Antibodies specific to the retinoic acid human nuclear receptors alpha and beta.

Gaub, Marie–Pierre; Lutz, Yves; Ruberte, Esther; Petkovich, Martin; Brand, Nigel J.; Chambon, Pierre

doi:10.1073/pnas.86.9.3089

Cited by 62 publications

(22 citation statements)

References 15 publications

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“…The lack of factor occupancy seen prior to RA treatment is not due to the cytoplasmic partitioning of the receptors, as exemplified by the glucocorticoid hormone receptor (GR). Much of the GR is present in the cytoplasm before hormone stimulation, but the GR is translocated into the nucleus following ligand administration (48 prior to ligand addition (21,36). RA induction of factor occupancy is unlikely to be due to increased receptor concentrations in P19 cells, since there was no significant change in the levels of PRARE-binding activity during the initial 4 h of RA treatment (Fig.…”

Section: Discussionmentioning

confidence: 99%

Ligand-dependent occupancy of the retinoic acid receptor beta 2 promoter in vivo.

Dey¹,

Minucci²,

Ozato³

1994

Mol. Cell. Biol.

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Retinoic acid (RA) activates transcription of the RA receptor 02 (RAR02) gene in embryonal carcinoma (EC) cells. This activation involves binding of the RAR/retinoid X receptor (RAR/RXR) heterodimer to the RA-responsive element (PRARE). Dimethyl sulfate-based genomic footprinting was performed to examine occupancy of this promoter in P19 EC cells. No footprint was detected at the PRARE prior to RA treatment, but a footprint was detected within the first hour of RA treatment. Concomitantly, other elements in the promoter, the cyclic AMP-responsive element and tetradecanoyl phorbol acetate-like-responsive element became footprinted. Footprints at these elements were induced by RA without requiring new protein synthesis and remained for the entire duration of RA treatment but rapidly reversed upon withdrawal of RA. A delayed protection observed at the initiator site was also reversed upon RA withdrawal. The RA-inducible footprint was not due to induction of factors that bind to these element, since in vitro assays showed that these factors are present in P19 cell extracts before RA treatment. Significantly, no RA-induced footprint was observed at any of these elements in P19 cells expressing a dominant negative RXR0, in which RXR heterodimers are unable to bind to the PIRARE. Results indicate that binding of a liganded heterodimer receptor to the PRARE is the initial event that allows other elements to gain access to the factors. In accordance, reporter analyses showed that a mutation in the PRARE, but not those in other elements, abrogates RA activation of the promoter. It is likely that the RAR02 promoter opens in a hierarchically ordered manner, signalled by the occupancy of liganded heterodimers.

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Section: Discussionmentioning

confidence: 99%

Ligand-dependent occupancy of the retinoic acid receptor beta 2 promoter in vivo.

Dey¹,

Minucci²,

Ozato³

1994

Mol. Cell. Biol.

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“…The receptorlike specificity found in the competition experiments, however, strongly supports the specific detection of MR. It is also possible that, as with thyroid (23) and retinoic acid receptors (24,25), multiple forms of MR exist, which have different physiological functions requiring diverse partitioning in cell compartments. Two forms of receptors differing by their intracellular distribution and functions have already been suggested for the glucocorticoid receptor (19).…”

Section: Resultsmentioning

confidence: 99%

Immunohistochemical localization of renal mineralocorticoid receptor by using an anti-idiotypic antibody that is an internal image of aldosterone.

Lombès

Farman

Oblin

et al. 1990

Proc. Natl. Acad. Sci. U.S.A.

118

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A monoclonal antibody (H1OE), generated by an auto-anti-idiotypic procedure and directed at the aldosterone-binding site of mineralocorticoid receptor (MR), was used in immunohistochemical studies to localize MR in rabbit kidney preparations. In agreement with earlier physiological and biochemical observations, MR was detected in connecting and cortical collecting tubules. Additionally, MR was detected in the distal tubules, the medullary and papillary collecting ducts, and in the epithelial cells lining the papilla. MATERIALS AND METHODSReagents. Aldosterone, dexamethasone, and estradiol were from Sigma. Deoxycorticosterone acetate and RU 486 were from Roussel-Uclaf (Romainville, France). SC 9420 was from Searle.Tissue Preparation. New Zealand White female rabbits weighing 1.5 kg were used in this study. Bilateral adrenalectomy was performed under Fluothan anesthesia. The animals received Syncortyl (1.9 mg/kg; deoxycorticosterone acetate) i.p. at the time of operation and the 2 following days and were given 0.9% saline to drink ad libitum. These animals were then used 5 to 7 days after surgery. Aldosterone-treated rabbits received an i.p. injection of 1.5 mg of aldosterone 45 min before the study.Normal, adrenalectomized, or aldosterone-treated rabbits were anesthetized with 25 to 50 mg of Nembutal and perfused through the abdominal aorta with 500 ml of Zamboni's solution [2% (wt/vol) paraformaldehyde/15% (vol/vol) saturated picric acid solution/300 mM (85%, vol/vol) sodium phosphate buffer, pH 7.4] at 37°C. The perfusion procedure was done as described (3). At the end of perfusion, the kidneys were removed, cut into slices and pyramids (-2 mm thick), and postfixed for 24 hr in the Zamboni fixative. The kidney slices and pyramids were washed in 70o (vol/vol) ethanol, dehydrated in graded ethanol, cleared in 1-butanol, and embedded in Paraplast. Sections (7 ,um) were cut, mounted on histological slides, and processed for immunohistochemistry.Immunohistochemical Technique. A routine procedure of indirect immunostaining was used (4). Briefly, after deparaffinization and rehydration, sections were incubated with 3% normal horse serum in phosphate-buffered saline (PBS) solution. The anti-idiotypic mAb H1OE, a mouse IgG1 immunoglobulin, was used as diluted ascites fluid (1). A nonimmune preparation of mouse IgG (prepared in the laboratory) was used as control. The mAb was used at -1-5 ,g/ml, followed by a horse biotinylated anti-mouse IgG antibody (Vector Laboratories). The avidin-biotin-peroxidase complex (ABC-Elite from Vector Laboratories) was used as a detection system. After each incubation step, the slides were rinsed in PBS. Peroxidase activity was revealed by diamiAbbreviations: MR, mineralocorticoid receptor; mAb, monoclonal antibody.§To whom reprint requests should be addressed. 1086The publication costs of this article were defrayed in part by page charge payment. This article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. §1734 solely to indicate this fact.

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“…This hypothesis was based on transactivation experiments, showing a dominant negative activity of PML/RARa on RARa (Kakizuka et al, 1991;De TheÂ et al, 1991;Pandol® et al, 1991;Kastner et al, 1992), and on the nuclear localization pattern of these proteins. RARa is nuclear di use (Gaub et al, 1989), PML is localized within speci®c subnuclear structures called PML nuclear bodies, while PML/RARa has distinct distribution pattern (microspeckled localization) (Dyck et al, 1994;Weis et al, 1994;Koken et al, 1994;Flenghi et al, 1995). Expression of PML/RARa produces delocalization within microspeckles of PML and RXR, an RARa cofactor (Chambon, 1996), and so might alter the function of both PML and RARa.…”

Section: Introductionmentioning

confidence: 99%

Formation of PML/RARα high molecular weight nuclear complexes through the PML coiled-coil region is essential for the PML/RARα-mediated retinoic acid response

et al. 1999

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Antibodies specific to the retinoic acid human nuclear receptors alpha and beta.

Cited by 62 publications

References 15 publications

Ligand-dependent occupancy of the retinoic acid receptor beta 2 promoter in vivo.

Ligand-dependent occupancy of the retinoic acid receptor beta 2 promoter in vivo.

Immunohistochemical localization of renal mineralocorticoid receptor by using an anti-idiotypic antibody that is an internal image of aldosterone.

Formation of PML/RARα high molecular weight nuclear complexes through the PML coiled-coil region is essential for the PML/RARα-mediated retinoic acid response

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