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Mol Cell Biol, May 1998, p. 2949-2956, Vol. 18, No. 5
0270-7306/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.

Nuclear Receptor DAX-1 Recruits Nuclear Receptor Corepressor N-CoR to Steroidogenic Factor 1

Peter A. Crawford,1 Christoph Dorn,2 Yoel Sadovsky,2 and Jeffrey Milbrandt1,*

Departments of Pathology and Internal Medicine1 and Department of Obstetrics and Gynecology,2 Washington University School of Medicine, St. Louis, Missouri 63110

Received 1 December 1997/Returned for modification 26 January 1998/Accepted 13 February 1998

    ABSTRACT
Top
Abstract
Introduction
Materials & Methods
Results
Discussion
References

The orphan nuclear receptor steroidogenic factor 1 (SF-1) is a critical developmental regulator in the urogenital ridge, because mice targeted for disruption of the SF-1 gene lack adrenal glands and gonads. SF-1 was recently shown to interact with DAX-1, another orphan receptor whose tissue distribution overlaps that of SF-1. Naturally occurring loss-of-function mutations of the DAX-1 gene cause the human disorder X-linked adrenal hypoplasia congenita (AHC), which resembles the phenotype of SF-1-deficient mice. Paradoxically, however, DAX-1 represses the transcriptional activity of SF-1, and AHC mutants of DAX-1 lose repression function. To further investigate these findings, we characterized the interaction between SF-1 and DAX-1 and found that their interaction indeed occurs through a repressive domain within the carboxy terminus of SF-1. Furthermore, we demonstrate that DAX-1 recruits the nuclear receptor corepressor N-CoR to SF-1, whereas naturally occurring AHC mutations of DAX-1 permit the SF-1-DAX-1 interaction, but markedly diminish corepressor recruitment. Finally, the interaction between DAX-1 and N-CoR shares similarities with that of the nuclear receptor RevErb and N-CoR, because the related corepressor SMRT was not efficiently recruited by DAX-1. Therefore, DAX-1 can serve as an adapter molecule that recruits nuclear receptor corepressors to DNA-bound nuclear receptors like SF-1, thereby extending the range of corepressor action.

    INTRODUCTION
Top
Abstract
Introduction
Materials & Methods
Results
Discussion
References

Members of the nuclear receptor superfamily of transcription factors are critical players in a myriad of developmental, physiologic, and neoplastic processes (reviewed in references 7, 31, 39, 40, and 50). The modular structural motifs within nuclear receptors provide functional regions responsible for their activities. The domain structure typically consists of an amino-terminal ligand-independent activation domain (activation function 1 [AF-1], or domain A/B) that is usually not conserved among nuclear receptor subfamilies; a conserved DNA binding domain (DBD) (domain C), consisting of two zinc-binding modules; an intervening hinge region (domain D); and a carboxy-terminal ligand binding domain (domain E). Domain E participates in receptor hetero- or homodimerization (8, 46), as well as in transcriptional repression and ligand-induced transcriptional activation (4, 12, 16, 22, 37). In fact, domains D and E serve as the interface for a multitude of cooperating proteins whose purpose is to transduce activating or repressive transcriptional signals (reviewed in reference 23). Corepressor molecules N-CoR (nuclear receptor corepressor) and SMRT (silencing mediator of retinoid and thyroid receptors) usually dissociate from receptor when ligand is bound (12, 22, 33). N-CoR and SMRT repress transcription by recruiting a complex of proteins, including mSin3A and mRPD3 (HDAC1), which in turn deacetylate histones (1, 20, 45). Orphan nuclear receptors, which have no known high-affinity endogenous ligands, also interact with corepressor molecules (19, 66, 67), but the mechanisms which regulate those interactions are not known. While it is possible that the associations are constitutive, it is likely that unknown ligands and/or other interactive proteins influence these interactions.

The nuclear receptor steroidogenic factor 1 (SF-1) is an orphan receptor, and it is able to bind DNA with high affinity as well as activate transcription as a monomer (63). While 25-hydroxycholesterol has been shown to potentiate SF-1 activity (34), no high-affinity-specific ligand has been isolated. The SF-1 gene is expressed constitutively in specific endocrine tissues: the hypothalamus, anterior pituitary, adrenal gland, gonads, and placenta (26, 27, 29, 51). The importance of SF-1 is underscored by its ability to regulate the expression of many genes characteristic of these tissues. For instance, experiments with cultured cell lines have shown that each steroidogenic enzyme gene and the steroidogenic acute regulatory protein are regulatable by SF-1 (3, 11, 13, 21, 25, 27, 36, 41-43, 49, 58, 64, 70). Moreover, SF-1 is a regulator of the genes encoding Müllerian inhibitory substance in vitro and in vivo (17, 55), the alpha -subunit of the gonadotropins (6), the beta -subunit of luteinizing hormone (18, 32, 35), the adrenocorticotropin receptor (10), the prolactin receptor (24), and oxytocin (62). Furthermore, SF-1 is of particular interest biologically because of its integral role in mammalian development: it serves as a critical regulator of the development of the adrenal glands, gonads, and ventromedial hypothalamus; additionally, it is a dominant coordinator of the steroidogenic cell phenotype (14, 26, 38, 51, 57). Much remains unknown about SF-1, however, such as target genes in the developing urogenital ridge, as well as the molecules which modify its activity at specific spatial and temporal locations in development. Such molecules could contribute to the diversity of functions exhibited by this transcription factor.

DAX-1 is an atypical orphan receptor, harboring a conserved domain E, but instead of domains A to D, it possesses an amino-terminal domain that harbors repetitive regions rich in glycine and alanine residues. The gene encoding DAX-1 was positionally cloned as the gene mutated in X-linked adrenal hypoplasia congenital (AHC), a birth defect characterized by adrenal insufficiency and hypogonadotropic hypogonadism at puberty (44, 68). The expression of DAX-1 (for dose-sensitive sex reversal-AHC critical region on the X chromosome, gene 1) in the mouse overlaps that of SF-1, namely, the hypothalamus, pituitary, adrenal gland, and gonads (28, 59). Furthermore, the similarity of the phenotype of AHC patients to that of mice who lack SF-1 suggests that these receptors cooperate in development. Paradoxically, DAX-1 is an inhibitor of steroidogenesis, because it suppresses transcriptional activation by SF-1 through direct physical interaction (30), and DAX-1 serves as a DNA-bound repressor in some promoter contexts (69). Because the developmental and physiologic roles of DAX-1 and SF-1 appear to be interrelated, it is important to determine what mechanism DAX-1 employs to repress transcription.

To dissect the transcriptional activity of the SF-1-DAX-1 complex, we have localized the domain within SF-1 through which DAX-1 interacts. Furthermore, we have found that DAX-1 recruits the nuclear receptor corepressor N-CoR to SF-1, which otherwise do not interact. However, AHC mutations of DAX-1 do not readily recruit N-CoR to SF-1, thus decreasing their ability to repress SF-1 activity. These findings extend the range of corepressor action by demonstrating that a nuclear receptor normally refractory to repression by N-CoR (i.e., SF-1) is susceptible in the presence of DAX-1. DAX-1 is therefore a potentially important variable in corepressor signaling.

    MATERIALS AND METHODS
Top
Abstract
Introduction
Materials & Methods
Results
Discussion
References

Plasmids. All cloning that employed PCR was performed with the high-fidelity KlenTaq polymerase enzyme (5). DAX-1 was cloned by reverse transcription-PCR from mouse adrenal gland total RNA into pBSKS (Stratagene). The template was sequenced, compared to the mouse DAX-1 sequence (2, 59), and then used for cloning into subsequent vectors. Murine SF-1 cDNA was obtained and used as previously described (14, 15, 65). Murine RARalpha cDNA was obtained from S. Adler (Washington University, St. Louis, Mo.). pCMX-N-CoR (amino acids 1 to 2453) was obtained from A. Hörlein (DKFC, Heidelberg, Germany); pCMX-N-CoR (amino acids 1510 to 2453) was obtained from M. Lazar (University of Pennsylvania). pCMX-SMRT was obtained from J. D. Chen (University of Massachusetts). GAL4 DBD (amino acids 1 to 147) fusions were made in the pM vector series (52). Fragments of the SF-1 ligand binding domain (LBD) (amino acids 120 to 462, 120 to 451, 120 to 447, 120 to 437, 120 to 380, 120 to 277, 220 to 462, 226 to 462, 230 to 462, and 245 to 462), DAX-1 LBD (amino acids 256 to 472 and 256 to 369), or RARalpha (amino acids 162 to 462) were generated by PCR and cloned in frame into the appropriate pM vector. Full-length or mutant cDNAs were expressed from the pCMVneo vector (9). For VP16 fusions, full-length DAX-1 (amino acids 1 to 472), partial DAX-1 (amino acids 1 to 369), or N-CoR (amino acids 1550 to 2453) was amplified by PCR and cloned downstream of and in frame with VP16 (residues 411 to 487), which harbors a synthetic Kozak initiator methionine within pCMVneo. The SMRT gene (encoding amino acids 565 to 1495) was amplified with a primer providing a synthetic initiator methionine and cloned upstream of VP16 in pCMVneo. Point mutagenesis of DAX-1 was performed by inverse PCR, after which template DNA was removed by digestion with DpnI (New England Biolabs, Inc.). Mutants R267P and F449D of DAX-1 correspond to the human DAX-1 numbering; for the mouse sequence used in these studies, the mutations are actually R269P and F451D.

Transfected SF-1 activity was measured from the SF-1 luciferase reporter (SF-1/Luc), which harbors two SF-1 binding sites (5'-TCA AGGTCA) spaced by five nucleotides upstream of the prolactin minimal promoter and the luciferase reporter gene (a derivative of pPrl36 [15]). For GAL4 fusions, activity was measured from the plasmid Delta GKI (obtained from P. Webb, University of California, San Francisco), which harbors five GAL4 binding sites upstream of the thymidine kinase minimal promoter and luciferase (15).

Transfections. CV-1 monkey kidney cells were grown in Dulbecco's modified Eagle medium, 10% fetal bovine serum, and antibiotics at 5% CO2. JEG-3 human choriocarcinoma cells were maintained in minimal essential medium with Earle's salts, 10% fetal bovine serum, and antibiotics at 5% CO2. Transfections were performed in 12-well plates (cells plated at 5 × 104/well 24 h prior to addition of DNA) by calcium phosphate precipitation in CV-1 media, and JEG-3 cells were placed in 10% CO2 for 3 h prior to the addition of DNA. Cells were harvested 48 h after the addition of DNA (1 µg of total DNA per well), and lysates were analyzed for luciferase activity as previously described (15). For each experiment, transfections were performed at least five times in duplicate. All data sets presented yielded <= 10% variation between duplicates. In each case, the results presented are of one representative experiment. All luciferase activities were normalized to beta -galactosidase activity, derived from a cotransfected pRSV-beta -gal plasmid. For two-hybrid experiments performed with GAL4 DBD fusions, all fold activities were corrected for background reporter activity derived from expression plasmids cotransfected with GAL4 DBD alone (pM2 vector). For experiments performed with native SF-1, fold activation levels were corrected for background activity derived from expression plasmids cotransfected with nonrecombinant pCMVneo.

Physical interaction assay. All proteins were translated in vitro with the TNT in vitro transcription-translation kit (Promega), in the presence of [35S]Met (DuPont/NEN). Partial N-CoR was translated from pCMX-N-CoR (amino acids 1510 to 2453). GAL4 DBD fusions of DAX-1 were translated from pCITE 3 (Novagen, Inc.). Protein A-agarose beads (Gibco BRL, Inc.) were incubated overnight at 4°C with mouse anti-GAL4 DBD antibody (sc-510 [Santa Cruz]) at a final concentration of 15 µg/ml. After being washed in immunoprecipitation (IP) buffer (20 mM HEPES-KOH [pH 7.9], 100 mM KCl, 1 mM EDTA, 5% glycerol, 0.5% Nonidet P-40, 1 mM dithiothreitol, 1 mM phenylmethylsulfonyl fluoride), beads were incubated for 20 min with 500 µg of bovine serum albumin per ml at 4°C. Three microliters of each in vitro-translated protein was incubated at 4°C in 200 µl of IP buffer. After 20 min, 30 µl of blocked antibody-protein A-agarose conjugate was added to each reaction mixture, which was allowed to incubate at 4°C rocking for 2 h. Beads were washed four times with IP buffer and incubated for 5 min with 30 µl of 2× sodium dodecylsulfate-polyacrylamide gel electrophoresis (SDS-PAGE) loading buffer at 70°C, and eluates were electrophoresed. After signal enhancement, gels were dried down and exposed to film.

Western blotting. COS-1 cells were seeded in six-well plates in CV-1 media at 1.5 × 105 cells/well. Twenty-four hours later, cells were transfected with expression vectors encoding GAL4 DBD-DAX-1 LBD fusion proteins. Forty-eight hours after the addition of DNA, cells were lysed in radioimmunoprecipitation assay buffer (1× phosphate-buffered saline, 0.1% Nonidet P-40, 0.5% sodium deoxycholate, 0.1% SDS, 1 mM EDTA, 1 mM phenylmethylsulfonyl fluoride). Ten micrograms of total cell lysate was subjected to SDS-PAGE, transferred to nitrocellulose, and blotted with the anti-GAL4 DBD antibody used above, by using a horseradish peroxidase-conjugated goat anti-mouse secondary antibody. The signal was developed by enhanced chemiluminescence (Amersham) and exposed to film for 5 min.

    RESULTS
Top
Abstract
Introduction
Materials & Methods
Results
Discussion
References

Repression of SF-1 by DAX-1 requires an SF-1-repressive domain between residues 437 and 447. To assess the ability of DAX-1 to repress SF-1 transcriptional activation, we cotransfected JEG-3 human choriocarcinoma cells with wild-type SF-1 and increasing concentrations of mouse DAX-1 (DAX-1 does not bind to an SF-1 response element [data not shown and reference 30]). In addition, we employed two mutated versions of DAX-1 which correspond to naturally occurring human mutations that cause AHC (R267P and del 369 [44]). With an SF-1-responsive reporter, wild-type DAX-1 was a potent repressor of SF-1 activity, whereas the naturally occurring DAX-1 mutants were much less potent SF-1 repressors (Fig. 1A).


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  		FIG. 1.   Repression of SF-1 transactivation by DAX-1 requires the most carboxy-terminal repressive domain within SF-1. (A) JEG-3 cells were cotransfected with full-length SF-1 (50 ng) and increasing amounts of the wild type, mutant R267P, or mutant del 369 DAX-1 (0, 5, 15, or 100 ng), along with 250 ng of the SF-1/Luc reporter, which harbors two SF-1 response elements upstream of the prolactin minimal promoter TATA box. (B) JEG-3 cells were cotransfected with empty expression vector (control), full-length SF-1 (amino acids 1 to 462), SF-1 (amino acids 1 to 451), or SF-1 (amino acids 1 to 277) (30 ng) in the presence or absence of exogenous wild-type DAX-1 (10 ng) and 250 ng of SF-1/Luc reporter. Luciferase activities were measured and standardized as described in Materials and Methods.

Because SF-1 does not possess an AF-1 domain (domain A/B), it relies entirely on the carboxy terminus (domains D and E) to modulate transcription. Having characterized two regions responsible for transcriptional activation (15), we used progressive truncations to unveil regions that mediate trans-repression. These experiments illustrated that the region including SF-1 amino acids 277 to 451 possesses repressive activity (data not shown). To determine if repression of SF-1 by DAX-1 requires this portion of the SF-1 LBD, we transfected full-length SF-1 (amino acids 1 to 462), the AF-2-AH deletion (amino acids 1 to 451), or the truncation (amino acids 1 to 277) into JEG-3 cells and determined their abilities to be repressed by DAX-1 (wild type). Full-length SF-1 (amino acids 1 to 462) or SF-1 (amino acids 1 to 451) is repressed by DAX-1, but SF-1 (amino acids 1 to 277) is not (Fig. 1B). DAX-1 could similarly repress fusions of these SF-1 truncations to the VP16 activation domain (data not shown), a heterologous domain that fully restores transcriptional activation of SF-1 derivatives which lack the carboxy-terminal AF-2-AH (14). Because DAX-1 is unable to repress SF-1 which lacks this carboxy-terminal domain (between residues 277 and 451), DAX-1 most likely mediates repression of SF-1 through this region.

To determine if SF-1 amino acids 277 to 451 are in fact required for a physical interaction between SF-1 and DAX-1, we performed two-hybrid experiments with CV-1 cells by cotransfecting GAL4-SF-1 carboxy-terminal fusions along with the entire DAX-1 cDNA fused to the VP16 activation domain. In addition, we utilized fusions of VP16 and the two DAX-1 AHC mutants used above to confirm that they were in fact capable of interacting with SF-1 (30). As measured by activity from the GAL4 reporter, fusions of GAL4 DBD to SF-1 amino acids 120 to 462, 120 to 451, and 120 to 447 were all capable of strong interactions with all three versions of DAX-1 (only VP16-DAX-1 [wild type] is shown for the GAL4-SF-1 [amino acids 120 to 462] construct) (Fig. 2A). The magnitude of the SF-1-DAX-1 interaction was diminished with the AHC-inducing mutants of DAX-1, but could be partially overcome by increasing the amount of VP16-DAX-1 mutant (only the higher concentration of VP16-DAX-1 mutants is shown). In addition, SF-1 LBD fusions which lack AF-2-AH exhibited enhanced interaction with DAX-1, which is a feature that other nuclear receptor-corepressor interactions have manifested (19). The fusion protein containing SF-1 amino acids 120 to 442 interacted weakly with all three VP16-DAX-1 partners, and those containing SF-1 amino acids 120 to 437 or 120 to 380 did not interact at all with VP16-DAX-1 (wild type or mutant). Thus, the SF-1 region circumscribed by amino acids 437 to 447 (termed the R domain [YLYHKHLGNEM]) is required for interaction with, and therefore repression by, DAX-1. Furthermore, while strong interactions occur between SF-1 and DAX-1 AHC mutants, a component of the diminished repression of SF-1 by AHC mutants of DAX-1 may also be attributable to attenuated interaction with SF-1 (see Discussion).


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  		FIG. 2.   Interaction between SF-1 and DAX-1 requires two domains within SF-1. CV-1 cells were cotransfected with fusions of GAL4 DBD and SF-1 carboxy terminus (20 ng), fusions of VP16 and DAX-1 (wild type [amino acids 1 to 472]), DAX-1 (R267P), or DAX-1 (del 369) (50 or 200 ng), and 250 ng of GAL4 reporter. (A) Fusions of GAL4 DBD with SF-1 amino acids 120 to 462, 120 to 451, 120 to 447, 120 to 442, 120 to 437, or 120 to 380 were tested with a two-hybrid assay against all three forms of VP16-DAX-1. Only activity from VP16-DAX-1 (wild type) is shown for GAL4-SF-1 (amino acids 120 to 462). (B) Fusions of GAL4 DBD with SF-1 amino acids 120 to 462, 220 to 462, 226 to 462, 230 to 462 or 245 to 462 were tested in a two-hybrid assay against VP16-DAX-1 (wild type). Luciferase activities were measured and standardized as described in Materials and Methods. Fold activities were calculated by determining the degree of enhancement by VP16-DAX-1 of the activity of each GAL4-SF-1 construct compared to the degree of enhancement observed with GAL4 DBD (see Materials and Methods). (C) Schematic diagram of SF-1, demonstrating the DBD, T/A box, PID, R domain, and AF-2-AH. See text for details.

Previously, we determined that two distinct domains within the SF-1 carboxy terminus were required for the interaction of SF-1 with and potentiation by the coactivator SRC-1 (15). Therefore, we questioned whether or not the interaction of SF-1 with DAX-1 involved an analogous arrangement of multiple domains. To determine if additional regions from the hinge or LBD of SF-1 were required for interaction with DAX-1 in concert with the R domain, we made progressive amino-terminal truncations from the SF-1 hinge or LBD, maintaining the natural stop codon at the carboxy terminus (after residue 462). Using the two-hybrid assay with CV-1 cells, we demonstrated that fusions of GAL4 DBD to SF-1 residues 120 to 462, 220 to 462, and 226 to 462 were able to interact with VP16-DAX-1 (wild type), as well as with the VP16 fusion to mutants of DAX-1 (Fig. 2B and data not shown). However, fusions of the GAL4 DBD to SF-1 residues 230 to 462 or 245 to 462 were unable to interact with VP16-DAX-1 (Fig. 2B). Therefore, in addition to the R domain of SF-1, residues 226 to 230 (ELILQ) are required for the interaction between SF-1 and DAX-1. Interestingly, we had previously shown that residues 187 to 245 of SF-1 are required for the interaction between SF-1 and the coactivator SRC-1 (15). Further delimitation of this domain has shown that the SF-1-SRC-1 interaction in fact requires the same residues (226 to 230) as those required for the SF-1-DAX-1 interaction (49a). Thus, the region between amino acids 226 and 230 (proximal interactive domain [PID]) mediates interactions with both SRC-1 and DAX-1 (see Discussion). A schematic of the SF-1 functional domains incorporates these regions (Fig. 2C).

DAX-1, but not AHC mutants, recruits N-CoR to SF-1. DAX-1 represses SF-1 by interacting with specific domains within SF-1, but the mechanism of repression by DAX-1 is unclear. While the ability of AHC mutants to interact with SF-1 is partially abrogated, the repression of SF-1 by these mutants is more severely affected than this diminished interaction would produce, suggesting that AHC mutations may also attenuate the actual repression function of DAX-1. All AHC mutations discovered to date mutate or eliminate the LBD of DAX-1 (44, 68), indicating that the DAX-1 LBD may transduce a repression function. To determine if DAX-1 LBD acts as a repressor of transcription when bound directly to DNA, we fused DAX-1 LBD (amino acids 256 to 472) to GAL4 DBD (amino acids 1 to 147) and compared its transcriptional activity to that of GAL4 DBD alone on a GAL4 reporter in CV-1 cells. Additionally, we transfected fusions of GAL4 DBD and DAX-1 LBD that correspond to naturally occurring AHC mutations (amino acids 256 to 472 [R267P] and 256 to 369). As shown in Fig. 3, GAL4-DAX-1 (amino acids 256 to 472) represses basal transcriptional activity, but fusions of DAX-1 which harbor AHC LBD mutations do not readily repress. Thus, the DAX-1 LBD harbors a repression function, and mutations of DAX-1 that cause AHC disturb this repressive ability. A Western blot demonstrates the equivalent stability of these transfected GAL4-DAX-1 fusions (see Fig. 6E). A similar blot demonstrated the equivalent stability of the GAL4-SF-1 fusions described above (data not shown).


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  		FIG. 3.   DAX-1 acts as a repressor of transcription when tethered to DNA. Expressors driving GAL4 DBD alone or fusions of GAL4 DBD and DAX-1 (amino acids 256 to 472, 256 to 472 [R267P], and 256 to 369) (20 ng) were transfected with 250 ng of GAL4 reporter as described in Materials and Methods. Fold repression relative to the activity that observed with GAL4 DBD is given in parentheses. Luciferase activities were determined and standardized as described above.

The mechanism of transcriptional repression by DAX-1 is unknown. Several nuclear receptors repress transcription through a mechanism that involves the recruitment of the nuclear receptor corepressors N-CoR and SMRT (see the introduction). To determine if DAX-1 shares this function, we performed two-hybrid experiments with CV-1 cells by using GAL4-DAX-1 LBD (amino acids 256 to 472) and VP16-N-CoR (amino acids 1550 to 2453) or VP16-SMRT (amino acids 565 to 1495) fusion proteins. These fragments of N-CoR or SMRT harbor regions previously shown to be required for interaction with nuclear receptors (54, 56). For a positive control, we utilized a GAL4-RARalpha fusion, which demonstrates a >20-fold interaction with VP16-N-CoR or VP16-SMRT in the absence of ligand (all-trans-retinoic acid [atRA]), but no interaction in the presence of ligand. While wild-type DAX-1 LBD interacts with N-CoR, two mutants of DAX-1 with mutations which correspond to the AHC-inducing mutations (amino acids 256 to 472 [R267P] and 256 to 369) do not (Fig. 4A). Interestingly, DAX-1 (wild type) and SMRT do not readily interact, which is analogous to the relationship between RevErb (as well as RORalpha ) and these corepressors (references 19 and 67 and as described below). Furthermore, the interaction between DAX-1 and N-CoR is weaker than that between unliganded RARalpha and N-CoR, requiring a higher concentration of added VP16-N-CoR to obtain an interaction, which could indicate that the complex between DAX-1 and N-CoR is weaker and may require additional stabilizing proteins (see Discussion).


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  		FIG. 4.   DAX-1 LBD interacts with N-CoR. (A) CV-1 cells were cotransfected with fusions of GAL4 DBD and RARalpha (amino acids 162 to 462) or DAX-1 LBD (amino acids 256 to 472, 256 to 472 [R267P], or 256 to 369 [del 369]) (20 ng), fusions of VP16 and N-CoR (1550 to 2453) or SMRT (565 to 1495) (20 or 800 ng), and 200 ng of GAL4 reporter. Luciferase activities were measured and standardized as described in Materials and Methods. Fold activities were calculated by determining the degree of enhancement by VP16-N-CoR or VP16-SMRT of the activity of each GAL4-DAX-1 (or GAL4-RARalpha ) construct, compared to the degree of enhancement observed with GAL4 DBD (see Materials and Methods). (B) In vitro interaction between N-CoR and nuclear receptors. The experiment was performed as described in Materials and Methods. Precipitation of N-CoR (amino acids 1510 to 2453) by RARalpha (as well as 1/10 loaded input N-CoR) was exposed to film for 15 min. Precipitation of N-CoR by the DAX-1 wild type (amino acids 256 to 472) or mutant (amino acids 256 to 369) was exposed to film for 2 h. Where indicated, 10-7 M atRA was added to transfection media or to the protein incubation. M, molecular mass.

The two-hybrid interaction in mammalian cells between DAX-1 and N-CoR was confirmed with a physical interaction assay. An in vitro-translated N-CoR fragment (amino acids 1510 to 2453) was tested for binding to GAL4 DBD fusions of DAX-1 in a coimmunoprecipitation assay with an anti-GAL4 DBD antibody. As demonstrated with two-hybrid interaction, the GAL4-DAX-1 wild type (amino acids 256 to 472) precipitates N-CoR more readily than the GAL4-DAX-1 mutant (amino acids 256 to 369), but less readily than RARalpha (Fig. 4B). The in vitro relationship between wild-type and mutated DAX-1 is similar to that between unliganded and liganded RARalpha (incubated with 10-7 M atRA). The interaction between N-CoR and RARalpha in vitro is more robust than that between N-CoR and DAX-1, because the N-CoR and DAX-1 lanes were exposed to film for a longer period.

Thus far, we have demonstrated that GAL4 fusions of DAX-1 interact with the corepressor N-CoR. To determine if wild-type DAX-1 is able to recruit N-CoR to SF-1, we performed a modified two-hybrid experiment between SF-1 and N-CoR. We tested several of the previously employed GAL4 DBD fusions of SF-1 (amino acids 120 to 462, 120 to 447, 120 to 437, and 230 to 462) for the ability to interact with fusions of VP16 and N-CoR (amino acids 1550 to 2453) in the presence or absence of exogenous full-length wild-type or mutated DAX-1. The SF-1 LBD (amino acids 120 to 462) was unable to significantly interact with N-CoR, unless wild-type DAX-1 was added, which permitted an ~20-fold increase (Fig. 5). On the other hand, AHC mutants of DAX-1 were not able to potentiate the SF-1-N-CoR interaction, even at high concentrations of added mutant DAX-1. Fusion of SF-1 amino acids 120 to 447, a sequence which lacks AF-2-AH, permitted very weak interaction with N-CoR in the absence of added DAX-1, but this interaction was enhanced when DAX-1 was added. Finally, the DAX-1-enhanced SF-1-N-CoR interaction explicitly requires the same domains of SF-1 which are required for an SF-1-DAX-1 interaction, because removal of the R domain (achieved with the amino acids 120-to-437 construct) or the PID (achieved with the amino acid 230-to-462 construct) abrogates interaction between SF-1 and N-CoR in the presence or absence of added DAX-1 (Fig. 5). In a control experiment, added full-length N-CoR did not modify the two-hybrid interaction between GAL4-SF-1 and VP16-DAX-1 (data not shown). It is important to note that the potentiation of the SF-1-N-CoR interaction requires low levels of added DAX-1 and less VP16-N-CoR than the direct DAX-1-N-CoR two-hybrid assay.


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  		FIG. 5.   DAX-1 potentiates two-hybrid interaction between SF-1 and N-CoR. CV-1 cells were cotransfected with GAL4 DBD fusions of the SF-1 carboxy terminus (amino acids 120 to 462, 120 to 447, 120 to 437, or 230 to 462) (20 ng), VP16-N-CoR (20 ng [Fig. 5]); increasing amounts of wild-type, mutant R267P, or del 369 DAX-1 (0, 5, 10, 15, or 100 ng); and GAL4 reporter (250 ng). Luciferase activities were measured and standardized as described in Materials and Methods. Fold activities were calculated by determining the degree of enhancement by VP16-N-CoR of the activity of each GAL4-SF-1 construct in the presence or absence of DAX-1 constructs compared to the degree of enhancement observed with GAL4 DBD (see Materials and Methods).

Conservation of interactive motifs confirms DAX-1-N-CoR interaction. The specific interaction of DAX-1 with N-CoR but not SMRT also occurs with nuclear receptors RevErb and RORalpha , which were shown to interact with both N-CoR and SMRT in glutathione S-transferase precipitation assays but with only N-CoR in DNA-dependent binding assays (19, 67). To determine if RevErb and DAX-1 employ a similar mechanism of corepressor recruitment, we examined the two N-CoR-interactive domains delimited within RevErb LBD (amino acids 407 to 418 and 602 to 614 [66, 67]) and compared them to regions within DAX-1 that might be involved with a DAX-1-N-CoR interaction (Fig. 6A). The amino-terminal interactive domain of RevErb LBD shares homology with two residues within the DAX-1 LBD that are mutated in some AHC kindreds (44), R267 (conserved within RevErb as K412) and V269 (conserved with RevErb as V414). Furthermore, the R267P DAX-1 mutant is incapable of efficient SF-1 repression as well as N-CoR recruitment (Fig. 2A and 6). The carboxy-terminal N-CoR interactive domain of RevErb shares three conserved residues within the carboxy terminus of DAX-1, which is often deleted in AHC kindreds. Strikingly, mutation of a conserved Phe residue in this domain of DAX-1 (F449D) abrogates the repression function of GAL4-DAX-1 (amino acids 256 to 472) and completely abolishes the two-hybrid interaction between DAX-1 and N-CoR (Fig. 6B and data not shown). Moreover, DAX-1's ability to potentiate the two-hybrid interaction between GAL4-SF-1 (amino acids 120 to 462) and VP16-N-CoR is diminished when DAX-1 (F449D) is added in lieu of wild-type DAX-1 (Fig. 6C). Finally, DAX-1 (F449D) is a less potent repressor of SF-1-mediated transcriptional activation in JEG-3 cells than wild-type DAX-1, behaving similarly to the AHC DAX-1 mutants (Fig. 6D). All three assays confirm the required integrity of this carboxy-terminal domain for repression by DAX-1. Therefore, RevErb and DAX-1 probably employ similar motifs to interact with N-CoR, and their conservation supports the specificity of corepressor recruitment exhibited by each of them. To confirm that each mutation of DAX-1 does not diminish the stability of the resulting proteins, we performed Western blotting of transfected fusions of GAL4 DBD and wild-type DAX-1, as well as mutants R267P, del 369, and F449D. These of DAX-1 mutants do not diminish protein stability (Fig. 6E). Therefore, the inability of DAX-1 mutants to repress SF-1 and recruit N-CoR reflects an impairment in the execution of normal DAX-1 function.


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  		FIG. 6.   Conservation of N-CoR-interactive domains within DAX-1 and RevErb. (A) Two of the three previously delimited domains within RevErb required for interaction with N-CoR (66, 67) share similarities with regions of DAX-1 which are mutated in AHC and which are also required for interaction with N-CoR. Arrows demarcate residues analyzed in this study: R267 of DAX-1 is mutated in some AHC kindreds (R267P), and F449 is one of three shared residues in the carboxy-terminal-interactive domain of RevErb (B, C, and D). (B) Mutation F449D prevents the two-hybrid interaction between DAX-1 and N-CoR. A two-hybrid experiment was performed and results were analyzed as described in the legend to Fig. 5A (500 ng of VP16-N-CoR used in this experiment). (C) Mutation F449D in the context of full-length DAX-1 abrogates the potentiation of the two-hybrid SF-1-N-CoR interaction. Ten nanograms of DAX-1 (wild type [WT]) or F449D was transfected with 20 ng of VP16-N-CoR, 20 ng of GAL4-SF-1 (120 to 462), and 200 ng of GAL4 reporter. (D) Mutant F449D of DAX-1 is not a potent repressor of SF-1 transactivation. Fifty nanograms of SF-1 expressor was cotransfected with 20 ng of DAX-1 (wild-type) or DAX-1 (F449D) mutant, along with 250 ng of SF-1/Luc reporter. The experiment was analyzed as described in the legend to Fig. 2A. (E) Western blotting (performed as described in Materials and Methods with anti-GAL4 DBD antibody) demonstrates expression levels of GAL4 DBD-DAX-1 mutant fusions. Lanes: 1, GAL4-DAX-1 (amino acids 256 to 472); 2, GAL4-DAX-1 (amino acids 256 to 472 [R267P]); 3, GAL4-DAX-1 (amino acids 256 to 369); 4, GAL4-DAX-1 (amino acids 256 to 472 [F449D]). MW, molecular mass.

    DISCUSSION
Top
Abstract
Introduction
Materials & Methods
Results
Discussion
References

DAX-1 is a structurally unique nuclear receptor whose gene is mutated in the human disorder AHC. The amino-terminal region of DAX-1 does not harbor a classical DBD, but instead possesses a structurally uncharacterized region that binds DNA (69). The conserved LBD (domain E) is usually deleted or mutated in AHC-inducing DAX-1 alleles (44, 68). Because nuclear receptor domains D and E provide surfaces for interactions with a variety of proteins, including coactivators, corepressors, and molecules with unidentified function (23), domain E of DAX-1 may be an important target for molecules which influence fundamental developmental and physiologic processes. The results presented herein demonstrate that DAX-1 is able to repress transcriptional activation by SF-1 and is also able to interact with the nuclear receptor corepressor N-CoR. Collectively, less-repressive AHC mutations in DAX-1 leave intact the ability to bind DNA (69) as well as interact with SF-1 (shown herein and within reference 30) but disrupt the ability of DAX-1 to interact with N-CoR. Therefore, a primary responsibility of DAX-1 in the developing adrenal gland may be to recruit N-CoR, or a related corepressor, to appropriate target promoter contexts. Absence of corepressor recruitment may in turn result in aberrant maturation of the adrenocortical phenotype. While seemingly paradoxical, the requirement for transcriptional repression in development has precedence, particularly in Drosophila melanogaster with proteins such as Krüppel (47, 53). While corepressors may be presumed to play critical roles in cycling physiologic cascades, as well as in genetic mechanisms that protect against neoplastic events, loss-of-function studies of mice targeted for deletion of N-CoR and/or SMRT could also unveil developmental roles in the genesis of many tissue types.

The interaction among SF-1, DAX-1, and N-CoR requires a number of domains within each molecule. To interact with DAX-1, SF-1 requires at least two domains: a carboxy-terminal repressive domain (R domain [amino acids 437 to 447]) and a PID that was previously shown to be required for transcriptional activation by SF-1 through interaction with the SRC-1 family of coactivators (reference 15 and data not shown). Thus, the PID (which lies within the amino terminus of domain E) is required for SF-1 to interact with both the activator and repressor molecules. It is not known whether the interactions between SF-1 and SRC-1 or SF-1 and DAX-1 are mutually exclusive. The PID may fold as part of a tertiary structure that allows other motifs within SF-1 to fold correctly and serve as interfaces with other regulatory molecules. The role of intramolecular contacts in the generation of higher-order structures within nuclear receptor LBDs has strong precedence among the retinoid and thyroid receptors (8, 48, 61). To interact with SF-1, DAX-1 uses its DBD (30), while the repression function of DAX-1 is mediated by at least two regions of the LBD, both of which are required to recruit N-CoR to DNA. Interestingly, both the R domain of SF-1 and the carboxy-terminal N-CoR interaction domain within DAX-1 (Fig. 6A) lie within helix 11 of the LBD, according to alignments and structural analyses of RXRalpha and RARgamma (8, 48). The sequences of these regions within SF-1 and DAX-1 are not identical, however, which is consistent with the distinct function of these two domains. A schematic model for the interaction among SF-1, DAX-1, and N-CoR is shown in Fig. 7. The attenuated ability of DAX-1 AHC mutants to repress SF-1 clearly arises because of a loss of repressive function by the DAX-1 LBD (Fig. 3), which is in turn caused by an inability to recruit N-CoR (Fig. 4 and 5). However, our results also suggest that a component of the weakened repression of SF-1 by AHC mutants of DAX-1 may be due to a diminished ability to interact with SF-1 (Fig. 2B). Indeed, the interaction among SF-1, DAX-1, and N-CoR may be interdependent and could involve other nuclear proteins that govern the stability of the putative complex.


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  		FIG. 7.   Schematic model of mechanism of DAX-1 repression of SF-1. SF-1, bound to its response element on DNA, recruits DAX-1, requiring the PID and the R domain within the carboxy terminus of SF-1. To interact with SF-1, DAX-1 employs its structurally degenerate DBD (30). DAX-1 in turn recruits N-CoR to DNA, requiring at least two conserved regions within DAX-1 LBD, either of which is mutated or eliminated in different AHC kindreds. Other molecules may stabilize or modify the interaction among the three proteins, depending on the cell or promoter context.

Despite the clear ability of wild-type DAX-1 to interact with N-CoR, the magnitude of interaction was nevertheless lower than that of unliganded RARalpha with N-CoR in two-hybrid as well as physical interaction assays. There are several potential explanations for this finding. First, the interaction may be stabilized in cells by unknown proteins involved in DAX-1 or N-CoR signaling. Second, posttranslational modification, such as phosphorylation, of DAX-1 or N-CoR may modify the interaction. Third, unknown N-CoR and SMRT family members could serve as higher-affinity targets for DAX-1. Irrespective of the precise reason, influences over receptor-corepressor interaction most likely extend beyond the known roles of ligand-induced conformational changes, as well as polarity dependence of DNA response elements (12, 22, 33). For those receptors which may have no high-affinity endogenous ligand, other factors are likely to hold influence. These factors could plausibly play roles in specific cell types under particular physiologic conditions, allowing for greater DAX-1-N-CoR interaction in vivo.

Recent studies have shown that DAX-1 is a transcriptional repressor and inhibitor of steroidogenesis (30, 68, 69). Nevertheless, the AHC phenotype (which is caused by loss-of-function mutations of DAX-1) suggests that the role of DAX-1 in the developing and mature urogenital ridge, as well as the hypothalamus and pituitary, most likely also influences processes outside the regulation of steroidogenesis. Because DAX-1 is able to repress transcription when bound to hairpin structures of DNA (69), or when indirectly bound through another transcription factor, like SF-1, its scope of action is likely to be fairly large. Because DAX-1 has been shown to be an inhibitor of retinoid transcriptional activation as well (68), it will be interesting to determine if the DBD of DAX-1 permits interaction with other receptors, extending its range of action even further. Therefore, the expression of DAX-1 could be a critical determinant of nuclear function, and it is not surprising that DAX-1 expression is dynamically regulated. For example, its expression in the developing gonads appears to diminish at certain developmental periods (28, 59). Furthermore, expression of DAX-1 in the testicular Sertoli cell is cyclical and is inhibited by cyclic AMP (60).

The ability of DAX-1 to repress transcription when bound directly or indirectly to DNA indicates that the mechanism of repression utilized by DAX-1 may be conserved in either case. This is further supported by the fact that all known AHC mutations of DAX-1 mutate or delete the LBD, and as a group, these mutations appear to disrupt the ability to interact with N-CoR. Thus, DAX-1 is likely to play important roles in nuclear receptor corepressor signaling. However, it is conceivable that DAX-1 uses different repressive mechanisms in particular cellular and/or promoter contexts. Alternatively, DAX-1 could under certain conditions activate transcription, because it does harbor a conserved AF-2-AH. Therefore, DAX-1 probably has functions outside the realm of SF-1 regulation, and their delineation will be important for our understanding of the role of DAX-1 in organogenesis, steroidogenesis, and reproductive function.

    ACKNOWLEDGMENTS

We thank M. Lazar, A. Hörlein, J. D. Chen, and P. Webb for providing plasmids used in this study. We thank S. Audrain for sequencing constructs. Finally, we thank J. Svaren and T. Wilson for critical review of the manuscript.

This work was supported by a grant from the National Cancer Institute (PO1-49712-07) (J.M.), an MSTP training grant (P.A.C.), and NIH grant HD-34110 (Y.S.).

    FOOTNOTES

* Corresponding author. Mailing address: Department of Pathology and Internal Medicine, Washington University School of Medicine, 660 S. Euclid Ave., Box 8118, St. Louis, MO 63110. Phone: (314) 362-4650. Fax: (314) 362-8756. E-mail: jeff{at}milbrandt.wustl.edu.

    REFERENCES
Top
Abstract
Introduction
Materials & Methods
Results
Discussion
References

1. Alland, L., R. Muhle, J. Hou, J. J. Potes, L. Chin, N. Schrieber-Agus, and R. DePinho. 1997. Role for N-CoR and histone deacetylase in Sin3-mediated transcriptional repression. Nature 387:49-55[Medline].
2. Bae, D. S., M. L. Schaefer, B. W. Partan, and L. Muglia. 1996. Characterization of the mouse DAX-1 gene reveals evolutionary conservation of a unique amino-terminal motif and widespread expression in mouse tissue. Endocrinology 137:3921-3927[Abstract].
3. Bakke, M., and J. Lund. 1995. Mutually exclusive interactions of two nuclear orphan receptors determine activity of a cAMP-responsive sequence in the bovine CYP17 gene. Mol. Endocrinol. 9:327-339[Abstract/Free Full Text].
4. Baniahmad, A., X. Leng, T. P. Burris, S. Y. Tsai, M.-J. Tsai, and B. W. O'Malley. 1995. The tau 4 activation domain of the thyroid hormone receptor is required for release of a putative corepressor(s) necessary for transcriptional silencing. Mol. Cell. Biol. 15:76-86[Abstract].
5. Barnes, W. M. 1994. PCR amplification of up to 35-kb DNA with high fidelity and high yield from lambda bacteriophage templates. Proc. Natl. Acad. Sci. USA 91:2216-2220[Abstract/Free Full Text].
6. Barnhart, K. M., and P. L. Mellon. 1994. The orphan nuclear receptor, SF-1, regulates the glycoprotein hormone alpha-subunit gene in pituitary gonadotropes. Mol. Endocrinol. 8:878-885[Abstract/Free Full Text].
7. Beato, M., P. Herrlich, and G. Shutz. 1995. Steroid hormone receptors: many actors in search of a plot. Cell 83:851-857[Medline].
8. Bourguet, W., M. Ruff, P. Chambon, H. Gronemeyer, and D. Moras. 1995. Crystal structure of the ligand-binding domain of the human nuclear receptor RXR-alpha . Nature 375:377-382[Medline].
9. Brewer, C. B. 1994. Cytomegalovirus plasmid vectors for permanent lines of polarized epithelial cells. Methods Cell Biol. 43:233-245.
10. Cammas, F. M., G. D. Pullinger, S. Barker, and A. J. L. Clark. 1997. The mouse adrenocorticotropin receptor gene: cloning and characterization of its promoter and evidence for a role for the orphan nuclear receptor steroidogenic factor 1. Mol. Endocrinol. 11:867-876[Abstract/Free Full Text].
11. Caron, K. M., Y. Ikeda, S.-C. Soo, D. M. Stocco, K. L. Parker, and B. J. Clark. 1997. Characterization of the promoter region of the mouse gene encoding the steroidogenic acute regulatory protein. Mol. Endocrinol. 11:138-147[Abstract/Free Full Text].
12. Chen, J. D., and R. M. Evans. 1995. A transcriptional co-repressor that interacts with nuclear hormone receptors. Nature 377:454-457[Medline].
13. Clemens, J. W., D. S. Lala, K. L. Parker, and J. S. Richards. 1994. Steroidogenic factor-1 binding and transcriptional activity of the cholesterol side-chain cleavage promoter in rat granulosa cells. Endocrinology 134:1499-1508[Abstract/Free Full Text].
14. Crawford, P., Y. Sadovsky, and J. Milbrandt. 1997. Nuclear receptor steroidogenic factor-1 directs embryonic stem cells toward the steroidogenic lineage. Mol. Cell. Biol. 17:3997-4006[Abstract].
15. Crawford, P. A., J. A. Polish, G. Ganpule, and Y. Sadovsky. 1997. The activation function-2 domain of steroidogenic factor-1 is required, but not sufficient for potentiation by SRC-1. Mol. Endocrinol. 11:1626-1635[Abstract/Free Full Text].
16. Danielian, P. S., R. White, J. A. Lees, and M. G. Parker. 1992. Identification of a conserved region required for hormone dependent transcriptional activation by steroid hormone receptors. EMBO J. 11:1025-1033[Medline].
17. Giuili, G., W.-H. Shen, and H. A. Ingraham. 1997. The nuclear receptor SF-1 mediates sexually dimorphic expression of Mullerian inhibiting substance, in vivo. Development 124:1799-1807[Abstract].
18. Halvorson, L. M., U. B. Kaiser, and W. W. Chin. 1996. Stimulation of luteinzing hormone beta gene promoter activity by the orphan receptor, steroidogenic factor-1. J. Biol. Chem. 271:6645-6650[Abstract/Free Full Text].
19. Harding, H. P., G. B. Atkins, A. B. Jaffe, W. J. Seo, and M. A. Lazar. 1997. Transcriptional activation and repression by RORalpha , and orphan nuclear receptor required for cerebellar development. Mol. Endocrinol. 11:1737-1746[Abstract/Free Full Text].
20. Heizel, T., R. Lavinsky, T.-M. Mullen, M. Soderstrom, C. Laherty, J. Torchia, W.-M. Yang, G. Brard, S. Ngo, J. Davie, E. Seto, R. Eisenmann, D. Rose, C. Glass, and M. Rosenfeld. 1997. A complex containing N-CoR, mSin3 and histone deacetylase mediates transcriptional repression. Nature 387:43-48[Medline].
21. Honda, S.-I., K.-I. Morohashi, M. Nomura, H. Takeya, M. Kitajima, and T. Omura. 1993. Ad4BP regulating steroidogenic P450 gene is a member of steroid hormone receptor superfamily. J. Biol. Chem. 268:7494-7502[Abstract/Free Full Text].
22. Horlein, A. J., A. M. Naar, T. Heinzel, J. Torchia, B. Gloss, R. Kurokawa, A. Ryan, Y. Kamei, M. Soderstrom, C. K. Glass, and M. G. Rosenfeld. 1995. Ligand-independent repression by the thyroid hormone receptor mediated by a nuclear receptor co-repressor. Nature 377:397-404[Medline].
23. Horwitz, K. B., T. A. Jackson, D. L. Bain, J. K. Richer, G. S. Takimoto, and L. Tung. 1996. Nuclear receptor coactivators and corepressors. Mol. Endocrinol. 10:1167-1177[Abstract/Free Full Text].
24. Hu, Z., L. Zhuang, X. Guan, J. Meng, and M. L. Dufau. 1997. Steroidogenic factor-1 is an essential transcriptional activator for gonad-specific expression of promoter I of the rat prolactin receptor gene. J. Biol. Chem. 272:14263-14271[Abstract/Free Full Text].
25. Ikeda, Y., D. S. Lala, X. Luo, E. Kim, M. P. Moisan, and K. L. Parker. 1993. Characterization of the mouse FTZ-F1 gene, which encodes a key regulator of steroid hydroxylase gene expression. Mol. Endocrinol. 7:852-860[Abstract/Free Full Text].
26. Ikeda, Y., X. Luo, R. Abbud, J. H. Nilson, and K. L. Parker. 1995. The nuclear receptor SF-1 is essential for the formation of the ventromedial hypothalamic nucleus. Mol. Endocrinol. 9:478-486[Abstract/Free Full Text].
27. Ikeda, Y., W.-H. Shen, H. A. Ingraham, and K. L. Parker. 1994. Developmental expression of mouse steroidogenic factor-1, an essential regulator of the steroid hydroxylases. Mol. Endocrinol. 8:654-662[Abstract/Free Full Text].
28. Ikeda, Y., A. Swain, T. J. Weber, K. E. Hentges, E. Zanaria, E. Lalli, K. T. Tamai, P. Sassone-Corsi, R. Lovell-Badge, G. Camerino, and K. L. Parker. 1996. Steroidogenic factor 1 and Dax-1 colocalize in multiple cell lineages: potential links in endocrine development. Mol. Endocrinol. 10:1262-1272.
29. Ingraham, H. A., D. S. Lala, Y. Ikeda, X. Luo, W.-H. Shen, M. W. Nachtigal, R. Abbud, J. H. Nilson, and K. L. Parker. 1994. The nuclear receptor SF-1 acts at multiple levels of the reproductive axis. Genes Dev. 8:2302-2312[Abstract/Free Full Text].
30. Ito, M., R. Yu, and J. L. Jameson. 1997. DAX-1 inhibits SF-1-mediated transactivation via a carboxy-terminal domain that is deleted in adrenal hypoplasia congenita. Mol. Cell. Biol. 17:1476-1483[Abstract].
31. Kastner, P., M. Mark, and P. Chambon. 1995. Nonsteroid nuclear receptors: what are genetic studies telling us about their role in real life? Cell 83:859-869[Medline].
32. Keri, R. A., and J. H. Nilson. 1996. A steroidogenic factor-1 binding site is required for activity of the luteinizing hormone beta subunit promoter in gonadotropes of transgenic mice. J. Biol. Chem. 271:10782-10785[Abstract/Free Full Text].
33. Kurokawa, R., M. Soderstrom, A. Horlein, S. Halachmi, M. Brown, M. G. Rosenfeld, and C. K. Glass. 1995. Polarity-specific activities of retinoic acid receptors determined by a co-repressor. Nature 377:451-454[Medline].
34. Lala, D. S., P. M. Syka, S. B. Lazarchik, D. J. Mangelsdorf, K. L. Parker, and R. A. Heyman. 1997. Activation of the orphan nuclear receptor steroidogenic factor 1 by oxysterols. Proc. Natl. Acad. Sci. USA 94:4895-4900[Abstract/Free Full Text].
35. Lee, S. L., Y. Sadovsky, A. H. Swirnoff, J. A. Polish, P. Goda, G. Gavrilina, and J. Milbrandt. 1996. Luteinizing hormone deficiency and female infertility in mice lacking the transcription factor NGFI-A (Egr-1). Science 273:1219-1222[Abstract].
36. Leers-Sucheta, S., K. Morohashi, J. Mason, and M. Melner. 1997. Synergistic activation of the human type II 3b-HSD/D5-D4 isomerase promoter by the transcription factor SF-1/Ad4BP and phorbol ester. J. Biol. Chem. 272:7960-7967[Abstract/Free Full Text].
37. Leng, X., J. Blanco, S. Y. Tsai, K. Ozato, B. W. O'Malley, and M.-J. Tsai. 1995. Mouse retinoid X receptor contains a separable ligand-binding and transactivation domain in its E region. Mol. Cell. Biol. 15:255-263[Abstract].
38. Luo, X., Y. Ikeda, and K. L. Parker. 1994. A cell-specific nuclear receptor is essential for adrenal and gonadal development and sexual differentiation. Cell 77:481-490[Medline].
39. Mangelsdorf, D. J., and R. M. Evans. 1995. The RXR heterodimers and orphan receptors. Cell 83:841-850[Medline].
40. Mangelsdorf, D. J., C. Thummel, M. Beato, P. Herrlich, G. Schutz, K. Umesono, B. Blumberg, P. Kastner, M. Mark, P. Chambon, and R. M. Evans. 1995. The nuclear receptor superfamily: the second decade. Cell 83:835-839[Medline].
41. Morohashi, K., S. Honda, Y. Inomata, H. Handa, and T. Omura. 1992. A common trans-acting factor, Ad4-binding protein, to the promoters of steroidogenic P-450s. J. Biol. Chem. 267:17913-17919[Abstract/Free Full Text].
42. Morohashi, K.-I., H. Iida, M. Nomura, O. Hatano, S.-I. Honda, T. Tsukiyama, O. Niwa, T. Hara, A. Takakusu, Y. Shibata, and T. Omura. 1994. Functional difference between Ad4BP and ELP, and their distributions in steroidogenic tissues. Mol. Endocrinol. 8:643-653[Abstract/Free Full Text].
43. Morohashi, K.-I., U. M. Zanger, S.-I. Honda, M. Hara, M. R. Waterman, and T. Omura. 1993. Activation of CYP11A and CYP11B gene promoters by the steroidogenic cell-specific transcription factor, Ad4BP. Mol. Endocrinol. 7:1196-1204[Abstract/Free Full Text].
44. Muscatelli, F., T. M. Strom, A. P. Walker, E. Zanaria, D. Recan, A. Meindi, B. Bardoni, S. Guioli, G. Zehtner, W. Rabl, H. P. Schwarz, J.-C. Kaplan, G. Camerino, T. Meitinger, and A. P. Monaco. 1994. Mutations in the DAX-1 gene give rise to both adrenal hypoplasia congenita and hypogonadotropic hypogonadism. Nature 372:672-676[Medline].
45. Nagy, L., H.-Y. Kao, D. Chakravarti, R. J. Lin, C. A. Hassig, D. E. Ayer, S. L. Scrieber, and R. M. Evans. 1997. Nuclear receptor repression mediated by a complex containing SMRT, mSin3A, and histone deacetylase. Cell 89:373-380[Medline].
46. Perlmann, T., K. Umesono, P. N. Rangarajan, B. M. Forman, and R. M. Evans. 1996. Two distinct dimerization interfaces differentially modulate target gene specificity of nuclear hormone receptors. Mol. Endocrinol. 10:958-966[Abstract/Free Full Text].
47. Preiss, A., U. B. Rosenberg, A. Kienlin, E. Seifert, and H. Jackle. 1985. Molecular genetics of Kruppel, a gene required for segmentation of the Drosophila embryo. Nature 313:27-32[Medline].
48. Renaud, J.-P., N. Rochel, M. Ruff, V. Vivat, P. Chambon, H. Gronemeyer, and D. Moras. 1995. Crystal structure of the RARgamma ligand-binding domain bound to all-trans retinoic acid. Nature 378:681-689[Medline].
49. Rice, D. A., A. R. Mouw, and K. L. Parker. 1991. A shared promoter element regulates the expression of three steroidogenic enzymes. Mol. Endocrinol. 5:1552-1561[Abstract/Free Full Text].
49a. Sadovsky, Y. Unpublished data.
50. Sadovsky, Y., and P. A. Crawford. 1998. Developmental and physiologic roles of the nuclear receptor steroidogenic factor 1 in the reproductive system. J. Soc. Gynecol. Invest. 5:6-12. [Medline]
51. Sadovsky, Y., P. A. Crawford, K. G. Woodson, J. A. Polish, M. A. Clements, L. M. Tourtellotte, K. Simburger, and J. Milbrandt. 1995. Mice deficient in the orphan receptor steroidogenic factor 1 lack adrenal glands and gonads but express P450 side-chain-cleavage enzyme in the placenta, and have normal embryonic serum levels of corticosteroids. Proc. Natl. Acad. Sci. USA 92:10939-10943[Abstract/Free Full Text].
52. Sadowski, I., and M. Ptashne. 1989. A vector for expressing GAL4(1-147) fusions in mammalian cells. Nucleic Acids Res. 17:7539[Free Full Text].
53. Sauer, F., and H. Jackle. 1993. Dimerization and the control of transcription by Kruppel. Nature 364:454-457[Medline].
54. Seol, W., M. J. Mahon, Y. K. Lee, and D. D. Moore. 1996. Two receptor interacting domains in the nuclear hormone receptor corepressor RIP13/N-CoR. Mol. Endocrinol. 10:1646-1655[Abstract/Free Full Text].
55. Shen, W.-H., C. C. D. Moore, Y. Ikeda, K. L. Parker, and H. A. Ingraham. 1994. Nuclear receptor steroidogenic factor 1 regulates the Mullerian inhibiting substance gene: a link to the sex determination cascade. Cell 77:651-661[Medline].
56. Shibata, H., Z. Nawaz, S. Y. Tsai, B. W. O'Malley, and M. J. Tsai. 1997. Gene silencing by chicken ovalbumin upstream promoter-transcription factor I (COUP-TFI) is mediated by transcriptional corepressors, nuclear receptor-corepressor (N-CoR) and silencing mediator for retinoic acid receptor and thyroid hormone receptor (SMRT). Mol. Endocrinol. 11:714-724[Abstract/Free Full Text].
57. Shinoda, K., H. Lei, H. Yoshii, M. Nomura, M. Nagano, H. Shiba, H. Sasaki, Y. Osawa, Y. Ninomiya, O. Niwa, K.-I. Morohashi, and E. Li. 1995. Developmental defects of the ventromedial hypothalamic nucleus and pituitary gonadotroph in the Ftz-F1 disrupted mice. Dev. Dyn. 204:22-29[Medline].
58. Sugawara, T., J. A. Holt, M. Kiriakidou, and J. F. Strauss, III. 1996. Steroidogenic factor 1-dependent promoter activity of the human steroidogenic acute regulatory protein (StAR) gene. Biochemistry 35:9052-9059[Medline].
59. Swain, A., E. Zanaria, A. Hacker, R. Lovell-Badge, and G. Camerino. 1996. Mouse Dax1 expression is consistent with a role in sex determination as well as in adrenal and hypothalamus function. Nat. Genet. 12:404-409[Medline].
60. Tamai, K. T., L. Monaco, T. P. Alastalo, E. Lalli, M. Parvinen, and P. Sassone-Corsi. 1996. Hormonal and developmental regulation of DAX-1 expression in Sertoli cells. Mol. Endocrinol. 10:1561-1569[Abstract/Free Full Text].
61. Wagner, R. L., J. W. Apriletti, M. E. McGrath, B. L. West, J. D. Baxter, and R. J. Fletterick. 1995. A structural role for hormone in the thyroid hormone receptor. Nature 378:690-697[Medline].
62. Wehrenberg, U., R. Ivell, M. Jansen, S. von Goedecke, and N. Walther. 1994. Two orphan receptors binding to a common site are involved in the regulation of the oxytocin gene in the bovine ovary. Proc. Natl. Acad. Sci. USA 91:1440-1444[Abstract/Free Full Text].
63. Wilson, T. E., T. J. Fahrner, and J. Milbrandt. 1993. The orphan receptors NGFI-B and steroidogenic factor 1 establish monomer binding as a third paradigm of nuclear receptor-DNA interaction. Mol. Cell. Biol. 13:5794-5804[Abstract/Free Full Text].
64. Wilson, T. E., A. R. Mouw, C. A. Weaver, J. Milbrandt, and K. L. Parker. 1993. The orphan nuclear receptor NGFI-B regulates expression of the gene encoding steroid 21-hydroxylase. Mol. Cell. Biol. 13:861-868[Abstract/Free Full Text].
65. Woodson, K. G., P. A. Crawford, Y. Sadovsky, and J. Milbrandt. 1997. Characterization of the promoter of SF-1, an orphan nuclear receptor required for adrenal and gonadal development. Mol. Endocrinol. 11:117-126[Abstract/Free Full Text].
66. Zamir, I., H. P. Harding, G. B. Atkins, A. Hörlein, C. K. Glass, M. G. Rosenfeld, and M. A. Lazar. 1996. A nuclear hormone receptor corepressor mediates transcriptional silencing by receptors with distinct repression domains. Mol. Cell. Biol. 16:5458-5465[Abstract].
67. Zamir, I., J. Zhang, and M. A. Lazar. 1997. Stoichiometric and steric principles governing repression by nuclear hormone receptors. Genes Dev. 11:835-846[Abstract/Free Full Text].
68. Zanaria, E., F. Muscatelli, B. Bardoni, T. M. Strom, S. Guioli, W. Guo, E. Lalli, C. Moser, A. P. Walker, E. R. B. McCabe, T. Meitinger, A. P. Monaco, P. Sassone-Corsi, and G. Camerino. 1994. An unusual member of the nuclear hormone receptor superfamily responsible for X-linked adrenal hypoplasia congenita. Nature 372:635-641[Medline].
69. Zazopoulos, E., E. Lalli, D. M. Stocco, and P. Sassone-Corsi. 1997. DNA binding and transcriptional repression by DAX-1 blocks steroidogenesis. Nature 390:311-315[Medline].
70. Zhang, P., and S. H. Mellon. 1996. The orphan nuclear receptor steroidogenic factor-1 regulates the cAMP-mediated transcriptional activation of rat cytochrome P450c17 (17a-hydroxylase/c17-20 lyase). Mol. Endocrinol. 10:147-158[Abstract/Free Full Text].

Mol Cell Biol, May 1998, p. 2949-2956, Vol. 18, No. 5
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  • MacLaughlin, D. T., Donahoe, P. K. (2004). Sex Determination and Differentiation. NEJM 350: 367-378 [Full Text]  
  • Fan, W., Yanase, T., Wu, Y., Kawate, H., Saitoh, M., Oba, K., Nomura, M., Okabe, T., Goto, K., Yanagisawa, J., Kato, S., Takayanagi, R., Nawata, H. (2004). Protein Kinase A Potentiates Adrenal 4 Binding Protein/Steroidogenic Factor 1 Transactivation by Reintegrating the Subcellular Dynamic Interactions of the Nuclear Receptor with Its Cofactors, General Control Nonderepressed-5/Transformation/ Transcription Domain-Associated Protein, and Suppressor, Dosage-Sensitive Sex Reversal-1: a Laser Confocal Imaging Study in Living KGN Cells. Mol. Endocrinol. 18: 127-141 [Abstract] [Full Text]  
  • Borud, B., Mellgren, G., Lund, J., Bakke, M. (2003). Cloning and Characterization of a Novel Zinc Finger Protein that Modulates the Transcriptional Activity of Nuclear Receptors. Mol. Endocrinol. 17: 2303-2319 [Abstract] [Full Text]  
  • Liu, Y.-W., Gao, W., Teh, H.-L., Tan, J.-H., Chan, W.-K. (2003). Prox1 Is a Novel Coregulator of Ff1b and Is Involved in the Embryonic Development of the Zebra Fish Interrenal Primordium. Mol. Cell. Biol. 23: 7243-7255 [Abstract] [Full Text]  
  • Hong, C. Y., Park, J. H., Seo, K. H., Kim, J.-M., Im, S. Y., Lee, J. W., Choi, H.-S., Lee, K. (2003). Expression of MIS in the Testis Is Downregulated by Tumor Necrosis Factor Alpha through the Negative Regulation of SF-1 Transactivation by NF-{kappa}B. Mol. Cell. Biol. 23: 6000-6012 [Abstract] [Full Text]  
  • Ourlin, J. C., Lasserre, F., Pineau, T., Fabre, J. M., Sa-Cunha, A., Maurel, P., Vilarem, M.-J., Pascussi, J. M. (2003). The Small Heterodimer Partner Interacts with the Pregnane X Receptor and Represses Its Transcriptional Activity. Mol. Endocrinol. 17: 1693-1703 [Abstract] [Full Text]  
  • Agoulnik, I. U., Krause, W. C., Bingman, W. E. III, Rahman, H. T., Amrikachi, M., Ayala, G. E., Weigel, N. L. (2003). Repressors of Androgen and Progesterone Receptor Action. J. Biol. Chem. 278: 31136-31148 [Abstract] [Full Text]  
  • Lalli, E., Sassone-Corsi, P. (2003). DAX-1, an Unusual Orphan Receptor at the Crossroads of Steroidogenic Function and Sexual Differentiation. Mol. Endocrinol. 17: 1445-1453 [Abstract] [Full Text]  
  • Gummow, B. M., Winnay, J. N., Hammer, G. D. (2003). Convergence of Wnt Signaling and Steroidogenic Factor-1 (SF-1) on Transcription of the Rat Inhibin {alpha} Gene. J. Biol. Chem. 278: 26572-26579 [Abstract] [Full Text]  
  • Sato, Y., Suzuki, T., Hidaka, K., Sato, H., Ito, K., Ito, S., Sasano, H. (2003). Immunolocalization of Nuclear Transcription Factors, DAX-1 and COUP-TF II, in the Normal Human Ovary: Correlation with Adrenal 4 Binding Protein/ Steroidogenic Factor-1 Immunolocalization during the Menstrual Cycle. J. Clin. Endocrinol. Metab. 88: 3415-3420 [Abstract] [Full Text]  
  • Ghosh, A. K., Steele, R., Ray, R. B. (2003). Modulation of Human Luteinizing Hormone {beta} Gene Transcription by MIP-2A. J. Biol. Chem. 278: 24033-24038 [Abstract] [Full Text]  
  • Kawajiri, K., Ikuta, T., Suzuki, T., Kusaka, M., Muramatsu, M., Fujieda, K., Tachibana, M., Morohashi, K.-i. (2003). Role of the LXXLL-Motif and Activation Function 2 Domain in Subcellular Localization of Dax-1 (Dosage-Sensitive Sex Reversal-Adrenal Hypoplasia Congenita Critical Region on the X Chromosome, Gene 1). Mol. Endocrinol. 17: 994-1004 [Abstract] [Full Text]  
  • Yazawa, T., Mizutani, T., Yamada, K., Kawata, H., Sekiguchi, T., Yoshino, M., Kajitani, T., Shou, Z., Miyamoto, K. (2003). Involvement of Cyclic Adenosine 5'-Monophosphate Response Element-Binding Protein, Steroidogenic Factor 1, and Dax-1 in the Regulation of Gonadotropin-Inducible Ovarian Transcription Factor 1 Gene Expression by Follicle-Stimulating Hormone in Ovarian Granulosa Cells. Endocrinology 144: 1920-1930 [Abstract] [Full Text]  
  • Sadie, H., Styger, G., Hapgood, J. (2003). Expression of the Mouse Gonadotropin-Releasing Hormone Receptor Gene in {alpha}T3-1 Gonadotrope Cells Is Stimulated by Cyclic 3',5'-Adenosine Monophosphate and Protein Kinase A, and Is Modulated by Steroidogenic Factor-1 and Nur77. Endocrinology 144: 1958-1971 [Abstract] [Full Text]  
  • Mizusaki, H., Kawabe, K., Mukai, T., Ariyoshi, E., Kasahara, M., Yoshioka, H., Swain, A., Morohashi, K.-i. (2003). Dax-1 (Dosage-Sensitive Sex Reversal-Adrenal Hypoplasia Congenita Critical Region on the X Chromosome, Gene 1) Gene Transcription Is Regulated by Wnt4 in the Female Developing Gonad. Mol. Endocrinol. 17: 507-519 [Abstract] [Full Text]  
  • Brown, P., Scobie, G. A., Townsend, J., Bayne, R. A. L., Seckl, J. R., Saunders, P. T. K., Anderson, R. A. (2003). Identification of a Novel Missense Mutation That Is as Damaging to DAX-1 Repressor Function as a Nonsense Mutation. J. Clin. Endocrinol. Metab. 88: 1341-1349 [Abstract] [Full Text]  
  • Liao, G., Chen, L.-Y., Zhang, A., Godavarthy, A., Xia, F., Ghosh, J. C., Li, H., Chen, J. D. (2003). Regulation of Androgen Receptor Activity by the Nuclear Receptor Corepressor SMRT. J. Biol. Chem. 278: 5052-5061 [Abstract] [Full Text]  
  • Suzuki, T., Kasahara, M., Yoshioka, H., Morohashi, K.-i., Umesono, K. (2003). LXXLL-Related Motifs in Dax-1 Have Target Specificity for the Orphan Nuclear Receptors Ad4BP/SF-1 and LRH-1. Mol. Cell. Biol. 23: 238-249 [Abstract] [Full Text]  
  • Yan, X., Mouillet, J.-F., Ou, Q., Sadovsky, Y. (2003). A Novel Domain within the DEAD-Box Protein DP103 Is Essential for Transcriptional Repression and Helicase Activity. Mol. Cell. Biol. 23: 414-423 [Abstract] [Full Text]  
  • Wei, X., Sasaki, M., Huang, H., Dawson, V. L., Dawson, T. M. (2002). The Orphan Nuclear Receptor, Steroidogenic Factor 1, Regulates Neuronal Nitric Oxide Synthase Gene Expression in Pituitary Gonadotropes. Mol. Endocrinol. 16: 2828-2839 [Abstract] [Full Text]  
  • Osman, H., Murigande, C., Nadakal, A., Capponi, A. M. (2002). Repression of DAX-1 and Induction of SF-1 Expression. TWO MECHANISMS CONTRIBUTING TO THE ACTIVATION OF ALDOSTERONE BIOSYNTHESIS IN ADRENAL GLOMERULOSA CELLS. J. Biol. Chem. 277: 41259-41267 [Abstract] [Full Text]  
  • Gizard, F., Lavallee, B., DeWitte, F., Teissier, E., Staels, B., Hum, D. W. (2002). The Transcriptional Regulating Protein of 132 kDa (TReP-132) Enhances P450scc Gene Transcription through Interaction with Steroidogenic Factor-1 in Human Adrenal Cells. J. Biol. Chem. 277: 39144-39155 [Abstract] [Full Text]  
  • Moraitis, A. N., Giguere, V., Thompson, C. C. (2002). Novel Mechanism of Nuclear Receptor Corepressor Interaction Dictated by Activation Function 2 Helix Determinants. Mol. Cell. Biol. 22: 6831-6841 [Abstract] [Full Text]  
  • Kajiyama, Y., Tian, J., Locker, J. (2002). Regulation of {alpha}-Fetoprotein Expression by Nkx2.8. Mol. Cell. Biol. 22: 6122-6130 [Abstract] [Full Text]  
  • Brendel, C., Schoonjans, K., Botrugno, O. A., Treuter, E., Auwerx, J. (2002). The Small Heterodimer Partner Interacts with the Liver X Receptor {alpha} and Represses Its Transcriptional Activity. Mol. Endocrinol. 16: 2065-2076 [Abstract] [Full Text]  
  • Lehmann, S. G., Lalli, E., Sassone-Corsi, P. (2002). From the Cover: X-linked adrenal hypoplasia congenita is caused by abnormal nuclear localization of the DAX-1 protein. Proc. Natl. Acad. Sci. USA 99: 8225-8230 [Abstract] [Full Text]  
  • Seminara, S. B., Crowley, W. F. Jr. (2002). Genetic Approaches to Unraveling Reproductive Disorders: Examples of Bedside to Bench Research in the Genomic Era. Endocr. Rev. 23: 382-392 [Abstract] [Full Text]  
  • Holter, E., Kotaja, N., Makela, S., Strauss, L., Kietz, S., Janne, O. A., Gustafsson, J.-A., Palvimo, J. J., Treuter, E. (2002). Inhibition of Androgen Receptor (AR) Function by the Reproductive Orphan Nuclear Receptor DAX-1. Mol. Endocrinol. 16: 515-528 [Abstract] [Full Text]  
  • Koskimies, P., Levallet, J., Sipila, P., Huhtaniemi, I., Poutanen, M. (2002). Murine Relaxin-Like Factor Promoter: Functional Characterization and Regulation by Transcription Factors Steroidogenic Factor 1 and DAX-1. Endocrinology 143: 909-919 [Abstract] [Full Text]  
  • Jepsen, K., Rosenfeld, M. G. (2002). Biological roles and mechanistic actions of co-repressor complexes. J. Cell Sci. 115: 689-698 [Abstract] [Full Text]  
  • Marimuthu, A., Feng, W., Tagami, T., Nguyen, H., Jameson, J. L., Fletterick, R. J., Baxter, J. D., West, B. L. (2002). TR Surfaces and Conformations Required to Bind Nuclear Receptor Corepressor. Mol. Endocrinol. 16: 271-286 [Abstract] [Full Text]  
  • Babu, P. S., Bavers, D. L., Beuschlein, F., Shah, S., Jeffs, B., Jameson, J. L., Hammer, G. D. (2002). Interaction Between Dax-1 and Steroidogenic Factor-1 in Vivo: Increased Adrenal Responsiveness to ACTH in the Absence of Dax-1. Endocrinology 143: 665-673 [Abstract] [Full Text]  
  • McCabe, E. R. B. (2002). Vulnerability within a Robust Complex System--DAX-1 Mutations and Steroidogenic Axis Development. J. Clin. Endocrinol. Metab. 87: 41-43 [Abstract] [Full Text]  
  • Parker, K. L., Rice, D. A., Lala, D. S., Ikeda, Y., Luo, X., Wong, M., Bakke, M., Zhao, L., Frigeri, C., Hanley, N. A., Stallings, N., Schimmer, B. P. (2002). Steroidogenic Factor 1: an Essential Mediator of Endocrine Development. Recent Prog Horm Res 57: 19-36 [Abstract] [Full Text]  
  • Lopez, D., Shea-Eaton, W., Sanchez, M. D., McLean, M. P. (2001). DAX-1 Represses the High-Density Lipoprotein Receptor Through Interaction with Positive Regulators Sterol Regulatory Element-Binding Protein-1a and Steroidogenic Factor-1. Endocrinology 142: 5097-5106 [Abstract] [Full Text]  
  • Yuan, X., Lu, M. L., Li, T., Balk, S. P. (2001). SRY Interacts with and Negatively Regulates Androgen Receptor Transcriptional Activity. J. Biol. Chem. 276: 46647-46654 [Abstract] [Full Text]  
  • Nilsson, S., Makela, S., Treuter, E., Tujague, M., Thomsen, J., Andersson, G., Enmark, E., Pettersson, K., Warner, M., Gustafsson, J.-A. (2001). Mechanisms of Estrogen Action. Physiol. Rev. 81: 1535-1565 [Abstract] [Full Text]  
  • Sugawara, T., Abe, S., Sakuragi, N., Fujimoto, Y., Nomura, E., Fujieda, K., Saito, M., Fujimoto, S. (2001). RIP 140 Modulates Transcription of the Steroidogenic Acute Regulatory Protein Gene through Interactions with Both SF-1 and DAX-1. Endocrinology 142: 3570-3577 [Abstract] [Full Text]  
  • Achermann, J. C., Ito, M., Silverman, B. L., Habiby, R. L., Pang, S., Rosler, A., Jameson, J. L. (2001). Missense Mutations Cluster within the Carboxyl-Terminal Region of DAX-1 and Impair Transcriptional Repression. J. Clin. Endocrinol. Metab. 86: 3171-3175 [Abstract] [Full Text]  
  • Aranda, A., Pascual, A. (2001). Nuclear Hormone Receptors and Gene Expression. Physiol. Rev. 81: 1269-1304 [Abstract] [Full Text]  
  • Wang, Z. J., Jeffs, B., Ito, M., Achermann, J. C., Yu, R. N., Hales, D. B., Jameson, J. L. (2001). Aromatase (Cyp19) expression is up-regulated by targeted disruption of Dax1. Proc. Natl. Acad. Sci. USA 10.1073/pnas.141543298v1 [Abstract] [Full Text]  
  • Aylwin, S. J. B., Welch, J. P., Davey, C. L., Geddes, J. F., Wood, D. F., Besser, G. M., Grossman, A. B., Monson, J. P., Burrin, J. M. (2001). The Relationship between Steroidogenic Factor 1 and DAX-1 Expression and in Vitro Gonadotropin Secretion in Human Pituitary Adenomas. J. Clin. Endocrinol. Metab. 86: 2476-2483 [Abstract] [Full Text]  
  • Calvo, R. M., Asunción, M., Tellería, D., Sancho, J., San Millán, J. L., Escobar-Morreale, H. F. (2001). Screening for Mutations in the Steroidogenic Acute Regulatory Protein and Steroidogenic Factor-1 Genes, and in CYP11A and Dosage-Sensitive Sex Reversal-Adrenal Hypoplasia Gene on the X Chromosome, Gene-1 (DAX-1), in Hyperandrogenic Hirsute Women. J. Clin. Endocrinol. Metab. 86: 1746-1749 [Abstract] [Full Text]  
  • Tremblay, J. J., Viger, R. S. (2001). Nuclear Receptor Dax-1 Represses the Transcriptional Cooperation Between GATA-4 and SF-1 in Sertoli Cells. Biol. Reprod. 64: 1191-1199 [Abstract] [Full Text]  
  • Giannoukos, G., Szapary, D., Smith, C. L., Meeker, J. E. W., Simons, S. S. Jr. (2001). New Antiprogestins with Partial Agonist Activity: Potential Selective Progesterone Receptor Modulators (SPRMs) and Probes for Receptor- and Coregulator-Induced Changes in Progesterone Receptor Induction Properties. Mol. Endocrinol. 15: 255-270 [Abstract] [Full Text]  
  • Hanley, N. A., Rainey, W. E., Wilson, D. I., Ball, S. G., Parker, K. L. (2001). Expression Profiles of SF-1, DAX1, and CYP17 in the Human Fetal Adrenal Gland: Potential Interactions in Gene Regulation. Mol. Endocrinol. 15: 57-68 [Abstract] [Full Text]  
  • Ou, Q., Mouillet, J.-F., Yan, X., Dorn, C., Crawford, P. A., Sadovsky, Y. (2001). The DEAD Box Protein DP103 Is a Regulator of Steroidogenic Factor-1. Mol. Endocrinol. 15: 69-79 [Abstract] [Full Text]  
  • Levallet, J., Koskimies, P., Rahman, N., Huhtaniemi, I. (2001). The Promoter of Murine Follicle-Stimulating Hormone Receptor: Functional Characterization and Regulation by Transcription Factor Steroidogenic Factor 1. Mol. Endocrinol. 15: 80-92 [Abstract] [Full Text]  
  • Aigueperse, C., Val, P., Pacot, C., Darne, C., Lalli, E., Sassone-Corsi, P., Veyssiere, G., Jean, C., Martinez, A. (2001). SF-1 (Steroidogenic Factor-1), C/EBP{beta} (CCAAT/Enhancer Binding Protein), and Ubiquitous Transcription Factors NF1 (Nuclear Factor 1) and Sp1 (Selective Promoter Factor 1) Are Required for Regulation of the Mouse Aldose Reductase-Like Gene (AKR1B7) Expression in Adrenocortical Cells. Mol. Endocrinol. 15: 93-111 [Abstract] [Full Text]  
  • Bland, M. L., Jamieson, C. A. M., Akana, S. F., Bornstein, S. R., Eisenhofer, G., Dallman, M. F., Ingraham, H. A. (2000). Haploinsufficiency of steroidogenic factor-1 in mice disrupts adrenal development leading to an impaired stress response. Proc. Natl. Acad. Sci. USA 97: 14488-14493 [Abstract] [Full Text]  
  • Boerboom, D., Pilon, N., Behdjani, R., Silversides, D. W., Sirois, J. (2000). Expression and Regulation of Transcripts Encoding Two Members of the NR5A Nuclear Receptor Subfamily of Orphan Nuclear Receptors, Steroidogenic Factor-1 and NR5A2, in Equine Ovarian Cells during the Ovulatory Process. Endocrinology 141: 4647-4656 [Abstract] [Full Text]  
  • BURKE, L. J., BANIAHMAD, A. (2000). Co-repressors 2000. FASEB J. 14: 1876-1888 [Abstract] [Full Text]  
  • Wen, Y.-D., Perissi, V., Staszewski, L. M., Yang, W.-M., Krones, A., Glass, C. K., Rosenfeld, M. G., Seto, E. (2000). The histone deacetylase-3 complex contains nuclear receptor corepressors. Proc. Natl. Acad. Sci. USA 97: 7202-7207 [Abstract] [Full Text]  
  • Resnick, E. M., Schreihofer, D. A., Periasamy, A., Shupnik, M. A. (2000). Truncated Estrogen Receptor Product-1 Suppresses Estrogen Receptor Transactivation by Dimerization with Estrogen Receptors alpha and beta. J. Biol. Chem. 275: 7158-7166 [Abstract] [Full Text]  
  • Altincicek, B., Tenbaum, S. P., Dressel, U., Thormeyer, D., Renkawitz, R., Baniahmad, A. (2000). Interaction of the Corepressor Alien with DAX-1 Is Abrogated by Mutations of DAX-1 Involved in Adrenal Hypoplasia Congenita. J. Biol. Chem. 275: 7662-7667 [Abstract] [Full Text]  

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