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Mol Med. 1995 March; 1(3): 306–319.
PMCID: PMC2229906

Cell type-dependent modulation of the dominant negative action of human mutant thyroid hormone beta 1 receptors.


BACKGROUND: Mutations in the ligand-binding domain of the thyroid hormone receptor beta (TR beta) gene cause the syndrome of resistance to thyroid hormone (RTH). The clinical phenotype results from the antagonism of the normal TR alpha and the non-mutated TR beta alleles by the TR beta 1 mutants, via a dominant negative effect. There is, however, marked heterogeneity of organ resistance within and among kindreds with RTH. This study examines the potential role of cell type in modulating the dominant negative potency of human TR beta 1 (h-TR beta 1) mutants. MATERIALS AND METHODS: Transient transfections were performed in HeLa and NIH3T3 cells, using a wild type (WT) and three naturally occurring mutant h-TR beta 1 constructs, and three natural thyroid hormone response elements (TREs). Immunocytochemistry was performed to detect levels of TR beta 1 expression in these two cell types. In order to determine how TR beta 1 interacts with other cellular partners, gel-shift analyses using HeLa and NIH3T3 nuclear extracts were performed. RESULTS: Transfection studies using WT h-TR beta 1 in HeLa and NIH3T3 cells, showed that the 3,3',5-triiodothyronine (T3)-induced transactivation of the different TREs varied between cell types. Unlike the non-T3-binding h-TR beta 1 mutant, PV, mutants ED and OK displayed the expected T3-induced dose responsiveness in these two cell types. For each TRE examined, the magnitude of the dominant negative effect varied between the cell types. The levels of receptor expression in HeLa and NIH3T3 cells were identical, as determined by immunocytochemistry. Gel-shift analyses showed differences in the formation of hetero- and homodimers depending on both the cell type and TRE motif. CONCLUSIONS: The cell type in which a mutant receptor operates affects the relative amounts of hetero- and homodimers. Together with the nature of the mutation and the TRE-motif, this could modulate the dominant negative action of mutant receptors in different tissues, which, in turn, could contribute to the variable phenotypic characteristics of RTH.

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Selected References

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  • Refetoff S, DeWind LT, DeGroot LJ. Familial syndrome combining deaf-mutism, stuppled epiphyses, goiter and abnormally high PBI: possible target organ refractoriness to thyroid hormone. J Clin Endocrinol Metab. 1967 Feb;27(2):279–294. [PubMed]
  • Refetoff S, Weiss RE, Usala SJ. The syndromes of resistance to thyroid hormone. Endocr Rev. 1993 Jun;14(3):348–399. [PubMed]
  • Hauser P, Zametkin AJ, Martinez P, Vitiello B, Matochik JA, Mixson AJ, Weintraub BD. Attention deficit-hyperactivity disorder in people with generalized resistance to thyroid hormone. N Engl J Med. 1993 Apr 8;328(14):997–1001. [PubMed]
  • Usala SJ, Bale AE, Gesundheit N, Weinberger C, Lash RW, Wondisford FE, McBride OW, Weintraub BD. Tight linkage between the syndrome of generalized thyroid hormone resistance and the human c-erbA beta gene. Mol Endocrinol. 1988 Dec;2(12):1217–1220. [PubMed]
  • Parrilla R, Mixson AJ, McPherson JA, McClaskey JH, Weintraub BD. Characterization of seven novel mutations of the c-erbA beta gene in unrelated kindreds with generalized thyroid hormone resistance. Evidence for two "hot spot" regions of the ligand binding domain. J Clin Invest. 1991 Dec;88(6):2123–2130. [PMC free article] [PubMed]
  • Nagaya T, Madison LD, Jameson JL. Thyroid hormone receptor mutants that cause resistance to thyroid hormone. Evidence for receptor competition for DNA sequences in target genes. J Biol Chem. 1992 Jun 25;267(18):13014–13019. [PubMed]
  • Nagaya T, Jameson JL. Thyroid hormone receptor dimerization is required for dominant negative inhibition by mutations that cause thyroid hormone resistance. J Biol Chem. 1993 Jul 25;268(21):15766–15771. [PubMed]
  • Chatterjee VK, Nagaya T, Madison LD, Datta S, Rentoumis A, Jameson JL. Thyroid hormone resistance syndrome. Inhibition of normal receptor function by mutant thyroid hormone receptors. J Clin Invest. 1991 Jun;87(6):1977–1984. [PMC free article] [PubMed]
  • Yen PM, Sugawara A, Refetoff S, Chin WW. New insights on the mechanism(s) of the dominant negative effect of mutant thyroid hormone receptor in generalized resistance to thyroid hormone. J Clin Invest. 1992 Nov;90(5):1825–1831. [PMC free article] [PubMed]
  • Weiss RE, Weinberg M, Refetoff S. Identical mutations in unrelated families with generalized resistance to thyroid hormone occur in cytosine-guanine-rich areas of the thyroid hormone receptor beta gene. Analysis of 15 families. J Clin Invest. 1993 Jun;91(6):2408–2415. [PMC free article] [PubMed]
  • Burnside J, Darling DS, Chin WW. A nuclear factor that enhances binding of thyroid hormone receptors to thyroid hormone response elements. J Biol Chem. 1990 Feb 15;265(5):2500–2504. [PubMed]
  • Murray MB, Towle HC. Identification of nuclear factors that enhance binding of the thyroid hormone receptor to a thyroid hormone response element. Mol Endocrinol. 1989 Sep;3(9):1434–1442. [PubMed]
  • Kliewer SA, Umesono K, Mangelsdorf DJ, Evans RM. Retinoid X receptor interacts with nuclear receptors in retinoic acid, thyroid hormone and vitamin D3 signalling. Nature. 1992 Jan 30;355(6359):446–449. [PubMed]
  • Yu VC, Delsert C, Andersen B, Holloway JM, Devary OV, När AM, Kim SY, Boutin JM, Glass CK, Rosenfeld MG. RXR beta: a coregulator that enhances binding of retinoic acid, thyroid hormone, and vitamin D receptors to their cognate response elements. Cell. 1991 Dec 20;67(6):1251–1266. [PubMed]
  • Zhang XK, Hoffmann B, Tran PB, Graupner G, Pfahl M. Retinoid X receptor is an auxiliary protein for thyroid hormone and retinoic acid receptors. Nature. 1992 Jan 30;355(6359):441–446. [PubMed]
  • Leid M, Kastner P, Lyons R, Nakshatri H, Saunders M, Zacharewski T, Chen JY, Staub A, Garnier JM, Mader S, et al. Purification, cloning, and RXR identity of the HeLa cell factor with which RAR or TR heterodimerizes to bind target sequences efficiently. Cell. 1992 Jan 24;68(2):377–395. [PubMed]
  • Mixson AJ, Parrilla R, Ransom SC, Wiggs EA, McClaskey JH, Hauser P, Weintraub BD. Correlations of language abnormalities with localization of mutations in the beta-thyroid hormone receptor in 13 kindreds with generalized resistance to thyroid hormone: identification of four new mutations. J Clin Endocrinol Metab. 1992 Oct;75(4):1039–1045. [PubMed]
  • Mixson AJ, Hauser P, Tennyson G, Renault JC, Bodenner DL, Weintraub BD. Differential expression of mutant and normal beta T3 receptor alleles in kindreds with generalized resistance to thyroid hormone. J Clin Invest. 1993 May;91(5):2296–2300. [PMC free article] [PubMed]
  • Meier CA, Dickstein BM, Ashizawa K, McClaskey JH, Muchmore P, Ransom SC, Menke JB, Hao EH, Usala SJ, Bercu BB, et al. Variable transcriptional activity and ligand binding of mutant beta 1 3,5,3'-triiodothyronine receptors from four families with generalized resistance to thyroid hormone. Mol Endocrinol. 1992 Feb;6(2):248–258. [PubMed]
  • Williams GR, Harney JW, Forman BM, Samuels HH, Brent GA. Oligomeric binding of T3 receptor is required for maximal T3 response. J Biol Chem. 1991 Oct 15;266(29):19636–19644. [PubMed]
  • Hao E, Menke JB, Smith AM, Jones C, Geffner ME, Hershman JM, Wuerth JP, Samuels HH, Ways DK, Usala SJ. Divergent dimerization properties of mutant beta 1 thyroid hormone receptors are associated with different dominant negative activities. Mol Endocrinol. 1994 Jul;8(7):841–851. [PubMed]
  • Au-Fliegner M, Helmer E, Casanova J, Raaka BM, Samuels HH. The conserved ninth C-terminal heptad in thyroid hormone and retinoic acid receptors mediates diverse responses by affecting heterodimer but not homodimer formation. Mol Cell Biol. 1993 Sep;13(9):5725–5737. [PMC free article] [PubMed]
  • Martin KJ. The interactions of transcription factors and their adaptors, coactivators and accessory proteins. Bioessays. 1991 Oct;13(10):499–503. [PubMed]
  • Williams GR, Harney JW, Moore DD, Larsen PR, Brent GA. Differential capacity of wild type promoter elements for binding and trans-activation by retinoic acid and thyroid hormone receptors. Mol Endocrinol. 1992 Oct;6(10):1527–1537. [PubMed]
  • Flug F, Copp RP, Casanova J, Horowitz ZD, Janocko L, Plotnick M, Samuels HH. cis-acting elements of the rat growth hormone gene which mediate basal and regulated expression by thyroid hormone. J Biol Chem. 1987 May 5;262(13):6373–6382. [PubMed]
  • Steinfelder HJ, Hauser P, Nakayama Y, Radovick S, McClaskey JH, Taylor T, Weintraub BD, Wondisford FE. Thyrotropin-releasing hormone regulation of human TSHB expression: role of a pituitary-specific transcription factor (Pit-1/GHF-1) and potential interaction with a thyroid hormone-inhibitory element. Proc Natl Acad Sci U S A. 1991 Apr 15;88(8):3130–3134. [PubMed]
  • Schreiber E, Matthias P, Müller MM, Schaffner W. Rapid detection of octamer binding proteins with 'mini-extracts', prepared from a small number of cells. Nucleic Acids Res. 1989 Aug 11;17(15):6419–6419. [PMC free article] [PubMed]
  • Lin KH, Willingham MC, Liang CM, Cheng SY. Intracellular distribution of the endogenous and transfected beta form of thyroid hormone nuclear receptor visualized by the use of domain-specific monoclonal antibodies. Endocrinology. 1991 May;128(5):2601–2609. [PubMed]
  • Sugawara A, Yen PM, Darling DS, Chin WW. Characterization and tissue expression of multiple triiodothyronine receptor-auxiliary proteins and their relationship to the retinoid X-receptors. Endocrinology. 1993 Sep;133(3):965–971. [PubMed]
  • Zavacki AM, Harney JW, Brent GA, Larsen PR. Dominant negative inhibition by mutant thyroid hormone receptors is thyroid hormone response element and receptor isoform specific. Mol Endocrinol. 1993 Oct;7(10):1319–1330. [PubMed]
  • Suen CS, Yen PM, Chin WW. In vitro transcriptional studies of the roles of the thyroid hormone (T3) response elements and minimal promoters in T3-stimulated gene transcription. J Biol Chem. 1994 Jan 14;269(2):1314–1322. [PubMed]
  • Forman BM, Samuels HH. Interactions among a subfamily of nuclear hormone receptors: the regulatory zipper model. Mol Endocrinol. 1990 Sep;4(9):1293–1301. [PubMed]
  • Forman BM, Yang CR, Au M, Casanova J, Ghysdael J, Samuels HH. A domain containing leucine-zipper-like motifs mediate novel in vivo interactions between the thyroid hormone and retinoic acid receptors. Mol Endocrinol. 1989 Oct;3(10):1610–1626. [PubMed]
  • Darling DS, Beebe JS, Burnside J, Winslow ER, Chin WW. 3,5,3'-triiodothyronine (T3) receptor-auxiliary protein (TRAP) binds DNA and forms heterodimers with the T3 receptor. Mol Endocrinol. 1991 Jan;5(1):73–84. [PubMed]
  • O'Donnell AL, Koenig RJ. Mutational analysis identifies a new functional domain of the thyroid hormone receptor. Mol Endocrinol. 1990 May;4(5):715–720. [PubMed]
  • O'Donnell AL, Rosen ED, Darling DS, Koenig RJ. Thyroid hormone receptor mutations that interfere with transcriptional activation also interfere with receptor interaction with a nuclear protein. Mol Endocrinol. 1991 Jan;5(1):94–99. [PubMed]
  • Spanjaard RA, Darling DS, Chin WW. Ligand-binding and heterodimerization activities of a conserved region in the ligand-binding domain of the thyroid hormone receptor. Proc Natl Acad Sci U S A. 1991 Oct 1;88(19):8587–8591. [PubMed]
  • Geffner ME, Su F, Ross NS, Hershman JM, Van Dop C, Menke JB, Hao E, Stanzak RK, Eaton T, Samuels HH, et al. An arginine to histidine mutation in codon 311 of the C-erbA beta gene results in a mutant thyroid hormone receptor that does not mediate a dominant negative phenotype. J Clin Invest. 1993 Feb;91(2):538–546. [PMC free article] [PubMed]
  • Dulgeroff AJ, Geffner ME, Koyal SN, Wong M, Hershman JM. Bromocriptine and Triac therapy for hyperthyroidism due to pituitary resistance to thyroid hormone. J Clin Endocrinol Metab. 1992 Oct;75(4):1071–1075. [PubMed]

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