scholarly journals Cytosolic glucocorticoid receptor interaction with nuclear factor-kappaB proteins in rat liver cells

2003 ◽  
Vol 373 (1) ◽  
pp. 211-220 ◽  
Author(s):  
Christina WIDÉN ◽  
Jan-Åke GUSTAFSSON ◽  
Ann-Charlotte WIKSTRÖM
Keyword(s):  
Glucocorticoid Receptor ◽  
Western Blotting ◽  
Rat Liver ◽  
Intracellular Localization ◽  
Receptor Interaction ◽  
Physical Interaction ◽  
Inflammatory Factor ◽  
Nuclear Factor

The glucocorticoid receptor (GR) acts as an anti-inflammatory factor. To a large extent, this activity is exerted by the interference of pro-inflammatory nuclear factor κB (NF-κB) activity. In their respective inactive forms, both GR and NF-κB reside in the cytoplasm and translocate to the nucleus on relevant stimulation. Previously, p65, a component of the NF-κB complex, and GR have been shown to interact physically in vitro, and the interaction is assumed to take place in the nucleus of cells [McKay and Cidlowski (1999) Endocrine Rev. 20, 435–459]. We have studied the interaction between GR and NF-κB using in vivo-like conditions. Using immunoaffinity chromatography or immunoprecipitation, combined with Western blotting, we observed that, with endogenous protein levels in cytosolic extracts of rat liver and of H4-II-E-C3 hepatoma cells and in contrast with the current belief, p65, p50 and inhibitory κBα complex interact with GR, even in the absence of glucocorticoid or an inflammatory signal. The interaction between non-liganded/non-activated GR and p65/p50 has also been verified by both p65 and p50 co-immunoprecipitations. Intracellular localization studies, using Western blotting, revealed that glucocorticoids can decrease tumour necrosis factor α (TNFα)-induced nuclear entry of p65, whereas glucocorticoid-induced GR translocation was much less affected by TNFα. We were also able to demonstrate a nuclear interaction of GR and p65 and p50 using in vivo-like protein concentrations. Furthermore, nuclear GR interaction with heat-shock protein 90 was enhanced distinctly by TNFα treatment. In conclusion, our studies suggest a strong interconnectivity between the NF-κB and GR-signalling pathways where also, somewhat unexpectedly, a physical interaction in the cytosol constitutes an integral part of GR–NF-κB cross-talk.

The glucocorticoid receptor binds to a sequence overlapping the TATA box of the human osteocalcin promoter: a potential mechanism for negative regulation

1991 ◽  
Vol 11 (6) ◽  
pp. 3379-3383
Author(s):  
P E Strömstedt ◽  
L Poellinger ◽  
J A Gustafsson ◽  
J Carlstedt-Duke
Keyword(s):  
Glucocorticoid Receptor ◽  
Specific Binding ◽  
Tata Box ◽  
Receptor Interaction ◽  
Receptor Binding Site ◽  
Osteocalcin Gene ◽  
Close Proximity ◽  
Human Osteocalcin

Expression of the human osteocalcin promoter is negatively regulated by glucocorticoids in vivo. In vitro DNase I and exonuclease III footprinting analysis showed binding of purified glucocorticoid receptor in close proximity to and overlapping with the TATA box of the osteocalcin gene. These results imply competition or interference with binding of the TATA box-binding transcription factor IID as a mechanism of repression of this gene by glucocorticoids. In support of this notion, point mutation analysis of the receptor binding site indicated that flanking nucleotides and not the TATA box motif per se were important for receptor interaction. Moreover, DNA binding competition assays showed specific binding of the receptor only to the TATA box region of the osteocalcin gene and not to the corresponding region of an immunoglobulin heavy-chain promoter.

Import of the glucocorticoid receptor into rat liver mitochondria in vivo and in vitro

1993 ◽  
Vol 46 (3) ◽  
pp. 401-413 ◽  
Author(s):  
C. Demonacos ◽  
N.C. Tsawdaroglou ◽  
R. Djordjevic-Markovic ◽  
M. Papalopoulou ◽  
V. Galanopoulos ◽  
...  
Keyword(s):  
Glucocorticoid Receptor ◽  
Rat Liver ◽  
Liver Mitochondria ◽  
Rat Liver Mitochondria

scholarly journals Competitive Cofactor Recruitment by Orphan Receptor Hepatocyte Nuclear Factor 4α1: Modulation by the F Domain

2002 ◽  
Vol 22 (6) ◽  
pp. 1626-1638 ◽  
Author(s):  
Michael D. Ruse ◽  
Martin L. Privalsky ◽  
Frances M. Sladek
Keyword(s):  
Direct Interaction ◽  
Orphan Receptor ◽  
Receptor Interaction ◽  
Dependent Manner ◽  
Nuclear Factor ◽  
Hepatocyte Nuclear Factor ◽  
Interaction Domain ◽  
The Status

ABSTRACT For most ligand-dependent nuclear receptors, the status of endogenous ligand modulates the relative affinities for corepressor and coactivator complexes. It is less clear what parameters modulate the switch between corepressor and coactivator for the orphan receptors. Our previous work demonstrated that hepatocyte nuclear factor 4α1 (HNF4α1, NR2A1) interacts with the p160 coactivator GRIP1 and the cointegrators CBP and p300 in the absence of exogenously added ligand and that removal of the F domain enhances these interactions. Here, we utilized transient-transfection analysis to demonstrate repression of HNF4α1 activity by the corepressor silencing mediator of retinoid and thyroid receptors (SMRT) in several cell lines and on several HNF4α-responsive promoter elements. Glutathione S-transferase pulldown assays confirmed a direct interaction between HNF4α1 and receptor interaction domain 2 of SMRT. Loss of the F domain resulted in marked reduction of the ability of SMRT to interact with HNF4α1 in vitro and repress HNF4α1 activity in vivo, although the isolated F domain itself failed to interact with SMRT. Surprisingly, loss of both the A/B and F domains restored full repression by SMRT, suggesting involvement of both domains in the SMRT interaction. Finally, we show that when coexpressed along with HNF4α1 and GRIP1, CBP, or p300, SMRT can titer out HNF4α1-mediated transactivation in a dose-dependent manner and that this competition derives from mutually exclusive binding. Collectively, these results suggest that HNF4α can functionally interact with both a coactivator and a corepressor without altering the status of any putative ligand and that the presence of the F domain may play a role in discriminating between the different coregulators.

scholarly journals Cellular distribution of nuclear factor κB binding activity in rat liver

1992 ◽  
Vol 287 (2) ◽  
pp. 645-649 ◽  
Author(s):  
A R Freedman ◽  
R J Sharma ◽  
G J Nabel ◽  
S G Emerson ◽  
G E Griffin
Keyword(s):  
Rat Liver ◽  
Kupffer Cells ◽  
Cellular Localization ◽  
Rat Hepatocytes ◽  
Binding Activity ◽  
Nuclear Factor ◽  
Nf Kappa B ◽  
Mobility Shift

The cellular localization of nuclear factor kappa B (NF-kappa B) binding activity in rat liver has been investigated using electrophoretic mobility shift assay on extracts of highly purified hepatocytes and Kupffer cells obtained from liver perfused in vivo with collagenase. Constitutive NF-kappa B binding activity was demonstrated in nuclear extracts of control Kupffer cells, and this was not apparently influenced by injection of lipopolysaccharide (LPS) into rats 24 h before perfusion. In contrast, little nuclear NF-kappa B binding activity was present in hepatocytes from control animals, although there was detectable inactive, inhibitor-bound, NF-kappa B in the cytoplasm. However, nuclear NF-kappa B binding activity was increased in hepatocytes from LPS-treated animals and after in vitro culture of control rat hepatocytes. Thus NF-kappa B binding activity has been demonstrated in highly purified hepatocytes and appears to be inducible both in vivo and in vitro. These findings support a role for NF-kappa B in hepatocyte gene regulation which may be important in the modulation of the hepatic acute phase response.

scholarly journals The glucocorticoid receptor binds to a sequence overlapping the TATA box of the human osteocalcin promoter: a potential mechanism for negative regulation.

1991 ◽  
Vol 11 (6) ◽  
pp. 3379-3383 ◽  
Author(s):  
P E Strömstedt ◽  
L Poellinger ◽  
J A Gustafsson ◽  
J Carlstedt-Duke
Keyword(s):  
Glucocorticoid Receptor ◽  
Specific Binding ◽  
Tata Box ◽  
Receptor Interaction ◽  
Receptor Binding Site ◽  
Osteocalcin Gene ◽  
Close Proximity ◽  
Human Osteocalcin

Expression of the human osteocalcin promoter is negatively regulated by glucocorticoids in vivo. In vitro DNase I and exonuclease III footprinting analysis showed binding of purified glucocorticoid receptor in close proximity to and overlapping with the TATA box of the osteocalcin gene. These results imply competition or interference with binding of the TATA box-binding transcription factor IID as a mechanism of repression of this gene by glucocorticoids. In support of this notion, point mutation analysis of the receptor binding site indicated that flanking nucleotides and not the TATA box motif per se were important for receptor interaction. Moreover, DNA binding competition assays showed specific binding of the receptor only to the TATA box region of the osteocalcin gene and not to the corresponding region of an immunoglobulin heavy-chain promoter.

Glucocorticoid receptor-mediated alleviation of inflammation by berberine: in vitro, in silico and in vivo investigations

2021 ◽  
Author(s):  
Yuan Liang ◽  
Tiehua Zhang ◽  
Jingqi Zhao ◽  
Chenfei Li ◽  
Haoyang Zou ◽  
...  
Keyword(s):  
Glucocorticoid Receptor ◽  
In Silico ◽  
Biological Activities ◽  
Inflammatory Factor ◽  
Anti Inflammatory ◽  
Inflammatory Effect

As a natural dietary ingredient, berberine possesses multiple biological activities including anti-inflammatory effect. Specifically, berberine can inhibit the lipopolysaccharide-induced mRNA expressions of inflammatory factor. In this work, glucocorticoid receptor (GR)-mediated...

Effect of X-irradiation on gene expression of rat liver chemokines: in vivo and in vitro studies

2008 ◽  
Vol 46 (01) ◽  
Author(s):  
F Moriconi ◽  
H Christiansen ◽  
H Christiansen ◽  
N Sheikh ◽  
J Dudas ◽  
...  
Keyword(s):  
Gene Expression ◽  
Rat Liver ◽  
In Vitro Studies ◽  
X Irradiation

L-Tetrahydropalmatine Inhibits the Progression of Glioblastoma Tumor by Suppressing the Extracellular-Signal-Regulated Kinase/Nuclear Factor-Kappa B Signaling Pathway

2020 ◽  
Vol 19 (2) ◽  
pp. 164-171
Author(s):  
Feng Xue ◽  
Tingting Chen
Keyword(s):  
Glioblastoma Multiforme ◽  
Nuclear Factor Kappa B ◽  
Matrix Metalloproteinase 2 ◽  
Nuclear Factor ◽  
Nuclear Factor Kappa ◽  
Extracellular Signal ◽  
Apoptosis Protein ◽  
Endothelial Growth Factor

Glioblastoma multiforme is the most common malignancy of central nervous system. Herein we have evaluated the effect of L-tetrahydropalmatine, an isoquinoline alkaloid, on the tumor growth both in vivo and in vitro using C6 glioblastoma multiforme cells and BALB/c mice injected subcutaneously with C6/luc2 cells. The results of these studies show that L-tetrahydropalmatine exhibited cytotoxic effect on C6 glioblastoma multiforme cells, suppressed nuclear factor-kappa B activity, suppressed the levels of tumor-linked proteins such as matrix metalloproteinase-2/9, Cyclin-D1, vascular endothelial growth factor, and X-linked inhibitor of apoptosis protein via ERK/nuclear factor-kappa B cascade. Further, L-tetrahydropalmatine inhibited the cell migration and invasion properties of C6 cells, and also suppressed the tumor weight and volume in mice. Immunohistochemical staining of tumor tissues suggested that L-tetrahydropalmatine inhibited the extracellular-signal-regulated kinase/nuclear factor-kappa B cascade and suppressed the levels of Cyclin-D1; matrix metalloproteinase-2/9; X-linked inhibitor of apoptosis protein; and vascular endothelial growth factor, and also the progression and growth of glioblastoma multiforme in mice. In summary, L-tetrahydropalmatine inhibits the ERK/nuclear factor-kappa B cascade, decreases the tumor volume, and inhibits the proteins responsible for tumor growth both in vivo and in vitro.

Sign in / Sign up

Export Citation Format

Share Document