Functional analysis of the human estrogen receptor using a phenotypic transactivation assay in yeast

Pierrat, Benoı̂t; Heery, David M.; Lemoine, Yves; Losson, Régine

doi:10.1016/0378-1119(92)90277-v

Cited by 60 publications

(104 citation statements)

References 31 publications

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“…As in vertebrates (Chambon 1996 and references therein), the AF-1 and AF-2 of ER␣, RAR␣, and RXR␣ can activate transcription independently and synergistically in yeast White et al 1988;Pierrat et al 1992;Heery et al1993;and our unpublished results). Similarly, the transactivation potentials of ER␣ AF-1 and AF-2 are promoter-context-dependent in both animal and yeast cells (Tora et al 1989;Berry et al 1990;Metzger et al 1992).…”

supporting

confidence: 72%

supporting

confidence: 65%

“…Hollenberg (Vojtek et al 1993). The reporter strain PL3 (␣) [MAT␣,trp1::(ERE) 3 -URA3] was described elsewhere (Pierrat et al 1992). ⌬ada2, ⌬ada3, and ⌬gcn5 deletion strains were generated in the PL3 (␣) background according to the PCR-based disruption method described by Wach et al (1994).…”

Section: Methodsmentioning

confidence: 99%

“…The ADA3 gene was disrupted in the PL3 reporter strain, which contains a URA3 reporter gene controlled by three estrogen response elements (EREs, Fig. 4A; Pierrat et al 1992). Wild-type and ⌬ada3 PL3 strains were transformed with a high-copy-number plasmid YEp90 expressing [under the control of the phosphoglycerate kinase (PGK) promoter] either a chimeric receptor consisting of the DBD of ER␣ (ER.CAS, ER␣ amino acid residues 185-250) fused to the LBD/AF-2 of RXR␣ [RXR␣(DE)-ER.CAS; Heery et al 1993] or an amino-terminally truncated ER␣ derivative [ER␣(CDEF), previously called HEG19; Berry et al 1990] (Fig.…”

Section: Ada3 Acts As a Coactivator For The Af-2 Of Rxr␣ And Er␣ In Ymentioning

confidence: 99%

“…Receptor cDNAs used in this study correspond to human RAR␣1 and ER␣, mouse RXR␣ and chicken TR␣. Yeast expression plasmids YEp10 (TRP1), YEp90 (His3) and YCp90 (His3), have been described elsewhere (Pierrat et al 1992;Heery et al 1994). AAD (VP16) fusion proteins were expressed from the multicopy plasmid pASV3 (Leu2) (Le Douarin et al 1995b).…”

Section: Plasmidsmentioning

confidence: 99%

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The yeast Ada complex mediates the ligand-dependent activation function AF-2 of retinoid X and estrogen receptors

Baur¹,

Harbers²,

Um³

et al. 1998

Genes & Development

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Nuclear receptors (NRs) represent a large family of ligand-inducible transcriptional regulators that control complex developmental and homeostatic events in vertebrates by binding as homodimers or heterodimers to cognate DNA response elements present in target genes. NRs display a modular structure, with five to six distinct regions (denoted A-E/F; see Fig. 1A). The amino-terminal A/B region contains an autonomous activation function (AF-1), the highly conserved region C belongs to the DNA-binding domain (DBD), and the carboxy-terminal E region contains the ligand-binding domain (LBD), a dimerization surface and a ligand-dependent transcriptional activation function, AF-2 (for reviews and refs, see Mangelsdorf et al. 1995;Chambon 1996;Perlman and Evans 1997). The core of the AF-2 activating domain (AF-2 AD core; see Fig. 1A) has been characterized in the carboxy-terminal part of the E region and corresponds to the conserved amphipathic ␣ helix 12 of the LBD. Upon ligand binding, this helix is thought to fold back over the LBD to generate interacting surface(s) for the binding of transcriptional intermediary factors (TIFs, also denoted coactivators or mediators), whose effects ultimately result in the remodeling of the structure of the chromatin template and/or in the stimulation of initiation of RNA synthesis by the general transcription machinery (for reviews and references, see Chambon 1996;Wurtz et al. 1996;Glass et al. 1997).Although the yeast Saccharomyces cerevisiae does not possess endogenous NRs, it has been shown that a number of NRs, including the estrogen receptor (ER, now designated ER␣), the glucocorticoid receptor (GR), the retinoic acid (RA) receptors (RARs and RXRs), and the thyroid hormone receptor (TRs) can function as liganddependent transactivators in yeast (Metzger et al. , 1992Schena and Yamamoto 1988;Hall et al. 1993;Heery et al. 1993; and references therein). As in vertebrates (Chambon 1996 and references therein), the AF-1 and AF-2 of ER␣, RAR␣, and RXR␣ can activate transcription independently and synergistically in yeast White et al. 1988;Pierrat et al. 1992; Heery et al.1993; and our unpublished results

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supporting

confidence: 72%

supporting

confidence: 65%

Section: Methodsmentioning

confidence: 99%

Section: Ada3 Acts As a Coactivator For The Af-2 Of Rxr␣ And Er␣ In Ymentioning

confidence: 99%

Section: Plasmidsmentioning

confidence: 99%

See 3 more Smart Citations

The yeast Ada complex mediates the ligand-dependent activation function AF-2 of retinoid X and estrogen receptors

Baur¹,

Harbers²,

Um³

et al. 1998

Genes & Development

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Activation function 2 (AF-2) of retinoic acid receptor and 9-cis retinoic acid receptor: presence of a conserved autonomous constitutive activating domain and influence of the nature of the response element on AF-2 activity.

et al. 1994

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A motif essential for the transcriptional activation function 2 (AF‐2) present in the E region of retinoic acid receptor (RAR) alpha and 9‐cis retinoic acid receptor (RXR) alpha has been characterized as an amphipathic alpha‐helix whose main features are conserved between transcriptionally active members of the nuclear receptor superfamily. This conserved motif, which can activate autonomously in the absence of ligand in animal and yeast cells, can be swapped between nuclear receptors without affecting the ligand dependency for activation of transcription, thus indicating that a ligand‐dependent conformational change is necessary to reveal the AF‐2 activation potential within the E region of the nuclear receptor. Interestingly, we show that the precise nature of the direct repeat response element to which RAR/RXR heterodimers are bound can affect the activity of the AF‐2s of the heterodimeric partners, as well as the relative efficiency with which all‐trans and 9‐cis retinoic acids activate the RAR partner.

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The N-terminal part of TIF1, a putative mediator of the ligand-dependent activation function (AF-2) of nuclear receptors, is fused to B-raf in the oncogenic protein T18.

et al. 1995

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Nuclear receptors (NRs) bound to response elements mediate the effects of cognate ligands on gene expression. Their ligand‐dependent activation function, AF‐2, presumably acts on the basal transcription machinery through intermediary proteins/mediators. We have isolated a mouse nuclear protein, TIF1, which enhances RXR and RAR AF‐2 in yeast and interacts in a ligand‐dependent manner with several NRs in yeast and mammalian cells, as well as in vitro. Remarkably, these interactions require the amino acids constituting the AF‐2 activating domain conserved in all active NRs. Moreover, the oestrogen receptor (ER) AF‐2 antagonist hydroxytamoxifen cannot promote ER‐TIF1 interaction. We propose that TIF1, which contains several conserved domains found in transcriptional regulatory proteins, is a mediator of ligand‐dependent AF‐2. Interestingly, the TIF1 N‐terminal moiety is fused to B‐raf in the mouse oncoprotein T18.

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Functional analysis of the human estrogen receptor using a phenotypic transactivation assay in yeast

Cited by 60 publications

References 31 publications

The yeast Ada complex mediates the ligand-dependent activation function AF-2 of retinoid X and estrogen receptors

The yeast Ada complex mediates the ligand-dependent activation function AF-2 of retinoid X and estrogen receptors

Activation function 2 (AF-2) of retinoic acid receptor and 9-cis retinoic acid receptor: presence of a conserved autonomous constitutive activating domain and influence of the nature of the response element on AF-2 activity.

The N-terminal part of TIF1, a putative mediator of the ligand-dependent activation function (AF-2) of nuclear receptors, is fused to B-raf in the oncogenic protein T18.

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