High affinity and specificity of dimeric binding of thyroid hormone receptors to DNA and their ligand-dependent dissociation.
The thyroid hormone receptors are ligand-dependent, DNA binding, trans-acting transcriptional factors belonging to the erbA-related steroid/thyroid hormone receptor superfamily. We report here the high affinity and specificity of dimeric DNA binding of human thyroid hormone receptor-alpha 1 (hTR alpha 1) and hTR beta 1 and the effect of T3 on its DNA binding. Gel mobility shift assay showed that hTR alpha 1 and -beta 1 bind to the rat GH-thyroid hormone response element (TRE) and rat malic enzyme (rME)-TRE as a monomer, dimer, and oligomer in the absence of T3 and bind to an irrelevant DNA sequence from the adenovirus 5 promoter only as a monomer. In competition studies using unlabeled TRE, dimer binding was displaced by lower concentrations of TRE than was the monomer, indicating that the dimeric binding has higher affinity than the monomer binding. These results suggest that the formation of dimers of TR increases the specificity and affinity for the response element. Surprisingly, T3 disrupted dimer binding and increased the intensity of monomer binding to rME-TRE in a dose-dependent manner. This does not occur with the rat GH-TRE. We also demonstrate that this disruption of dimeric binding by T3 occurred on a TRE formed by a direct repeat and not on a palindromic TRE. Furthermore, a mutant hTR beta (Mf), which has no detectable ligand-binding activity because of a glycine to arginine substitution at amino acid 340 in the hormone-binding domain, does not show dissociation from rME-TRE after the addition of T3.(ABSTRACT TRUNCATED AT 250 WORDS)
Miyamoto
T
T
Department of Medicine, University of Chicago, Illinois 60637.
Suzuki
S
S
DeGroot
L J
LJ
D000260:Adenoviruses, Human / Q000235:genetics; D000818:Animals; D001483:Base Sequence; D016384:Consensus Sequence; D004247:DNA / Q000378:metabolism*; D004279:DNA, Viral / Q000378:metabolism; D004268:DNA-Binding Proteins / Q000378:metabolism*; D013006:Growth Hormone / Q000235:genetics; D006801:Humans; D008024:Ligands; D008291:Malate Dehydrogenase / Q000235:genetics; D008969:Molecular Sequence Data; D011485:Protein Binding; D051381:Rats; D011988:Receptors, Thyroid Hormone / Q000378:metabolism*; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis; D014158:Transcription, Genetic; D014284:Triiodothyronine / Q000494:pharmacology
D004279:DNA, Viral; D004268:DNA-Binding Proteins; D008024:Ligands; D011988:Receptors, Thyroid Hormone; D011993:Recombinant Fusion Proteins; D014284:Triiodothyronine; D013006:Growth Hormone; D004247:DNA; D008291:Malate Dehydrogenase