Retinoic acid receptor expression vector inhibits differentiation of F9 embryonal carcinoma cells.

Espeseth, Amy S.; Murphy, Shawn P.; Linney, Elwood

doi:10.1101/gad.3.11.1647

Cited by 72 publications

(40 citation statements)

References 48 publications

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“…20; data not shown), further supports the notion that it is the ability to form inactive heterodimers that provides the mutant RARs with their wide spectrum of dominant negative activity. However, this model does not provide an explanation for the apparent difference between our results and those of Espeseth et al (35), who constructed a chimeric RAR-lacZ fusion gene that can function as a dominant repressor. In this mutant, it may be the ability of the lacZ protein to form tetrameric structures that compensates for loss of the authentic dimerization domain.…”

Section: Discussioncontrasting

confidence: 56%

Functional inhibition of retinoic acid response by dominant negative retinoic acid receptor mutants.

Damm

Heyman

Umesono

et al. 1993

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

183

144

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The diverse effects of retinoids on the development, growth, and homeostasis of vertebrate organisms are mediated in part by three distinct isoforms of retinoic acid receptors (RARs). These proteins, which are structurally and functionally closely related to thyroid hormone receptors and the oncogene product v-ErbA, regulate patterns of gene expression in target tissues. One approach to study the distinct effects of retinoic acid in cells is to subvert this activity of endogenous receptors by expression of dominant negative receptor derivatives. We demonstrate here that RARa, RARf8, and RARy can be converted into potent negative transcriptional regulators that block wild-type RAR function. Furthermore, these mutant RARs, but not the wild-type receptors, actively repress the basal transcription level of target promoters. When expressed in transgenic mice, the most potent of these inhibitory receptor mutants is apparently able to disturb developmental processes by inducing a cleft palate in transgenic offspring.Retinoids and, in particular, retinoic acid (RA) are a group of vitamin A-derived substances that exert striking effects on vertebrate development. RA can promote differentiation of a wide variety of cell types in vitro, while in vivo RA has been shown to provide positional information and to influence pattern formation (for review, see refs. 1 and 2). RA is also known to be teratogenic in mammals, causing a range of abnormalities in the face, cleft palate, and malformations of the central nervous system as well as the limbs (3, 4).The identification of three distinct isoforms of RA receptors (RARs; refs. 5-11) as members of the nuclear receptor superfamily provides a model for the molecular mechanism of action of RA. This family of receptors controls cell function by regulating gene expression in target tissues in a ligandinducible fashion. With the exception of the N and C termini, which are almost completely divergent (9), the three RARs are remarkably similar to one another and are structurally and functionally closely related to thyroid hormone receptors (12, 13). Recently, a second RA response pathway has been identified and shown to be mediated by a family of retinoid X receptors (RXRs; refs. 14-17). RXRs differ from RARs in primary sequence and in their response to retinoids (18, 19). They have also been shown to function as coregulators of RARs, thyroid hormone receptors (TRs), and vitamin D3 receptors (VDRs) (16,17,20).We have previously described that the viral homologue of the TR, the v-erbA oncogene product, can function as a potent TR antagonist (21). The constitutive negative phenotype of v-ErbA is due to mutations in the C terminus of the TR (21-23), a region where v-ErbA, the TR, and RARs exhibit 53% amino acid similarity. In the present study, a further characterization of the structure/function relationship of these receptors has allowed us to convert RARa, RAR,3, and RARy into potent negative transcriptional regulators that no longer respond to RA. Furthermore, these mutant RARs ...

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Section: Discussioncontrasting

confidence: 56%

Functional inhibition of retinoic acid response by dominant negative retinoic acid receptor mutants.

Damm

Heyman

Umesono

et al. 1993

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

183

144

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“…It has been shown that RA receptor/retinoid X receptor (RARIRXR) heterodimers bind to these elements in an asymmetrical fashion (9,33,35,39,47,65). The critical involvement of the RAR in RA-induced differentiation in EC cells is supported by the inability of cells expressing a mutated RAR to respond to RA and to undergo differentiation (17,50). A null mutant of RARy also affects RA-induced differentiation (7).…”

mentioning

confidence: 93%

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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“…The ␤-actin-mHSF2-␣ plasmid was constructed by replacing a BglII-HindIII fragment of the HSF2-␤ coding sequence with a corresponding fragment of the HSF2-␣ cDNA. The transfections were performed by using calcium phosphate precipitation as described by Espeseth et al (5). Mouse NIH 3T3 cells were seeded at a density of 500,000 per plate and transfected with 10 g of plasmid DNA per plate the following day.…”

Section: Methodsmentioning

confidence: 99%

Tissue-Dependent Expression of Heat Shock Factor 2 Isoforms with Distinct Transcriptional Activities

Goodson

Park-Sarge

Sarge

1995

Molecular and Cellular Biology

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Heat shock factor 2 (HSF2) functions as a transcriptional regulator of heat shock protein gene expression in mammalian cells undergoing processes of differentiation and development. Our previous studies demonstrated high regulated expression and unusual constitutive DNA-binding activity of the HSF2 protein in mouse testes, suggesting that HSF2 functions to regulate heat shock protein gene expression in spermatogenic cells. The purpose of this study was to test whether HSF2 regulation in testes is associated with alterations in the HSF2 polypeptide expressed in testes relative to other mouse tissues. Our results show that mouse cells express not one but two distinct HSF2 proteins and that the levels of these HSF2 isoforms are regulated in a tissue-dependent manner. The testes express predominantly the 71-kDa HSF2-␣ isoform, while the heart and brain express primarily the 69-kDa HSF2-␤ isoform. These isoforms are generated by alternative splicing of HSF2 pre-mRNA, which results in the inclusion of an 18-amino-acid coding sequence in the HSF2-␣ mRNA that is skipped in the HSF2-␤ mRNA. HSF2 alternative splicing is also developmentally regulated, as our results reveal a switch in expression from the HSF2-␤ mRNA isoform to the HSF2-␣ isoform during testis postnatal development. Transfection analysis shows that the HSF2-␣ protein, the predominant isoform expressed in testis cells, is a more potent transcriptional activator than the HSF2-␤ isoform. These results reveal a new mechanism for the control of HSF2 function in mammalian cells, in which regulated alternative splicing is used to modulate HSF2 transcriptional activity in a tissue-dependent manner.Heat shock factors (HSFs) function as transcriptional regulators of heat shock protein (hsp) gene expression in eukaryotic cells (7,8,10,22). Cells of a number of eukaryotic species contain several related HSF polypeptides, which are expressed from different members of an HSF gene family (12,13,(17)(18)(19). Mouse cells express two different HSF polypeptides, HSF1 and HSF2, which have distinct functional roles in the regulation of hsp gene expression. HSF1 is responsible for mediating the cellular stress response, which is the induction of hsp gene expression that occurs in cells following exposure to elevated temperature and other environmental stress conditions (1, 15). In response to stress conditions, HSF1 undergoes oligomerization from a non-DNA-binding monomeric form to a trimeric, DNA-binding form which interacts with hsp gene promoter sequences to increase transcription of these genes (7,8,10,22).HSF2, on the other hand, appears to function as a regulator of hsp gene expression under nonstress conditions, particularly in cells involved in processes of differentiation and development (11,16,20,21). We have previously shown that both the expression and functional properties of HSF2 are regulated in mouse testis cells (16). HSF2 mRNA expression in testes is subject to developmental, spermatogenesis stage-dependent, and cell-type-dependent regulation, with the highes...

show abstract

Retinoic acid receptor expression vector inhibits differentiation of F9 embryonal carcinoma cells.

Cited by 72 publications

References 48 publications

Functional inhibition of retinoic acid response by dominant negative retinoic acid receptor mutants.

Functional inhibition of retinoic acid response by dominant negative retinoic acid receptor mutants.

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

Tissue-Dependent Expression of Heat Shock Factor 2 Isoforms with Distinct Transcriptional Activities

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