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PMID: 9343435 Published · ppublish English Comparative Study Journal Article

Specific DNA binding of Stat5, but not of glucocorticoid receptor, is required for their functional cooperation in the regulation of gene transcription.

Molecular and cellular biology ·Vol. 17 ·No. 11 ·1997-11-00 ·Pages 6708-16

Stoecklin E, Wissler M, Moriggl R, Groner B

Abstract

Prolactin and glucocorticoid hormone are signals which regulate the transcription of milk protein genes in mammary epithelial cells. We have investigated the molecular mechanisms by which these hormones cooperate in the induction of transcription. Both hormones activate latent transcription factors in the cytoplasm of mammary epithelial cells. Prolactin exerts its effect through binding to the extracellular domain of the prolactin receptor and through receptor dimerization. This leads to the activation of a protein tyrosine kinase (Jak2), which is noncovalently associated with the cytoplasmic domain of the prolactin receptor. Jak2 phosphorylates the signal transducer and transcription activator (Stat5) which causes its dimerization and nuclear translocation where Stat5 specifically binds to sequence elements in the promoter regions of milk protein genes. In comparison, the glucocorticoid receptor is activated by a lipophilic steroid ligand in the cytoplasm which causes allosteric changes in the molecule, dimerization, and nuclear localization. It has been demonstrated that Stat5 and the glucocorticoid receptor form a molecular complex which cooperates in the induction of transcription of the beta-casein gene. We have defined the DNA sequence requirements for this cooperative mechanism and have delimited the functional domains in Stat5 and the glucocorticoid receptor that are necessary for the functional interaction. We find that the Stat5 response element (Stat5RE) within the beta-casein gene promoter is sufficient to elicit the cooperative action of Stat5 and the glucocorticoid receptor on transcription. Activation of Stat5 through phosphorylation of tyrosine 694 is an absolute prerequisite for transcription. Deletion of the transactivation domain of Stat5 results in a molecule which cannot mediate transactivation by itself but can still cooperate with the glucocorticoid receptor. Mutated variants of the glucocorticoid receptor with a nonfunctional DNA binding domain or a DNA binding domain contributed by the estrogen receptor are still able to cooperate with Stat5 in transcriptional induction. Deletion of the ligand binding domain of the glucocorticoid receptor does not impede cooperation with Stat5, whereas deletion of the AF-1 transactivation domain does prevent cooperation. Our results indicate that the glucocorticoid receptor acts as a ligand-dependent coactivator of Stat5 independently of its DNA binding function.

MeSH Terms
Animals Binding Sites COS Cells Caseins/biosynthesis,genetics DNA-Binding Proteins/genetics,metabolism Dexamethasone/pharmacology Gene Expression Regulation Glucocorticoids/pharmacology Ligands Milk Proteins Models, Genetic Phosphorylation Prolactin/pharmacology Promoter Regions, Genetic Protein Binding Receptors, Glucocorticoid/genetics,metabolism Recombinant Fusion Proteins/biosynthesis STAT5 Transcription Factor Sequence Deletion Signal Transduction Trans-Activators/genetics,metabolism Transcription, Genetic Tyrosine/metabolism
Chemicals
Caseins DNA-Binding Proteins Glucocorticoids Ligands Milk Proteins Receptors, Glucocorticoid Recombinant Fusion Proteins STAT5 Transcription Factor Trans-Activators Tyrosine Dexamethasone Prolactin
Authors & Affiliations
4 authors, click to expand affiliations / ORCID
Stoecklin E
Institute for Experimental Cancer Research, Tumor Biology Center Freiburg, Germany.
Wissler M
Moriggl R
Groner B
References (78)
78 references, click to expand
  1. Involvement of the transcription factor PU.1/Spi-1 in myeloid cell-restricted expression of an interferon-inducible gene encoding the human high-affinity Fc gamma receptor.
    Mol Cell Biol. 1994 Aug;14(8):5023-31 PMID: 8035786
  2. Hormonal regulation of transcription factor activity in mammary epithelial cells.
    Mol Cell Endocrinol. 1994 Apr;100(1-2):109-14 PMID: 8056143
  3. A distinct modulating domain in glucocorticoid receptor monomers in the repression of activity of the transcription factor AP-1.
    EMBO J. 1994 Sep 1;13(17):4087-95 PMID: 8076604
  4. Prolactin induces phosphorylation of Tyr694 of Stat5 (MGF), a prerequisite for DNA binding and induction of transcription.
    EMBO J. 1994 Sep 15;13(18):4361-9 PMID: 7925280
  5. A CBP integrator complex mediates transcriptional activation and AP-1 inhibition by nuclear receptors.
    Cell. 1996 May 3;85(3):403-14 PMID: 8616895
  6. Interleukin-2 activation of STAT5 requires the convergent action of tyrosine kinases and a serine/threonine kinase pathway distinct from the Raf1/ERK2 MAP kinase pathway.
    EMBO J. 1996 Apr 15;15(8):1902-13 PMID: 8617237
  7. GRIP1, a novel mouse protein that serves as a transcriptional coactivator in yeast for the hormone binding domains of steroid receptors.
    Proc Natl Acad Sci U S A. 1996 May 14;93(10):4948-52 PMID: 8643509
  8. Molecular mechanisms of anti-inflammatory action of glucocorticoids.
    Bioessays. 1996 May;18(5):371-8 PMID: 8639160
  9. Suppression of interleukin-3-induced gene expression by a C-terminal truncated Stat5: role of Stat5 in proliferation.
    EMBO J. 1996 May 15;15(10):2425-33 PMID: 8665850
  10. IL-6-inducible complexes on an IL-6 response element of the junB promoter contain Stat3 and 36 kDa CRE-like site binding protein(s).
    Oncogene. 1996 Feb 1;12(3):547-54 PMID: 8637711
  11. Signaling by the cytokine receptor superfamily in normal and transformed hematopoietic cells.
    Adv Cancer Res. 1996;68:23-65 PMID: 8712070
  12. STAT3beta, a splice variant of transcription factor STAT3, is a dominant negative regulator of transcription.
    J Biol Chem. 1996 May 31;271(22):13221-7 PMID: 8675499
  13. Molecular determinants of glucocorticoid receptor function and tissue sensitivity to glucocorticoids.
    Endocr Rev. 1996 Jun;17(3):245-61 PMID: 8771358
  14. Mechanisms of transcriptional activation in vivo: two steps forward.
    Trends Genet. 1996 Aug;12(8):311-5 PMID: 8783941
  15. Characterization of mechanisms involved in transrepression of NF-kappa B by activated glucocorticoid receptors.
    Mol Cell Biol. 1995 Feb;15(2):943-53 PMID: 7823959
  16. Nuclear factor I and mammary gland factor (STAT5) play a critical role in regulating rat whey acidic protein gene expression in transgenic mice.
    Mol Cell Biol. 1995 Apr;15(4):2063-70 PMID: 7891701
  17. Requirement of serine phosphorylation for formation of STAT-promoter complexes.
    Science. 1995 Mar 31;267(5206):1990-4 PMID: 7701321
  18. Interleukin-3, granulocyte-macrophage colony stimulating factor and interleukin-5 transduce signals through two STAT5 homologs.
    EMBO J. 1995 Mar 15;14(6):1166-75 PMID: 7720707
  19. Interleukin-3 signals through multiple isoforms of Stat5.
    EMBO J. 1995 Apr 3;14(7):1402-11 PMID: 7537213
  20. Tyrosine-phosphorylated Stat1 and Stat2 plus a 48-kDa protein all contact DNA in forming interferon-stimulated-gene factor 3.
    Proc Natl Acad Sci U S A. 1995 Apr 25;92(9):3829-33 PMID: 7537377
  21. Prolactin, growth hormone, erythropoietin and granulocyte-macrophage colony stimulating factor induce MGF-Stat5 DNA binding activity.
    EMBO J. 1995 May 1;14(9):2005-13 PMID: 7744007
  22. Activation of Stat5 by interleukin 2 requires a carboxyl-terminal region of the interleukin 2 receptor beta chain but is not essential for the proliferative signal transmission.
    Proc Natl Acad Sci U S A. 1995 Jun 6;92(12):5482-6 PMID: 7777534
  23. Differentiation-regulated serine phosphorylation of STAT1 promotes GAF activation in macrophages.
    Mol Cell Biol. 1995 Jul;15(7):3579-86 PMID: 7791765
  24. Thrombopoietin activates a STAT5-like factor in hematopoietic cells.
    EMBO J. 1995 Jun 15;14(12):2847-56 PMID: 7796811
  25. Maximal activation of transcription by Stat1 and Stat3 requires both tyrosine and serine phosphorylation.
    Cell. 1995 Jul 28;82(2):241-50 PMID: 7543024
  26. CCAAT/enhancer-binding protein isoforms beta and delta are expressed in mammary epithelial cells and bind to multiple sites in the beta-casein gene promoter.
    J Biol Chem. 1995 Jul 28;270(30):17962-9 PMID: 7629103
  27. Signaling through the interleukin 2 receptor beta chain activates a STAT-5-like DNA-binding activity.
    Proc Natl Acad Sci U S A. 1995 Aug 1;92(16):7192-6 PMID: 7543676
  28. Negative cross-talk between RelA and the glucocorticoid receptor: a possible mechanism for the antiinflammatory action of glucocorticoids.
    Mol Endocrinol. 1995 Apr;9(4):401-12 PMID: 7659084
  29. Cloning and expression of Stat5 and an additional homologue (Stat5b) involved in prolactin signal transduction in mouse mammary tissue.
    Proc Natl Acad Sci U S A. 1995 Sep 12;92(19):8831-5 PMID: 7568026
  30. Cooperative transcriptional activity of Jun and Stat3 beta, a short form of Stat3.
    Proc Natl Acad Sci U S A. 1995 Sep 26;92(20):9097-101 PMID: 7568080
  31. Transcriptional responses to polypeptide ligands: the JAK-STAT pathway.
    Annu Rev Biochem. 1995;64:621-51 PMID: 7574495
  32. Developmentally and hormonally regulated CCAAT/enhancer-binding protein isoforms influence beta-casein gene expression.
    Mol Endocrinol. 1995 Sep;9(9):1223-32 PMID: 7491114
  33. The glucocorticoid receptor synergizes with Jun homodimers to activate AP-1-regulated promoters lacking GR binding sites.
    Chem Senses. 1995 Apr;20(2):251-5 PMID: 7583019
  34. Sequence and characterization of a coactivator for the steroid hormone receptor superfamily.
    Science. 1995 Nov 24;270(5240):1354-7 PMID: 7481822
  35. Stat1 depends on transcriptional synergy with Sp1.
    J Biol Chem. 1995 Dec 22;270(51):30264-7 PMID: 8530443
  36. Steroid hormone receptors: many actors in search of a plot.
    Cell. 1995 Dec 15;83(6):851-7 PMID: 8521509
  37. Prolactin-mediated gene activation in mammary epithelial cells.
    Curr Opin Genet Dev. 1995 Oct;5(5):587-94 PMID: 8664546
  38. Regulation of mammary gland factor/Stat5a during mammary gland development.
    Mol Endocrinol. 1995 Nov;9(11):1598-609 PMID: 8584036
  39. STATs: signal transducers and activators of transcription.
    Cell. 1996 Feb 9;84(3):331-4 PMID: 8608586
  40. Dominant negative stat3 mutant inhibits interleukin-6-induced Jak-STAT signal transduction.
    J Biol Chem. 1996 Mar 15;271(11):5961-4 PMID: 8626374
  41. Cooperation of Stat2 and p300/CBP in signalling induced by interferon-alpha.
    Nature. 1996 Sep 26;383(6598):344-7 PMID: 8848048
  42. Deletion of the carboxyl-terminal transactivation domain of MGF-Stat5 results in sustained DNA binding and a dominant negative phenotype.
    Mol Cell Biol. 1996 Oct;16(10):5691-700 PMID: 8816482
  43. Functional interactions between Stat5 and the glucocorticoid receptor.
    Nature. 1996 Oct 24;383(6602):726-8 PMID: 8878484
  44. Naturally occurring dominant negative variants of Stat5.
    Mol Cell Biol. 1996 Nov;16(11):6141-8 PMID: 8887644
  45. Nuclear receptor coactivators and corepressors.
    Mol Endocrinol. 1996 Oct;10(10):1167-77 PMID: 9121485
  46. An IL-2 response element in the human IL-2 receptor alpha chain promoter is a composite element that binds Stat5, Elf-1, HMG-I(Y) and a GATA family protein.
    EMBO J. 1996 Oct 15;15(20):5627-35 PMID: 8896456
  47. The general transcription factors of RNA polymerase II.
    Genes Dev. 1996 Nov 1;10(21):2657-83 PMID: 8946909
  48. Lactogenic hormone activation of Stat5 and transcription of the beta-casein gene in mammary epithelial cells is independent of p42 ERK2 mitogen-activated protein kinase activity.
    J Biol Chem. 1996 Dec 13;271(50):31863-8 PMID: 8943229
  49. Activation of Stat5a and Stat5b by tyrosine phosphorylation is tightly linked to mammary gland differentiation.
    Mol Endocrinol. 1996 Dec;10(12):1496-506 PMID: 8961260
  50. Two contact regions between Stat1 and CBP/p300 in interferon gamma signaling.
    Proc Natl Acad Sci U S A. 1996 Dec 24;93(26):15092-6 PMID: 8986769
  51. Stat5a is mandatory for adult mammary gland development and lactogenesis.
    Genes Dev. 1997 Jan 15;11(2):179-86 PMID: 9009201
  52. Nuclear integration of JAK/STAT and Ras/AP-1 signaling by CBP and p300.
    Proc Natl Acad Sci U S A. 1997 Feb 18;94(4):1074-9 PMID: 9037008
  53. Mechanism of gene expression by the glucocorticoid receptor: role of protein-protein interactions.
    Bioessays. 1997 Feb;19(2):153-60 PMID: 9046245
  54. Control by basal phosphorylation of cell cycle-dependent, hormone-induced glucocorticoid receptor hyperphosphorylation.
    Mol Endocrinol. 1997 Mar;11(3):305-11 PMID: 9058377
  55. Architectural transcription factors: proteins that remodel DNA.
    Cell. 1997 Mar 21;88(6):733-6 PMID: 9118214
  56. Transcriptional activation by recruitment.
    Nature. 1997 Apr 10;386(6625):569-77 PMID: 9121580
  57. The mammary gland: a unique organ for the study of development and tumorigenesis.
    J Mammary Gland Biol Neoplasia. 1996 Jan;1(1):5-19 PMID: 10887477
  58. Functional domains of the human glucocorticoid receptor.
    Cell. 1986 Aug 29;46(5):645-52 PMID: 3742595
  59. Functional domains of the human estrogen receptor.
    Cell. 1987 Dec 24;51(6):941-51 PMID: 3690665
  60. Prolactin regulation of beta-casein gene expression and of a cytosolic 120-kd protein in a cloned mouse mammary epithelial cell line.
    EMBO J. 1988 Jul;7(7):2089-95 PMID: 3416834
  61. Prolactin and glucocorticoid hormones synergistically induce expression of transfected rat beta-casein gene promoter constructs in a mammary epithelial cell line.
    Proc Natl Acad Sci U S A. 1989 Jan;86(1):104-8 PMID: 2643093
  62. Down-regulation and phosphorylation of glucocorticoid receptors in cultured cells. Investigations with a monospecific antiserum against a bacterially expressed receptor fragment.
    J Biol Chem. 1989 Aug 25;264(24):14396-402 PMID: 2668288
  63. Hormone-dependent phosphorylation of the glucocorticoid receptor occurs mainly in the amino-terminal transactivation domain.
    J Biol Chem. 1990 Apr 5;265(10):5403-8 PMID: 2108136
  64. Beta-casein gene promoter activity is regulated by the hormone-mediated relief of transcriptional repression and a mammary-gland-specific nuclear factor.
    Mol Cell Biol. 1991 Jul;11(7):3745-55 PMID: 2046676
  65. Epidermal growth factor receptor, platelet-derived growth factor receptor, and c-erbB-2 receptor activation all promote growth but have distinctive effects upon mouse mammary epithelial cell differentiation.
    Cell Growth Differ. 1991 Mar;2(3):145-54 PMID: 1676295
  66. Interference and synergism of glucocorticoid receptor and octamer factors.
    EMBO J. 1991 Sep;10(9):2513-21 PMID: 1714379
  67. The prolactin/growth hormone receptor family.
    Endocr Rev. 1991 Aug;12(3):235-51 PMID: 1935820
  68. Developmental and environmental regulation of a mammary gland-specific nuclear factor essential for transcription of the gene encoding beta-casein.
    Proc Natl Acad Sci U S A. 1992 Apr 1;89(7):3130-4 PMID: 1557422
  69. Interference between pathway-specific transcription factors: glucocorticoids antagonize phorbol ester-induced AP-1 activity without altering AP-1 site occupation in vivo.
    EMBO J. 1992 Jun;11(6):2241-6 PMID: 1318196
  70. Mutual cross-interference between glucocorticoid receptor and CREB inhibits transactivation in placental cells.
    New Biol. 1992 May;4(5):527-40 PMID: 1387550
  71. Kinetics of glucocorticoid receptor phosphorylation in intact cells. Evidence for hormone-induced hyperphosphorylation after activation and recycling of hyperphosphorylated receptors.
    J Biol Chem. 1993 Apr 15;268(11):7779-84 PMID: 8463306
  72. The basic region of AP-1 specifies glucocorticoid receptor activity at a composite response element.
    Genes Dev. 1992 Dec;6(12B):2491-501 PMID: 1340464
  73. Interaction of two sequence-specific single-stranded DNA-binding proteins with an essential region of the beta-casein gene promoter is regulated by lactogenic hormones.
    Mol Cell Biol. 1993 Dec;13(12):7303-10 PMID: 8246951
  74. The nuclear factor YY1 participates in repression of the beta-casein gene promoter in mammary epithelial cells and is counteracted by mammary gland factor during lactogenic hormone induction.
    Mol Cell Biol. 1994 Jan;14(1):128-37 PMID: 8264581
  75. Glucocorticoid receptor binding sites in the promoter region of milk protein genes.
    J Steroid Biochem Mol Biol. 1993 Dec;47(1-6):75-81 PMID: 8274444
  76. Physical association and functional antagonism between the p65 subunit of transcription factor NF-kappa B and the glucocorticoid receptor.
    Proc Natl Acad Sci U S A. 1994 Jan 18;91(2):752-6 PMID: 8290595
  77. Mammary gland factor (MGF) is a novel member of the cytokine regulated transcription factor gene family and confers the prolactin response.
    EMBO J. 1994 May 1;13(9):2182-91 PMID: 7514531
  78. Regulation of gene expression by prolactin.
    Rev Physiol Biochem Pharmacol. 1994;124:93-130 PMID: 8209140
Article Info
Journal
Molecular and cellular biology
Abbr.
Mol Cell Biol
ISSN
0270-7306
DOI
10.1128/MCB.17.11.6708
Published
1997-11-00
Pages
6708-16
Language
English
Region
United States
NLM ID
8109087
PMCID
PMC232525
Subset
IM
External Links
PubMed source DOI: 10.1128/MCB.17.11.6708 PMC Full Text View journal details
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