Fatty acyl-CoA binding activity of the nuclear thyroid hormone receptor

1993 ◽  
Vol 51 (4) ◽  
pp. 458-464 ◽  
Author(s):  
Qiulin Li ◽  
Naoki Yamamoto ◽  
Seiji Morisawa ◽  
Akira Inoue
Keyword(s):  
Thyroid Hormone ◽  
Hormone Receptor ◽  
Thyroid Hormone Receptor ◽  
Fatty Acyl ◽  
Binding Activity

Fatty Acyl-CoAs Are Potent Inhibitors of the Nuclear Thyroid Hormone Receptor In Vitro

1990 ◽  
Vol 107 (5) ◽  
pp. 699-702 ◽  
Author(s):  
Qiulin Li ◽  
Naoki Yamamoto ◽  
Akira Inoue ◽  
Seiji Morisawa
Keyword(s):  
Thyroid Hormone ◽  
Hormone Receptor ◽  
Thyroid Hormone Receptor ◽  
Fatty Acyl

Purification of human c-erb A β protein

1991 ◽  
Vol 7 (2) ◽  
pp. 123-129 ◽  
Author(s):  
K. Ichikawa ◽  
K. Hashizume ◽  
Y. Nishii ◽  
T. Takeda ◽  
M. Kobayashi ◽  
...  
Keyword(s):  
Thyroid Hormone ◽  
Column Chromatography ◽  
Hormone Receptor ◽  
Thyroid Hormone Receptor ◽  
Gel Filtration ◽  
Binding Activity ◽  
Human Thyroid ◽  
E Coli ◽  
Ammonium Sulphate Precipitation ◽  
Β Protein

ABSTRACT Human thyroid hormone receptor (c-erb A protein) produced by Escherichia coli expression vector plasmid was purified sequentially using polyethylenimine precipitation of DNA, hydroxylapatite column chromatography, ammonium sulphate precipitation, Sephacryl S-300 gel filtration and mono Q-Sepharose column chromatography. These column procedures resulted in 41.3-fold purification of 3,5,3′-tri-iodo-l-thyronine (T3) binding activity over the initial E. coli extract. Purified protein as well as crude preparation showed high-affinity binding to T3. The c-erb A protein enriched by column purification was further purified by electroelution after electrophoresis. Rabbit antibody against the c-erb A protein was prepared and used for the Western blotting analysis. The antibody recognized c-erb A protein but not the bacterial proteins in crude E. coli extract. When partially purified rat hepatic nuclear thyroid hormone receptor was analysed, a 56kDa receptor was specifically recognized by the antibody.

scholarly journals Loss of thyroid hormone receptor activity in primary cultured rat hepatocytes is reversed by 2-mercaptoethanol

1992 ◽  
Vol 281 (3) ◽  
pp. 669-673 ◽  
Author(s):  
N Yamamoto ◽  
A Inoue ◽  
K P Takahashi ◽  
Q Li ◽  
H Nakamura ◽  
...  
Keyword(s):  
Thyroid Hormone ◽  
Hormone Receptor ◽  
Hormone Receptors ◽  
Thyroid Hormone Receptor ◽  
Primary Cultures ◽  
Rat Hepatocytes ◽  
Binding Activity ◽  
Dependent Manner ◽  
Hormone Binding ◽  
Level Of Activity

In primary cultures of rat hepatocytes, specific thyroid-hormone-binding activity diminished with time and was hardly detectable at 24 h. In accordance with the loss of 3,5,3′-tri-iodothyronine (T3) binding, responses to the hormone disappeared, as indicated by low induction of the thyroid-hormone-responsive gene S14. In contrast, thyroid hormone receptor proteins were present, as determined by immunostaining with a specific antibody against the receptor. Thus the loss of T3 binding was due to receptor inactivation. After various attempts to restore the T3-binding activity, we found that 2-mercaptoethanol, a reducing agent, when added to the culture medium restored the hormone binding activity in a dose- and time-dependent manner. The observed kinetics and experiments using cycloheximide suggested that mercaptoethanol prevented inactivation of the newly synthesized receptors. Oxidoreductive conditions within cells may have a role in determining the level of activity of thyroid hormone receptors.

Positive and negative modulation of Jun action by thyroid hormone receptor at a unique AP1 site

1993 ◽  
Vol 13 (5) ◽  
pp. 3042-3049
Author(s):  
G Lopez ◽  
F Schaufele ◽  
P Webb ◽  
J M Holloway ◽  
J D Baxter ◽  
...  
Keyword(s):  
Thyroid Hormone ◽  
Transcriptional Activation ◽  
Hormone Receptor ◽  
Thyroid Hormone Receptor ◽  
Binding Activity ◽  
Binding Domain ◽  
Dna Binding Domain ◽  
Plasmid Sequence ◽  
Regulatory Effects

We have characterized the putative AP1 site in the backbone of pUC plasmids and found unique regulatory effects. The site, which mapped to a 19-bp region around nucleotide 37, conferred transcriptional activation by Jun or Jun/Fos that was boosted up to fivefold by unliganded thyroid hormone receptor (TR). Thyroid hormone changed potentiation of the Jun response by TR into repression. Although the plasmid sequence is a near-perfect consensus AP1 site, the perfect consensus AP1 site from the human collagenase promoter did not show the same effects. Deletion of the ligand binding domain of the TR eliminated the ability of the receptor to boost Jun activity, and deletion, mutation, or changes in specificity of the DNA binding domain eliminated both its ability to potentiate Jun activity and repress with hormone. In vitro Jun/Fos complexes bound the operative plasmid fragment, and the presence of TR interfered very little with Jun/Fos binding activity. Protein interaction studies in the absence of DNA showed that TR bound Jun protein in solution either in the presence or in the absence of hormone. These observations suggest a mechanism for synergy and repression by TR through modulation of Jun activity: positive when TR is unliganded, and negative when hormone is bound. They also suggest that the presence of the plasmid element can confound studies of the regulation of linked promoters.

Secretory expression of thyroid hormone receptor using transgenic silkworms and its DNA binding activity

2020 ◽  
Vol 176 ◽  
pp. 105723
Author(s):  
Hirofumi Nakaya ◽  
Ken-ichiro Tatematsu ◽  
Hideki Sezutsu ◽  
Nobuo Kuwabara ◽  
Noriyuki Koibuchi ◽  
...  
Keyword(s):  
Thyroid Hormone ◽  
Dna Binding ◽  
Hormone Receptor ◽  
Thyroid Hormone Receptor ◽  
Binding Activity ◽  
Secretory Expression ◽  
Dna Binding Activity ◽  
Transgenic Silkworms

Interaction of sodium molybdate with the thyroid hormone receptor

1990 ◽  
Vol 68 (3) ◽  
pp. 630-634 ◽  
Author(s):  
Robert Faure ◽  
Jean H. Dussault
Keyword(s):  
Thyroid Hormone ◽  
Hormone Receptor ◽  
Thyroid Hormone Receptor ◽  
Sodium Molybdate ◽  
Binding Activity ◽  
Exchange Chromatography ◽  
Receptor Interaction ◽  
Affinity Constant ◽  
Receptor Complexes ◽  
Receptor Binding Activity

The 3,5,3′-triiodothyronine (T3) binding activity of solubilized nuclear proteins from rat liver was decreased when molybdate (10 mM) was present in the incubation medium in the absence of thiol reagents. The equilibrium affinity constant was reduced by 40%. The rate of degradation of T3-receptor complexes at 37 °C remained unchanged, but when the extracts were further reincubated in the presence of β-mercaptoethanol, molybdate had a protective effect after 5 h incubation at 37 °C. In contrast, the thyroxine (T4) binding activity was not affected by heating at 37 °C or by molybdate. Ion-exchange chromatography confirmed the existence of a molybdate–receptor interaction: the T3–receptor complexes shifted from elution at 0.22 to 0.20 M NaCl with the progressive appearance of a small leader peak, whereas the T4-receptor complexes eluted in a large and split peak (0.22–0.4 M NaCl). The destabilizing effect on T3 binding induced by exogenous dephosphorylation is more efficiently reversed by β-mercaptoethanol when the extracts were pretreated by molybdate. In controls, the loss of saturable T3 binding activity was recovered by 50% at a 10 mM concentration of β-mercaptoethanol, but in the presence of molybdate, the loss of T3 binding activity was recovered by 50% at a 5 mM concentration of β-mercaptoethanol. This molybdate–receptor interaction is similar to that with nuclear receptor models in term of (i) stabilization of hormone binding, (ii) dependency on a thiol, and (iii) reversibility of the destabilizing effect by exogenous dephosphorylation.Key words: thyroid hormone receptor, binding activity, sodium molybdate, alkaline phosphatase.

scholarly journals Positive and negative modulation of Jun action by thyroid hormone receptor at a unique AP1 site.

1993 ◽  
Vol 13 (5) ◽  
pp. 3042-3049 ◽  
Author(s):  
G Lopez ◽  
F Schaufele ◽  
P Webb ◽  
J M Holloway ◽  
J D Baxter ◽  
...  
Keyword(s):  
Thyroid Hormone ◽  
Transcriptional Activation ◽  
Hormone Receptor ◽  
Thyroid Hormone Receptor ◽  
Binding Activity ◽  
Binding Domain ◽  
Dna Binding Domain ◽  
Plasmid Sequence ◽  
Regulatory Effects

We have characterized the putative AP1 site in the backbone of pUC plasmids and found unique regulatory effects. The site, which mapped to a 19-bp region around nucleotide 37, conferred transcriptional activation by Jun or Jun/Fos that was boosted up to fivefold by unliganded thyroid hormone receptor (TR). Thyroid hormone changed potentiation of the Jun response by TR into repression. Although the plasmid sequence is a near-perfect consensus AP1 site, the perfect consensus AP1 site from the human collagenase promoter did not show the same effects. Deletion of the ligand binding domain of the TR eliminated the ability of the receptor to boost Jun activity, and deletion, mutation, or changes in specificity of the DNA binding domain eliminated both its ability to potentiate Jun activity and repress with hormone. In vitro Jun/Fos complexes bound the operative plasmid fragment, and the presence of TR interfered very little with Jun/Fos binding activity. Protein interaction studies in the absence of DNA showed that TR bound Jun protein in solution either in the presence or in the absence of hormone. These observations suggest a mechanism for synergy and repression by TR through modulation of Jun activity: positive when TR is unliganded, and negative when hormone is bound. They also suggest that the presence of the plasmid element can confound studies of the regulation of linked promoters.

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