scholarly journals Fluorescence Anisotropy Microplate Assay to Investigate the Interaction of Full-Length Steroid Receptor Coactivator-1a with Steroid Receptors

2013 ◽  
pp. 339-351 ◽  
Author(s):  
Chen Zhang ◽  
Steven K. Nordeen ◽  
David J. Shapiro
Keyword(s):  
Fluorescence Anisotropy ◽  
Steroid Receptors ◽  
Steroid Receptor ◽  
Full Length ◽  
Microplate Assay ◽  
Steroid Receptor Coactivator

scholarly journals Reduction of Coactivator Expression by Antisense Oligodeoxynucleotides Inhibits ERα Transcriptional Activity and MCF-7 Proliferation

2002 ◽  
Vol 16 (2) ◽  
pp. 253-270 ◽  
Author(s):  
Ilaria T. R. Cavarretta ◽  
Ratna Mukopadhyay ◽  
David M. Lonard ◽  
Lex M. Cowsert ◽  
C. Frank Bennett ◽  
...  
Keyword(s):  
Gene Expression ◽  
Transcriptional Activity ◽  
Steroid Receptors ◽  
Biological Effects ◽  
Steroid Receptor ◽  
Transient Transfection ◽  
Antisense Oligodeoxynucleotides ◽  
Steroid Receptor Coactivator ◽  
Mcf 7

Abstract Steroid receptor RNA activator (SRA) is a novel coactivator for steroid receptors that acts as an RNA molecule, whereas steroid receptor coactivator (SRC) family members, such as steroid receptor coactivator-1 (SRC-1) and transcriptional intermediary factor 2 (TIF2) exert their biological effects as proteins. Individual overexpression of each of these coactivators, which can form multimeric complexes in vivo, results in stimulated ERα transcriptional activity in transient transfection assays. However there is no information on the consequences of reducing SRC-1, TIF2, or SRA expression, singly or in combination, on ERα transcriptional activity. We therefore developed antisense oligodeoxynucleotides (asODNs) to SRA, SRC-1, and TIF2 mRNAs, which rapidly and specifically reduced the expression of each of these coactivators. ERα-dependent gene expression was reduced in a dose-dependent fashion by up to 80% in cells transfected with these oligonucleotides. Furthermore, treatment of cells with combinations of SRA, SRC-1, and TIF2 asODNs reduced ERα transcriptional activity to an extent greater than individual asODN treatment alone, suggesting that these coactivators cooperate, in at least an additive fashion, to activate ERα-dependent target gene expression. Finally, treatment of MCF-7 cells with asODN against SRC-1 and TIF2 revealed a requirement of these coactivators, but not SRA, for hormone-dependent DNA synthesis and induction of estrogen-dependent pS2 gene expression, indicating that SRA and SRC family coactivators can fulfill specific functional roles. Taken together, we have developed a rapid method to reduce endogenous coactivator expression that enables an assessment of the in vivo role of specific coactivators on ERα biological action and avoids potential artifacts arising from overexpression of coactivators in transient transfection assays.

scholarly journals Fluorescence anisotropy microplate assay for analysis of steroid receptor-DNA interactions

BioTechniques ◽  
2004 ◽  
Vol 37 (5) ◽  
pp. 807-817 ◽  
Author(s):  
Stanley Y. Wang ◽  
Bonnie S. Ahn ◽  
Rebecca Harris ◽  
Steven K. Nordeen ◽  
David J. Shapiro
Keyword(s):  
Fluorescence Anisotropy ◽  
Steroid Receptor ◽  
Microplate Assay ◽  
Dna Interactions

scholarly journals Steroid receptor coactivator-2 expression in brain and physical associations with steroid receptors

Neuroscience ◽  
2010 ◽  
Vol 169 (3) ◽  
pp. 1017-1028 ◽  
Author(s):  
M.A. Yore ◽  
D. Im ◽  
L.K. Webb ◽  
Y. Zhao ◽  
J.G. Chadwick ◽  
...  
Keyword(s):  
Steroid Receptors ◽  
Steroid Receptor ◽  
Steroid Receptor Coactivator

scholarly journals Transcriptional activation by estrogen receptor (ERalpha) and steroid receptor coactivator (SRC1) involves distinct mechanisms in yeast and mammalian cells

2003 ◽  
Vol 30 (3) ◽  
pp. 411-422 ◽  
Author(s):  
HM Sheppard ◽  
S Matsuda ◽  
JC Harries ◽  
KB Kindle ◽  
DM Heery
Keyword(s):  
Transcriptional Activation ◽  
Mammalian Cells ◽  
Transcriptional Activity ◽  
Steroid Receptors ◽  
Steroid Receptor ◽  
Yeast Cells ◽  
Creb Binding Protein ◽  
Subtle Difference ◽  
Steroid Receptor Coactivator ◽  
Helix 12

Steroid receptors activate transcription in yeast cells via interactions with endogenous coactivators and/or basal factors. We examined the effects of mutations in the ligand binding domain on the transcriptional activity of ERalpha in yeast. Our results show that mutations in Helix 3 (K366A) and Helix 12 (M547A, L548A) disrupt transcriptional activity of ERalpha in yeast, as previously observed in mammalian cells. However, replacement of a conserved tyrosine residue in Helix 12 with alanine or aspartate (Y541A and Y541D), which renders ERalpha constitutively active in mammalian cells, had only a weak stimulatory effect on ligand-independent reporter activation by ERalpha in yeast. Two-hybrid interaction experiments revealed that a Y541A mutant expressed in yeast was capable of ligand-independent binding to a mammalian coactivator, suggesting that there is a subtle difference in how this mutant interacts with mammalian and yeast cofactors. We also show that the ligand-dependent activities of ERalpha and progesterone receptor (PR) in yeast cells were strongly enhanced by the human p160 protein steroid receptor coactivator (SRC1), but not by CREB-Binding Protein (CBP) or the p300/CBP associated factor (P/CAF). Although the SRC1 activation domains AD1 and AD2 are functional in yeast, deletion of these sequences only partially impaired SRC1 coactivator function in this organism; this is in contrast to similar experiments in mammalian cells. Thus SRC1 sequences involved in recruitment of CBP/p300 and Co-Activator-Associated Arginine Methyltransferase (CARM-1) in mammalian cells are not essential for its function in yeast, suggesting that SRC1 operates via distinct mechanisms in yeast and mammalian cells.

scholarly journals Peptidyl-Prolyl Isomerase 1 (Pin1) Serves as a Coactivator of Steroid Receptor by Regulating the Activity of Phosphorylated Steroid Receptor Coactivator 3 (SRC-3/AIB1)

2005 ◽  
Vol 25 (21) ◽  
pp. 9687-9699 ◽  
Author(s):  
Ping Yi ◽  
Ray-Chang Wu ◽  
Joshua Sandquist ◽  
Jiemin Wong ◽  
Sophia Y. Tsai ◽  
...  
Keyword(s):  
Conformational Changes ◽  
Intracellular Signaling ◽  
Steroid Receptors ◽  
Steroid Receptor ◽  
Dependent Manner ◽  
Prolyl Isomerase ◽  
Peptidyl Prolyl Isomerase ◽  
Steroid Receptor Coactivator

ABSTRACT Steroid receptor coactivator 3 (SRC-3/AIB1) interacts with steroid receptors in a ligand-dependent manner to activate receptor-mediated transcription. A number of intracellular signaling pathways initiated by growth factors and hormones induce phosphorylation of SRC-3, regulating its function and contributing to its oncogenic potential. However, the range of mechanisms by which phosphorylation affects coactivator function remains largely undefined. We demonstrate here that peptidyl-prolyl isomerase 1 (Pin1), which catalyzes the isomerization of phosphorylated Ser/Thr-Pro peptide bonds to induce conformational changes of its target proteins, interacts selectively with phosphorylated SRC-3. In addition, Pin1 and SRC-3 activate nuclear-receptor-regulated transcription synergistically. Depletion of Pin1 by small interfering RNA (siRNA) reduces hormone-dependent transcription from both transfected reporters and an endogenous steroid receptor target gene. We present evidence that Pin1 modulates interactions between SRC-3 and CBP/p300. The interaction is enhanced in vitro and in vivo by Pin1 and diminished when cellular Pin1 is reduced by siRNA or in stable Pin1-depleted cell lines. Depletion of Pin1 in MCF-7 human breast cancer cells reduces the endogenous estrogen-dependent recruitment of p300 to the promoters of estrogen receptor-dependent genes. Pin1 overexpression enhanced SRC-3 cellular turnover, and depletion of Pin1 stabilized SRC-3. Our results suggest that Pin1 functions as a transcriptional coactivator of nuclear receptors by modulating SRC-3 coactivator protein-protein complex formation and ultimately by also promoting the turnover of the activated SRC-3 oncoprotein.

scholarly journals Steroid Receptor Coactivator 3 Regulates Synaptic Plasticity and Hippocampus-dependent Memory

2021 ◽  
Author(s):  
Hai-Long Zhang ◽  
Bing Zhao ◽  
Pin Yang ◽  
Yin-Quan Du ◽  
Wei Han ◽  
...  
Keyword(s):  
Synaptic Plasticity ◽  
Steroid Hormones ◽  
Hippocampal Neurons ◽  
Short Term Memory ◽  
Steroid Receptors ◽  
Long Term Potentiation ◽  
Steroid Receptor ◽  
Steroid Receptor Coactivator ◽  
Term Potentiation ◽  
And Function

AbstractSteroid hormones play important roles in brain development and function. The signaling of steroid hormones depends on the interaction between steroid receptors and their coactivators. Although the function of steroid receptor coactivators has been extensively studied in other tissues, their functions in the central nervous system are less well investigated. In this study, we addressed the function of steroid receptor coactivator 3 (SRC3) – a member of the p160 SRC protein family that is expressed predominantly in the hippocampus. While hippocampal development was not altered in Src3+/− mice, hippocampus-dependent functions such as short-term memory and spatial memory were impaired. We further demonstrated that the deficient learning and memory in Src3+/− mice was strongly associated with the impairment of long-term potentiation (LTP) at Schaffer Collateral-CA1 synapses. Mechanistic studies indicated that Src3+/− mutation altered the composition of N-methyl-D-aspartate receptor subunits in the postsynaptic densities of hippocampal neurons. Finally, we showed that SRC3 regulated synaptic plasticity and learning mainly dependent on its lysine acetyltransferase activity. Taken together, these results reveal previously unknown functions of SRC3 in the hippocampus and thus may provide insight into how steroid hormones regulate brain function.

Steroid receptor coactivator-1 regulates glioma angiogenesis through polyomavirus enhancer activator 3 signaling

2019 ◽  
Vol 97 (4) ◽  
pp. 488-496
Author(s):  
Yi Zhang ◽  
Wei Shi
Keyword(s):  
Endothelial Cell ◽  
Growth Factor ◽  
Steroid Receptors ◽  
Glioma Cells ◽  
Steroid Receptor ◽  
Angiogenic Factors ◽  
Endothelial Cell Migration ◽  
Steroid Receptor Coactivator

Steroid receptor coactivator 1 (SRC-1) is a transcriptional coactivator for steroid receptors and other transcription factors. SRC-1 has been shown to play an important role in the progression of breast cancer and prostate cancer. However, its role in glioma progression remains unknown. Here, in this study, we report that SRC-1 is upregulated in the vessels of human glioma and exerts important regulatory functions. Specifically, SRC-1 expression significantly enhanced basic fibroblast growth factor (bFGF)-mediated angiogenesis in vivo. Downregulating of SRC-1 expression suppressed endothelial cell migration and tube formation in vitro and upregulated the expression of pro-angiogenic factors, including vascular endothelial growth factor (VEGF) and matrix metallopeptidase (MMP)-9 in glioma cells. These SRC-1-mediated effects were dependent on the activation of polyomavirus enhancer activator 3 (PEA3) transcriptional activity. VEGF and VEGF inducer GS4012 induced the direct binding of SRC-1 and PEA3 in glioma cells, and PEA3 could directly bind with VEGF and MMP-9 promoter under GS4012 treatment in glioma cell. The expression of pro-angiogenic factors induced by SRC-1 was abrogated by sh-PEA3 knockdown. Taken together, these novel outcomes indicated that SRC-1 modulated endothelial cell (EC) function and facilitated a pro-angiogenic microenvironment through PEA3 signaling. Moreover, a combination of targeting SRC-1 and PEA3 signaling in glioma could be a promising strategy for suppressing tumor angiogenesis.

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