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Title: Thyroid hormone receptors form distinct nuclear protein-dependent and independent complexes with a thyroid hormone response element

Abstract

We have examined the binding of nuclear proteins and recombinant thyroid hormone receptors (TRs) to the palindromic thyroid hormone responsive element AGGTCATGACCT (TREp) using a gel electrophoretic mobility shift assay. Four specific protein-DNA complexes were detected after incubation of nuclear extracts (NE) from T3-responsive pituitary (GH3) cells with a TREp-containing DNA fragment. This was compared with the TREp binding of reticulocyte lysate-synthesized TRs. TR alpha 1 and TR beta 2 each formed a single major TR:TREp complex which comigrated with the least retarded complex formed by GH3 NE, while TR beta 1 formed multiple complexes suggesting that it can bind to TREp as an oligomer. Interestingly, coincubation of 35S-TR alpha 1, GH3 NE, and unlabeled TREp resulted in not only the 35S-TR:TREp complex, but in two additional more greatly retarded complexes containing 35S-TR alpha 1 and comigrating with those formed by GH3 extract alone. Incubation of each of the TRs with NE from COS-7 cells, which do not possess sufficient endogenous TRs to mediate T3-responses, resulted in formation of a new, more greatly shifted complex. A similar, heat labile activity which altered mobility of the TR:TRE complex was also present in NE from T3-unresponsive JEG-3 cells. At high concentration ofmore » NE, all of the TR bound to TREp was more greatly retarded than in the absence of NE. Truncation of TR alpha 1 at amino acid 210 prevented additional complex formation in the presence of NE without affecting DNA binding, suggesting that the carboxyl-terminus of the TRs is essential for interaction with nuclear proteins.« less

Authors:
;  [1]
  1. Univ. of Pennsylvania School of Medicine, Philadelphia (USA)
Publication Date:
OSTI Identifier:
5895522
Resource Type:
Journal Article
Journal Name:
Molecular Endocrinology; (USA)
Additional Journal Information:
Journal Volume: 4:11; Journal ID: ISSN 0888-8809
Country of Publication:
United States
Language:
English
Subject:
59 BASIC BIOLOGICAL SCIENCES; NUCLEOPROTEINS; BIOCHEMICAL REACTION KINETICS; RECEPTORS; THYROID HORMONES; COMPLEXES; DNA; ELECTROPHORESIS; SULFUR 35; TRACER TECHNIQUES; BETA DECAY RADIOISOTOPES; BETA-MINUS DECAY RADIOISOTOPES; DAYS LIVING RADIOISOTOPES; EVEN-ODD NUCLEI; HORMONES; ISOTOPE APPLICATIONS; ISOTOPES; KINETICS; LIGHT NUCLEI; MEMBRANE PROTEINS; NUCLEI; NUCLEIC ACIDS; ORGANIC COMPOUNDS; PEPTIDE HORMONES; PROTEINS; RADIOISOTOPES; REACTION KINETICS; SULFUR ISOTOPES; 550201* - Biochemistry- Tracer Techniques

Citation Formats

Lazar, M A, and Berrodin, T J. Thyroid hormone receptors form distinct nuclear protein-dependent and independent complexes with a thyroid hormone response element. United States: N. p., 1990. Web. doi:10.1210/mend-4-11-1627.
Lazar, M A, & Berrodin, T J. Thyroid hormone receptors form distinct nuclear protein-dependent and independent complexes with a thyroid hormone response element. United States. https://doi.org/10.1210/mend-4-11-1627
Lazar, M A, and Berrodin, T J. 1990. "Thyroid hormone receptors form distinct nuclear protein-dependent and independent complexes with a thyroid hormone response element". United States. https://doi.org/10.1210/mend-4-11-1627.
@article{osti_5895522,
title = {Thyroid hormone receptors form distinct nuclear protein-dependent and independent complexes with a thyroid hormone response element},
author = {Lazar, M A and Berrodin, T J},
abstractNote = {We have examined the binding of nuclear proteins and recombinant thyroid hormone receptors (TRs) to the palindromic thyroid hormone responsive element AGGTCATGACCT (TREp) using a gel electrophoretic mobility shift assay. Four specific protein-DNA complexes were detected after incubation of nuclear extracts (NE) from T3-responsive pituitary (GH3) cells with a TREp-containing DNA fragment. This was compared with the TREp binding of reticulocyte lysate-synthesized TRs. TR alpha 1 and TR beta 2 each formed a single major TR:TREp complex which comigrated with the least retarded complex formed by GH3 NE, while TR beta 1 formed multiple complexes suggesting that it can bind to TREp as an oligomer. Interestingly, coincubation of 35S-TR alpha 1, GH3 NE, and unlabeled TREp resulted in not only the 35S-TR:TREp complex, but in two additional more greatly retarded complexes containing 35S-TR alpha 1 and comigrating with those formed by GH3 extract alone. Incubation of each of the TRs with NE from COS-7 cells, which do not possess sufficient endogenous TRs to mediate T3-responses, resulted in formation of a new, more greatly shifted complex. A similar, heat labile activity which altered mobility of the TR:TRE complex was also present in NE from T3-unresponsive JEG-3 cells. At high concentration of NE, all of the TR bound to TREp was more greatly retarded than in the absence of NE. Truncation of TR alpha 1 at amino acid 210 prevented additional complex formation in the presence of NE without affecting DNA binding, suggesting that the carboxyl-terminus of the TRs is essential for interaction with nuclear proteins.},
doi = {10.1210/mend-4-11-1627},
url = {https://www.osti.gov/biblio/5895522}, journal = {Molecular Endocrinology; (USA)},
issn = {0888-8809},
number = ,
volume = 4:11,
place = {United States},
year = {Thu Nov 01 00:00:00 EST 1990},
month = {Thu Nov 01 00:00:00 EST 1990}
}