Chemistry: molecular biology and microbiology – Micro-organism – tissue cell culture or enzyme using process... – Recombinant dna technique included in method of making a...
Patent
1993-05-10
1997-02-11
Ulm, John
Chemistry: molecular biology and microbiology
Micro-organism, tissue cell culture or enzyme using process...
Recombinant dna technique included in method of making a...
4352523, 4353201, 530350, 536 234, C12N 1562
Patent
active
056020090
DESCRIPTION:
BRIEF SUMMARY
FIELD OF THE INVENTION
The present invention relates to trans-repressing analog receptors of the steroid/thyroid superfamily. In a particular aspect, it relates to the identification and characterization of proteins that function as transcription trans-activation repressors, as well as to their preparation and use, including novel DNA isolates encoding same; expression vectors operatively harboring these DNA sequences; and hosts transfected with said vectors.
In another aspect, the present invention relates to the use of the above-described transcription trans-activation repressors in various assays and screening methods.
BACKGROUND OF THE INVENTION
The characterization and preparation of various hormone and hormone-like receptors, including steroid, thyroid, and retinoid receptors such as those represented by the glucocorticoid, mineralocorticoid, thyroid, estrogen-related and retinoid classes has been subject of considerable research.
It is known, for example, that the glucocorticoid receptor belongs to a large superfamily of ligand-dependent transcription factors that have themselves diverse roles in homeostasis, growth and development. Comparison of complementary DNAs encoding these receptors, as well as mutational analyses of their coding sequences, have identified certain functional domains within the molecule that are thought responsible respectively for DNA binding, hormone binding and nuclear localization. See Evans, et al., Science 240, 889 (1988) for a review of this subject matter.
In the case of the glucocorticoid receptor, the so-called DNA binding domain spans some sixty-six amino acids and is highly conserved among various species. In addition, this domain has been found to be required in order to activate transcription. See Hollenberg, et al., Cell 49, 39 (1987), Miesfeld, et al., Science 236, 423 (1987), Danielsen, et al., Mol.Endo 1, 816 (1987), Kumar, et al., Cell 51, 941 (1987), Gronemeyer, EMBO J. 6, 3985 (1987), and Waterman, et al., Mol.Endo 2, 14 (1988). This domain has been found to contain nine invariant cysteine residues. Although the contribution of each cysteine residue to overall function is unknown, as is the actual structure formed by this domain, it has been proposed that these cysteine residues coordinate two zinc ions to form two DNA binding, so-called finger domains, which result in a ternary structure thought responsible for the localization and binding of the glucocorticoid receptor to the requisite DNA site. See Klug, et al., Tr.Biochem.Sci 12, 464 (1987), Bens, et al., Cell 52, 1 (1988), and Evans, supra.
In a location nearer the carboxyl-terminal end distal from the DNA binding region is the so-called ligand binding domain which has the demonstrated ability to block activity of the receptor in the absence of hormone. Thus, presence of the requisite hormone relieves the inhibition of the receptor to activity. Deletion of this region has been found to produce a hormone-independent transcription activator. See Godowski, et al., Nature 325, 365 (1987), Hollenberg, et al., supra, Kumar, et al., supra, Danielsen et al., supra, and Adler et al., Cell, 52, 685 (1988).
In contrast to these two domains, the sequences lying towards the amino-terminal region from the DNA binding domain are poorly understood both as to structure, and particularly, function. This region is extremely variable both in size and in composition amongst the various receptors--See Evans, supra--and may contribute to the heterogeneity of receptor function. See Kumar et al., supra, and Tora et al., 333, 185 (1988).
Despite extensive analysis, some of which has been reported in the scientific literature, the region(s) that determine(s) trans-activation of transcription initiation remains poorly characterized. Trans-activation domains can be defined as polypeptide regions that, when combined with the functional DNA binding domain, increase productive transcription initiation by RNA polymerases. See Sigler, Nature 333, 210 (1988), Brent et al., Cell 43, 729 (1985), Hope et al., Cell 46, 885 (1986), Ma e
REFERENCES:
patent: 4818684 (1989-04-01), Edelman et al.
Adler et al., "Steroid Receptor-Mediated Inhibition of Rat Prolactin Gene Expression Does Not Require the Receptor DNA-Binding Domain" Cell 52:685-695 (1988).
Nature 330:624-629, 17 Dec. 1987, Gigisene et al Identification of a receptor for the morphogen retilore acids.
Akerblom et al., "Negative Regulation by Glucocorticoids Through Interference with cAMP Responsive Enhancer" Science 241:350-353 (1988).
Anderson and Axel, "A Bipotential Neuroendocrine Precursor Whose Choice of Cell Fate Is Determined by NGF and Glucocorticoids" Cell 47:1079-1090 (1986).
Berg, Jeremy M., "DNA Binding Specificity of Steroid Receptors" Cell 57:1065-1068 (1989).
Brent et al., "Thyroid Hormone Aporeceptor Represses T3-Inducible Promoters and Blocks Activity of the Retinoic Acid Receptor" The New Biologist 1(3):329-336 (1989).
Brent and Ptashne, "A Eukaryotic Transcriptional Activator Bearing the DNA Specificity of a Prokaryotic Repressor" Cell 43:729-736 (1985).
Clark et al., "Pharmaceuticals from transgenic livestock" Tibtech 5:20-24 (1987).
Damm et al., "Protein encoded by v-erbA functions as a thyroid-hormone receptor antagonist" Nature 339(6226):593-597 (1989).
Damm et al., "A single point mutation in erbA restores the erythroid transforming potential of a mutant avian erythroblastosis virus (AEV) defective in both erbA and erbB oncogenes" EMBO J. 6(2):375-382 (1987).
Danielsen et al., "Domains of the Glucocorticoid Receptor Involved in Specific and Nonspecific Deoxyribonucleic Acid Binding, Hormone Activation, and Transcriptional Enhancement" Mol. Endo. 1(11):816-822 (1987).
Delegeane et al., "Tissue-Specific Enhancer of the Human Glycoprotein Hormone .alpha.-Subunit Gene: Dependence on Cyclic AMP-Inducible Elements" Molecular and Cellular Biol. 7(11):3994-4002 (1987).
deWet et al., "Firefly Luciferase Gene: Structure and Expression in Mammalian Cells" Molecular and Cellular Biol. 7(2):725-737 (1987).
Espeseth et al., "Retinoic acid receptor expression vector inhibits differentiation of F9 embryonal carcinoma cells" Genes & Development 3:1647-1656 (1989).
Evans, Ronald M., "The Steroid and Thyroid Hormone Receptor Superfamily" Science 240:889-895 (1988).
Evans and Hollenberg, "Zinc Fingers: Gilt by Association" Cell 52:1-3 (1988).
Giguere et al., "Functional Domains of the Human Glucocorticoid Receptor" Cell 46:645-652 (1986).
Glass et al., "The Thyroid Hormone Receptor Binds with Opposite Transcriptional Effects to a Common Sequence Motif in Thyroid Hormone and Estrogen Response Elements" Cell 54:313-323 (1988).
Glass et al., "A c-erb-A binding site in rat growth hormone gene mediates trans-activation by thyroid hormone" Nature 329:738-741 (1987).
Godowski et al., "Signal Transduction and Transcriptional Regulation by Glucocorticoid Receptor-LexA Fusion Proteins" Science 241:812-816 (1988).
Graupner et al., "Dual regulatory role for thyroid-hormone receptors allows control of retinoic-acid receptor activity" Nature 340:653-656 (1989).
Green and Chambon, "Oestradiol induction of a glucocorticoid-responsive gene by a chimaeric receptor" Nature 325:74-78 (1987).
Gronemeyer et al., "The chicken progesterone receptor: sequence, expression and functional analysis" EMBO J. 6:3985-3994 (1987).
Herskowitz, Ira, "Functional inactivation of genes by dominant negative mutations" Nature 329:219-222 (1987).
Hollenberg and Evans, "Multiple and Cooperative Trans-Activation Domains of the Human Glucocorticoid Receptor" Cells 55:899-906 (1988).
Hollenberg et al., "Colocalization of DNA-Binding and Transcriptional Activation Functions in the Human Glucocorticoid Receptor" Cell 49:39-46 (1987).
Hope et al., "Structural and functional characterizations of the short acidic transcriptional activation region of yeast GCN4 protein" Nature 333:635-640 (1988).
Hope and Struhl, "Functional Dissection of a Eukaryotic Transcriptional Activator Protein, GCN4 of Yeast" Yeast 46:885-894 (1986).
Hu and Gudas, "Cyclic AMP Analogs and Retinoic Acid I
Damm Klaus
Evans Ronald M.
Heyman Richard A.
Hollenberg Stanley M.
Oro Anthony E.
Ramos Robert T.
Reiter Stephen E.
The Salk Institute for Biological Studies
Ulm John
LandOfFree
Dominant negative chimeras of the steroid/thyroid superfamily of does not yet have a rating. At this time, there are no reviews or comments for this patent.
If you have personal experience with Dominant negative chimeras of the steroid/thyroid superfamily of, we encourage you to share that experience with our LandOfFree.com community. Your opinion is very important and Dominant negative chimeras of the steroid/thyroid superfamily of will most certainly appreciate the feedback.
Profile ID: LFUS-PAI-O-341218