-like DNA binding domain.
    Mol Cell Biol. 1999 Jun;19(6):3931-9 PMID: 10330133
  3. Mammalian SWI-SNF complexes contribute to activation of the hsp70 gene.
    Mol Cell Biol. 2000 Apr;20(8):2839-51 PMID: 10733587
  4. The opposing transcriptional activities of the two isoforms of the human progesterone receptor are due to differential cofactor binding.
    Mol Cell Biol. 2000 May;20(9):3102-15 PMID: 10757795
  5. Structure and dynamic properties of a glucocorticoid receptor-induced chromatin transition.
    Mol Cell Biol. 2000 Sep;20(17):6466-75 PMID: 10938123
  6. Understanding nuclear receptor function: from DNA to chromatin to the interphase nucleus.
    Prog Nucleic Acid Res Mol Biol. 2001;66:279-305 PMID: 11051767
  7. Protein dynamics: implications for nuclear architecture and gene expression.
    Science. 2001 Feb 2;291(5505):843-7 PMID: 11225636
  8. Ligand-mediated assembly and real-time cellular dynamics of estrogen receptor alpha-coactivator complexes in living cells.
    Mol Cell Biol. 2001 Jul;21(13):4404-12 PMID: 11390668
  9. Nuclear receptors coordinate the activities of chromatin remodeling complexes and coactivators to facilitate initiation of transcription.
    Oncogene. 2001 May 28;20(24):3047-54 PMID: 11420720
  10. Combinatorial control of gene expression by nuclear receptors and coregulators.
    Cell. 2002 Feb 22;108(4):465-74 PMID: 11909518
  11. ATP-dependent mobilization of the glucocorticoid receptor during chromatin remodeling.
    Mol Cell Biol. 2002 May;22(10):3255-63 PMID: 11971959
  12. Coactivator/corepressor ratios modulate PR-mediated transcription by the selective receptor modulator RU486.
    Proc Natl Acad Sci U S A. 2002 Jun 11;99(12):7940-4 PMID: 12048256
  13. Disassembly of transcriptional regulatory complexes by molecular chaperones.
    Science. 2002 Jun 21;296(5576):2232-5 PMID: 12077419
  14. Dynamic behavior of transcription factors on a natural promoter in living cells.
    EMBO Rep. 2002 Dec;3(12):1188-94 PMID: 12446572
  15. Mechanism of action of progesterone antagonists.
    Exp Biol Med (Maywood). 2002 Dec;227(11):969-80 PMID: 12486206
  16. Cyclic, proteasome-mediated turnover of unliganded and liganded ERalpha on responsive promoters is an integral feature of estrogen signaling.
    Mol Cell. 2003 Mar;11(3):695-707 PMID: 12667452
  17. Differential requirement of SWI/SNF for androgen receptor activity.
    J Biol Chem. 2003 Aug 15;278(33):30605-13 PMID: 12775722
  18. Nuclear receptors: a rendezvous for chromatin remodeling factors.
    Cell. 2003 Aug 8;114(3):277-80 PMID: 12914692
  19. Unfolding the action of progesterone receptors.
    J Biol Chem. 2003 Oct 10;278(41):39261-4 PMID: 12893816
  20. Estrogen receptor-alpha directs ordered, cyclical, and combinatorial recruitment of cofactors on a natural target promoter.
    Cell. 2003 Dec 12;115(6):751-63 PMID: 14675539
  21. Molecular chaperones function as steroid receptor nuclear mobility factors.
    Proc Natl Acad Sci U S A. 2004 Mar 2;101(9):2876-81 PMID: 14978266
  22. Dynamics of nuclear receptor movement and transcription.
    Biochim Biophys Acta. 2004 Mar 15;1677(1-3):46-51 PMID: 15020044
  23. Rapid glucocorticoid receptor exchange at a promoter is coupled to transcription and regulated by chaperones and proteasomes.
    Mol Cell Biol. 2004 Apr;24(7):2682-97 PMID: 15024059
  24. Transcription factors and nuclear receptors interact with the SWI/SNF complex through the BAF60c subunit.
    J Biol Chem. 2004 Apr 16;279(16):16677-86 PMID: 14701856
  25. Rapid periodic binding and displacement of the glucocorticoid receptor during chromatin remodeling.
    Mol Cell. 2004 Apr 23;14(2):163-74 PMID: 15099516
  26. Temporal recruitment of transcription factors and SWI/SNF chromatin-remodeling enzymes during adipogenic induction of the peroxisome proliferator-activated receptor gamma nuclear hormone receptor.
    Mol Cell Biol. 2004 Jun;24(11):4651-63 PMID: 15143161
  27. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding.
    Anal Biochem. 1976 May 7;72:248-54 PMID: 942051
  28. Specific glucocorticoid receptor binding to DNA reconstituted in a nucleosome.
    EMBO J. 1988 Oct;7(10):3073-9 PMID: 2846275
  29. Two types of antiprogestins identified by their differential action in transcriptionally active extracts from T47D cells.
    Nucleic Acids Res. 1991 Mar 25;19(6):1227-34 PMID: 2030942
  30. Effects of the steroid antagonist RU486 on dimerization of the human progesterone receptor.
    Biochemistry. 1992 Nov 3;31(43):10491-501 PMID: 1420166
  31. Roles of SWI1, SWI2, and SWI3 proteins for transcriptional enhancement by steroid receptors.
    Science. 1992 Dec 4;258(5088):1598-604 PMID: 1360703
  32. In vivo evidence against the existence of antiprogestins disrupting receptor binding to DNA.
    Proc Natl Acad Sci U S A. 1993 May 15;90(10):4421-5 PMID: 8506282
  33. A human homologue of Saccharomyces cerevisiae SNF2/SWI2 and Drosophila brm genes potentiates transcriptional activation by the glucocorticoid receptor.
    EMBO J. 1993 Nov;12(11):4279-90 PMID: 8223438
  34. Antiprogestins prevent progesterone receptor binding to hormone responsive elements in vivo.
    Proc Natl Acad Sci U S A. 1994 Nov 22;91(24):11333-7 PMID: 7972059
  35. Dissection of progesterone receptor-mediated chromatin remodeling and transcriptional activation in vivo.
    Genes Dev. 1995 Jun 1;9(11):1366-76 PMID: 7797076
  36. Progesterone receptor and the mechanism of action of progesterone antagonists.
    J Steroid Biochem Mol Biol. 1995 Jun;53(1-6):449-58 PMID: 7626494
  37. Nucleosome positioning on the MMTV LTR results from the frequency-biased occupancy of multiple frames.
    Genes Dev. 1995 Aug 1;9(15):1933-47 PMID: 7649476
  38. The affinity of nuclear factor 1 for its DNA site is drastically reduced by nucleosome organization irrespective of its rotational or translational position.
    J Biol Chem. 1996 Jan 5;271(1):153-9 PMID: 8550551
  39. Ligand-dependent interaction between the estrogen receptor and the human homologues of SWI2/SNF2.
    Gene. 1997 Mar 25;188(1):95-100 PMID: 9099865
  40. The antagonists RU486 and ZK98299 stimulate progesterone receptor binding to deoxyribonucleic acid in vitro and in vivo, but have distinct effects on receptor conformation.
    Endocrinology. 1998 Apr;139(4):1905-19 PMID: 9528977
  41. Chromatin remodelling by the glucocorticoid receptor requires the BRG1 complex.
    Nature. 1998 May 7;393(6680):88-91 PMID: 9590696
  42. The position and length of the steroid-dependent hypersensitive region in the mouse mammary tumor virus long terminal repeat are invariant despite multiple nucleosome B frames.
    Mol Cell Biol. 1998 Jun;18(6):3633-44 PMID: 9584204
  43. High-mobility group chromatin proteins 1 and 2 functionally interact with steroid hormone receptors to enhance their DNA binding in vitro and transcriptional activity in mammalian cells.
    Mol Cell Biol. 1998 Aug;18(8):4471-87 PMID: 9671457
  44. Assembly of MMTV promoter minichromosomes with positioned nucleosomes precludes NF1 access but not restriction enzyme cleavage.
    Nucleic Acids Res. 1998 Aug 15;26(16):3657-66 PMID: 9685480
  45. Differential localization and activity of the A- and B-forms of the human progesterone receptor using green fluorescent protein chimeras.
    Mol Endocrinol. 1999 Mar;13(3):366-75 PMID: 10076994
  46. The glucocorticoid receptor: rapid exchange with regulatory sites in living cells.
    Science. 2000 Feb 18;287(5456):1262-5 PMID: 10678832
Article Info
Journal
Molecular and cellular biology
Abbr.
Mol Cell Biol
ISSN
0270-7306
DOI
10.1128/MCB.25.6.2406-2418.2005
Published
2005-03-00
Pages
2406-18
Language
English
Region
United States
NLM ID
8109087
PMCID
PMC1061598
Subset
IM
External Links
PubMed source DOI: 10.1128/MCB.25.6.2406-2418.2005 PMC Full Text View journal details
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