Divergent dimerization properties of mutant beta 1 thyroid hormone receptors are associated with different dominant negative activities
| Autor: | J B Menke, C Jones, Stephen J. Usala, H H Samuels, Mitchell E. Geffner, A M Smith, E Hao, J P Wuerth, D K Ways, J M Hershman |
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| Rok vydání: | 1994 |
| Předmět: |
Thyroid Hormone Receptor Gene
medicine.medical_specialty Protein Conformation Molecular Sequence Data Biology Protein Structure Secondary Cell Line Thyroid hormone receptor beta Endocrinology Internal medicine Chlorocebus aethiops medicine Animals Humans Point Mutation Amino Acid Sequence Molecular Biology Genes Dominant Receptors Thyroid Hormone Thyroid hormone receptor Base Sequence Thyroid DNA General Medicine medicine.anatomical_structure Nuclear receptor Thyroid hormone receptor alpha Hormone receptor Triiodothyronine Hormone |
| Zdroj: | Molecular Endocrinology. 8:841-851 |
| ISSN: | 1944-9917 0888-8809 |
| DOI: | 10.1210/mend.8.7.7984146 |
| Popis: | Syndromes of resistance to thyroid hormones are caused by mutations in the T3-binding domain of the c-erbA beta thyroid hormone receptor gene. The S receptor (deletion of THR332) is a potent dominant negative protein cloned from a kindred with generalized resistance to thyroid hormones. The G-H receptor (ARG311HIS) has compromised dominant negative function and was found in both normal individuals and in a patient with severe pituitary resistance to thyroid hormones. We have investigated the mechanism responsible for the difference in receptor phenotypes by analyzing the binding of S and G-H receptors to thyroid hormone response elements with electrophoretic mobility shift analysis. Wild-type human c-erbA beta 1 (WT), S, and G-H receptors were synthesized in reticulocyte lysate, reacted with a thyroid hormone response element consisting of a direct repeat with 4 base pairs (DR+4; AGGTCA CAGG AGGTCA), and the products analyzed by gel shift. G-H receptor homodimerization was greatly impaired; G-H formed predominantly monomeric complex compared with monomeric and homodimeric WT complexes. The G-H receptor was able to form heterodimeric complexes with cellular thyroid hormone receptor auxiliary protein (TRAP) factors including the human retinoid X receptor-alpha. When TRAP was limiting, the levels of G-H heterodimeric complex were 2- to 3-fold reduced compared with WT receptor. In contrast to the WT and G-H receptors, the S receptor formed almost exclusively homodimeric complex with DR+4; the approximate ratio of S:WT:G-H homodimeric complexes at equivalent concentrations of receptors was 60:20:1. A measurable increase (1.2- to 2.6-fold) in heterodimeric complex formation was observed with the S receptor relative to WT when TRAP was at limiting concentration. As reported previously by others, thyroid hormone significantly reduced the WT homodimeric complex with DR+4. There was no effect on the S homodimeric complex. Finally, the WT, S, and G-H receptors formed different complexes with the element consisting of an inverted repeat with 5 base pairs (IR+5; AGGTCA ACAGT TGACCT) and the IR element (AGGTCA TGACCT), which were differently regulated by thyroid hormone. The S receptor bound as a homodimer with IR+5, whereas the WT receptor bound as a homodimer only with thyroid hormone. No homodimeric complex formed with IR+5 and the G-H receptor. Qualitatively similar results were observed with the IR element. We conclude that the ARG311HIS mutation severely perturbs the homodimerization and, to a much less degree, heterodimerization functions of the c-erbA beta 1 receptor. Furthermore, the THR332 deletion mutation augments homodimerization of the c-erbA beta 1 receptor. These results indicate that different mutations in the c-erbA beta 1 thyroid hormone receptor have divergently affected dimerization activities which seem to influence the level of dominant negative activity in man. |
| Databáze: | OpenAIRE |
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