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PMID: 7784172 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

A modified oestrogen receptor ligand-binding domain as an improved switch for the regulation of heterologous proteins.

Nucleic acids research ·Vol. 23 ·No. 10 ·1995-05-25 ·Pages 1686-90

Littlewood TD, Hancock DC, Danielian PS, Parker MG, Evan GI

Abstract

A number of proteins have been rendered functionally oestrogen-dependent by fusion with the hormone-binding domain of the oestrogen receptor. There are, however, several significant disadvantages with such fusion proteins. First, their use in cells in vitro requires phenol red-free medium and laborious stripping of steroid hormones from serum in order to avoid constitutive activation. Secondly, control of oestrogen receptor fusion proteins in vivo is precluded by high endogenous levels of circulating oestrogens. Thirdly, the hormone-binding domain of the oestrogen receptor functions as a hormone-dependent transcriptional activation domain making interpretation of fusions with transcription factors problematical. In order to overcome these drawbacks we have used a transcriptionally inactive mutant of the murine oestrogen receptor which is unable to bind oestrogen yet retains normal affinity for the synthetic ligand, 4-hydroxytamoxifen. When the hormone-binding domain of this mutant oestrogen receptor is fused to the C-terminus of the c-Myc protein, Myc-induced proliferation and apoptosis in fibroblasts becomes dependent on 4-hydroxytamoxifen, but remains refractory to 17 beta-oestradiol.

MeSH Terms
3T3 Cells Amino Acid Sequence Animals Apoptosis/drug effects Base Sequence Binding Sites Cell Division/drug effects DNA Primers DNA-Binding Proteins/biosynthesis,metabolism Estradiol/pharmacology Gene Expression Gene Expression Regulation Kinetics Mice Molecular Sequence Data Mutagenesis, Site-Directed Point Mutation Polymerase Chain Reaction Proto-Oncogene Proteins c-myc/biosynthesis,metabolism Rats Receptors, Estrogen/biosynthesis,metabolism Recombinant Fusion Proteins/biosynthesis,metabolism Restriction Mapping Tamoxifen/pharmacology Transfection
Chemicals
DNA Primers DNA-Binding Proteins Proto-Oncogene Proteins c-myc Receptors, Estrogen Recombinant Fusion Proteins Tamoxifen Estradiol
Authors & Affiliations
5 authors, click to expand affiliations / ORCID
Littlewood T D
Molecular Endocrinology Laboratories, Imperial Cancer Research Fund, London, UK.
Hancock D C
Danielian P S
Parker M G
Evan G I
References (40)
40 references, click to expand
  1. Use of a conditional MyoD transcription factor in studies of MyoD trans-activation and muscle determination.
    Proc Natl Acad Sci U S A. 1993 Sep 1;90(17):8028-32 PMID: 8396258
  2. Effects of anti-estrogens on bone in castrated and intact female rats.
    Breast Cancer Res Treat. 1987 Oct;10(1):31-5 PMID: 3689979
  3. Modulation of cell proliferation and gene expression by a p53-estrogen receptor hybrid protein.
    Proc Natl Acad Sci U S A. 1993 Oct 15;90(20):9252-6 PMID: 8415687
  4. Interaction of hsp90 with steroid receptors: organizing some diverse observations and presenting the newest concepts.
    Mol Cell Endocrinol. 1990 Nov 12;74(1):C69-76 PMID: 2178103
  5. Chimaeras of myc oncoprotein and steroid receptors cause hormone-dependent transformation of cells.
    Nature. 1989 Jul 6;340(6228):66-8 PMID: 2662015
  6. 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):2811-8 PMID: 2118104
  7. Hormone-conditional transformation by fusion proteins of c-Abl and its transforming variants.
    EMBO J. 1993 Jul;12(7):2809-19 PMID: 8334997
  8. The MYC protein activates transcription of the alpha-prothymosin gene.
    EMBO J. 1991 Jan;10(1):133-41 PMID: 1989881
  9. CCAAT-enhancer binding protein: a component of a differentiation switch.
    Science. 1991 Jan 18;251(4991):288-92 PMID: 1987644
  10. The human estrogen receptor has two independent nonacidic transcriptional activation functions.
    Cell. 1989 Nov 3;59(3):477-87 PMID: 2805068
  11. Isolation of monoclonal antibodies specific for human c-myc proto-oncogene product.
    Mol Cell Biol. 1985 Dec;5(12):3610-6 PMID: 3915782
  12. Induction of apoptosis in fibroblasts by c-myc protein.
    Cell. 1992 Apr 3;69(1):119-28 PMID: 1555236
  13. Steroid-receptor fusion of the human immunodeficiency virus type 1 Rev transactivator: mapping cryptic functions of the arginine-rich motif.
    Proc Natl Acad Sci U S A. 1990 Oct;87(19):7787-91 PMID: 2217212
  14. Advanced mammalian gene transfer: high titre retroviral vectors with multiple drug selection markers and a complementary helper-free packaging cell line.
    Nucleic Acids Res. 1990 Jun 25;18(12):3587-96 PMID: 2194165
  15. c-Myc-induced apoptosis in fibroblasts is inhibited by specific cytokines.
    EMBO J. 1994 Jul 15;13(14):3286-95 PMID: 8045259
  16. Transcriptional activation and transformation by chimaeric Fos-estrogen receptor proteins: altered properties as a consequence of gene fusion.
    Oncogene. 1993 Oct;8(10):2781-90 PMID: 8378087
  17. Hormone-dependent transcriptional regulation and cellular transformation by Fos-steroid receptor fusion proteins.
    Proc Natl Acad Sci U S A. 1991 Jun 15;88(12):5114-8 PMID: 1711219
  18. Modulation of cyclin gene expression by adenovirus E1A in a cell line with E1A-dependent conditional proliferation.
    J Virol. 1994 Apr;68(4):2206-14 PMID: 8139005
  19. Identification of two transactivation domains in the mouse oestrogen receptor.
    Nucleic Acids Res. 1989 Jul 25;17(14):5477-88 PMID: 2762146
  20. Estrogen-dependent alterations in differentiation state of myeloid cells caused by a v-myb/estrogen receptor fusion protein.
    EMBO J. 1991 Dec;10(12):3713-9 PMID: 1718743
  21. Steroid receptors and their associated proteins.
    Mol Endocrinol. 1993 Jan;7(1):4-11 PMID: 8446107
  22. A selective transcriptional induction system for mammalian cells based on Gal4-estrogen receptor fusion proteins.
    Proc Natl Acad Sci U S A. 1993 Mar 1;90(5):1657-61 PMID: 8446579
  23. Hormone-regulated v-rel estrogen receptor fusion protein: reversible induction of cell transformation and cellular gene expression.
    EMBO J. 1992 Dec;11(12):4641-52 PMID: 1425595
  24. Endocrine pharmacology of antiestrogens as antitumor agents.
    Endocr Rev. 1990 Nov;11(4):578-610 PMID: 2292243
  25. The c-Myc protein induces cell cycle progression and apoptosis through dimerization with Max.
    EMBO J. 1993 Dec 15;12(13):5083-7 PMID: 8262051
  26. Conditional transformation of cells and rapid activation of the mitogen-activated protein kinase cascade by an estradiol-dependent human raf-1 protein kinase.
    Mol Cell Biol. 1993 Oct;13(10):6241-52 PMID: 8413224
  27. Localisation of the oestradiol-binding and putative DNA-binding domains of the human oestrogen receptor.
    EMBO J. 1986 Sep;5(9):2231-6 PMID: 3780678
  28. A movable and regulable inactivation function within the steroid binding domain of the glucocorticoid receptor.
    Cell. 1988 Sep 23;54(7):1073-80 PMID: 2843290
  29. Identification of residues in the estrogen receptor that confer differential sensitivity to estrogen and hydroxytamoxifen.
    Mol Endocrinol. 1993 Feb;7(2):232-40 PMID: 8469236
  30. Regulation of protein activities by fusion to steroid binding domains.
    Methods Cell Biol. 1994;43 Pt A:335-52 PMID: 7823870
  31. Steroid-binding domains for regulating the functions of heterologous proteins in cis.
    Trends Cell Biol. 1993 Aug;3(8):278-80 PMID: 14731747
  32. 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):2496-500 PMID: 3458212
  33. Contrasting actions of tamoxifen on endometrial and breast tumor growth in the athymic mouse.
    Cancer Res. 1988 Feb 15;48(4):812-5 PMID: 3338079
  34. Fusion of GAL4-VP16 to a steroid-binding domain provides a tool for gratuitous induction of galactose-responsive genes in yeast.
    Gene. 1993 Sep 6;131(1):129-34 PMID: 8370533
  35. Oncogenes and cell death.
    Curr Opin Genet Dev. 1994 Feb;4(1):120-9 PMID: 8193531
  36. The cloned human oestrogen receptor contains a mutation which alters its hormone binding properties.
    EMBO J. 1989 Jul;8(7):1981-6 PMID: 2792078
  37. The hormone-binding domains of the estrogen and glucocorticoid receptors contain an inducible transcription activation function.
    Cell. 1988 Jul 15;54(2):199-207 PMID: 3390864
  38. Biochemical pharmacology of antiestrogen action.
    Pharmacol Rev. 1984 Dec;36(4):245-76 PMID: 6395141
  39. Cooperative interaction between c-myc and bcl-2 proto-oncogenes.
    Nature. 1992 Oct 8;359(6395):554-6 PMID: 1406976
  40. Ectopic expression of a conditional GATA-2/estrogen receptor chimera arrests erythroid differentiation in a hormone-dependent manner.
    Genes Dev. 1993 Jun;7(6):1097-109 PMID: 8504932
Article Info
Journal
Nucleic acids research
Abbr.
Nucleic Acids Res
ISSN
0305-1048
DOI
10.1093/nar/23.10.1686
Published
1995-05-25
Pages
1686-90
Language
English
Region
England
NLM ID
0411011
PMCID
PMC306922
Subset
IM
External Links
PubMed source DOI: 10.1093/nar/23.10.1686 PMC Full Text View journal details
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