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  • 1
    Electronic Resource
    Electronic Resource
    Springer
    Molecular and cellular biochemistry 194 (1999), S. 199-207 
    ISSN: 1573-4919
    Keywords: steroid receptor superfamily ; tetracycline inducible system ; differential display ; TR2 orphan receptor
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology , Medicine
    Notes: Abstract We have identified a DNA response element (TR2RE-HR) in the 3′ flanking region of the human histamine H1 receptor gene as a target for the TR2 orphan receptor, a member of the steroid/thyroid hormone receptor superfamily. The application of both tetracycline inducible and improved differential display systems has allowed us to isolate a cDNA fragment differentially regulated by the expression of the TR2 orphan receptor. Northern blot and sequencing analysis demonstrated that the expression of the human histamine H1 receptor gene was differentially repressed by the TR2 orphan receptor. Electrophoretic mobility shift assay further revealed a specific binding (dissociation constant = 26.2 nM) between the TR2 orphan receptor and the wildtype TR2RE-HR, but not the mutant TR2RE-HR. In addition, reporter gene expression assay indicated that the TR2 orphan receptor may suppress the expression of luciferase activities in a dose-dependent manner via the TR2RE-HR in HeLa cells. Our results demonstrate that the histamine H1 receptor gene could represent one of the target genes directly regulated by the human TR2 orphan receptor.
    Type of Medium: Electronic Resource
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  • 2
    ISSN: 1520-4995
    Source: ACS Legacy Archives
    Topics: Biology , Chemistry and Pharmacology
    Type of Medium: Electronic Resource
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  • 3
    ISSN: 1546-170X
    Source: Nature Archives 1869 - 2009
    Topics: Biology , Medicine
    Notes: [Auszug] Motor neuron degeneration resulting from the aggregation of the androgen receptor with an expanded polyglutamine tract (AR-polyQ) has been linked to the development of spinal and bulbar muscular atrophy (SBMA or Kennedy disease). Here we report that adding ...
    Type of Medium: Electronic Resource
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  • 4
    ISSN: 1573-4919
    Keywords: TR4 orphan receptor ; estrogen receptor ; thyroid hormone receptor
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology , Medicine
    Notes: Abstract We demonstrate that TR2 orphan receptor (TR2) may induce transactivation activities via an AGGTCA-like-direct-repeat-4 consensus thyroid hormone response element (DR4-TRE) system. TR2 showed a slightly greater binding affinity than thyroid hormone receptor α1 (TRα1)/retinoid X receptor α (RXRα) heterodimer with Kds 0.5 nM and 2.3 nM, respectively. These receptors, TR2 and TRα1/RXRα heterodimer, competed with each other on binding to limited amounts of DR4-TRE. TR2 canceled the suppression effect of unliganded-TRα1 on CAT reporter activity in a dose-dependent fashion. Estrogen receptor (ER) and 2P2 (a mutated TR2 with P box sequence of androgen receptor) failed not only to bind to DR4-TRE but also to recover this inhibitory effect of unliganded TRα1. However, when T3 was supplemented, estradiol-ER competed for a full CAT activity while TR2 showed an additive effect on the transcriptional activation. These results indicate that DNA binding is essential for TR2 to take action and fully functional liganded TRa1 may rely on common factors shared with ER but not TR2.
    Type of Medium: Electronic Resource
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  • 5
    ISSN: 1573-4919
    Keywords: testosterone ; DHT ; AR ; ARE
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology , Medicine
    Notes: Abstract There are two major physiological androgens, testosterone (T), and 5α-dihydrotestosterone (DHT), which induce different responses in mammals. These androgens regulate the target gene transcription via binding to and activating the same androgen receptor (AR). The molecular mechanisms that differ between these two very close androgens through the same AR protein to target the distinct genomic responses remain unknown. Using yeast genetic selection, we identified two kinds of androgen response elements (ARE), which could respond differentially to T vs. DHT. These two AREs also show different T- vs. DHT-induced AR transactivation in mammalian Chinese hamster ovary (CHO) cells in terms of copy number and comparisons with the classic mouse mammary tumor virus ARE. Together, our results suggest that the selective ARE sequence may play an important role in the differential T- vs. DHT-induced AR transactivation.
    Type of Medium: Electronic Resource
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  • 6
    ISSN: 1573-4919
    Keywords: flow cytometry ; immunohistochemical staining ; ELISA ; prostate cancer ; monoclonal antibody
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology , Medicine
    Notes: Abstract Several monoclonal antibodies (mAbs) and novel mAb-based assays for the androgen receptors (AR) have been developed. Large amounts of the recombinant human AR protein produced by a baculovirus expression system were used as an antigen to produce mAbs. Twenty-nine AR-specific mAbs were first confirmed by Western blot analysis and were then characterized for their immunoglobulin isotypes, epitopes, and epitope localization in AR. Novel assays using flow cytometry and sandwich enzyme-linked immunosorbent assays (ELISA) were established to detect AR-expressing cells and to quantify soluble AR protein, respectively. Using immunostaining, we identified several anti-AR mAbs exclusively recognizing AR within the nuclei of the prostate cancer cell line LNCaP and of prostate tissues in both frozen and paraffin-embedded sections, whereas other mAbs could detect AR in both nuclear and cytoplasmic compartments. Interestingly, certain mAbs, such as G122-25 and G122-77, could distinguish the androgen-bound AR from the unoccupied AR. In sum, many purified AR protein and anti-AR mAbs, together with the assays developed, could be powerful tools for the study of functional AR and for the diagnosis of prostatic cancers.
    Type of Medium: Electronic Resource
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  • 7
    Electronic Resource
    Electronic Resource
    Springer
    Molecular and cellular biochemistry 195 (1999), S. 19-23 
    ISSN: 1573-4919
    Keywords: androgen receptor ; baculovirus expression system ; androgen response element
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology , Medicine
    Notes: Abstract To purify the androgen receptor (AR) efficiently from baculovirus expression system, we fused 6 histidine residues with the N-terminal domain of AR as a tag to specifically bind to Ni+2-affinity column. Our data indicated that adding androgen can increase the binding capacity of his-tag AR to the Ni+2-affinity column, and this increased binding capacity of AR could be due to the exposure of histidine residues of N-terminal domain induced by androgen. The androgen-enhanced binding to Ni+2-column also correlated with the increasing solubility of AR. Electrophoretic mobility shift assay further indicated that only purified AR could interact with androgen response element. Together, our data suggest that the binding of androgen to the hormone binding domain of AR may result in the conformational change of the N-terminal domain of AR and increase the hydrophilic property of AR.
    Type of Medium: Electronic Resource
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