Previous Article | Next Article 
Molecular and Cellular Biology, November 1998, p. 6653-6665, Vol. 18, No. 11
Centre de Recherche de Biochime
Macromoléculaire, CNRS UPR1142, 34293 Montpellier cedex 5, France,1 and
Institute of Human
Genetics, University of Freiburg, D-79106 Freiburg im Breisgau,
Germany2
Received 28 May 1998/Returned for modification 7 July 1998/Accepted 20 July 1998
For proper male sexual differentiation, anti-Müllerian
hormone (AMH) must be tightly regulated during embryonic development to
promote regression of the Müllerian duct. However, the molecular mechanisms specifying the onset of AMH in male mammals are not yet
clearly defined. A DNA-binding element for the steroidogenic factor 1 (SF-1), a member of the orphan nuclear receptor family, located in the
AMH proximal promoter has recently been characterized and demonstrated
as being essential for AMH gene activation. However, the requirement
for a specific promoter environment for SF-1 activation as well as the
presence of conserved cis DNA-binding elements in the AMH
promoter suggest that SF-1 is a member of a combinatorial protein-protein and protein-DNA complex. In this study, we demonstrate that the canonical SOX-binding site within the human AMH proximal promoter can bind the transcription factor SOX9, a Sertoli cell factor
closely associated with Sertoli cell differentiation and AMH
expression. Transfection studies with COS-7 cells revealed that SOX9
can cooperate with SF-1 in this activation process. In vitro and in
vivo protein-binding studies indicate that SOX9 and SF-1 interact
directly via the SOX9 DNA-binding domain and the SF-1 C-terminal
region, respectively. We propose that the two transcription factors
SOX9 and SF-1 could both be involved in the expression of the AMH gene,
in part as a result of their respective binding to the AMH promoter and
in part because of their ability to interact with each other. Our work
thus identifies SOX9 as an interaction partner of SF-1 that could be
involved in the Sertoli cell-specific expression of AMH during
embryogenesis.
In mammals, male sex determination
starts by activation of the testis-determining factor gene,
SRY, within cells of the supporting cell precursor lineage
(15, 44). When produced, SRY protein will then trigger
differentiation of these embryonic cells into Sertoli cells. After
differentiation, the Sertoli cells will export the male determining
signal via the production of a member of the transforming growth factor
An important initial finding came from deletion analysis of the AMH
promoter region that led to the identification of a 180-bp segment
required for correct AMH expression in primary Sertoli cells
(43). Characterization and analysis of this region in humans, bovines, mice, and rats (43) indicate that it
contains at least two highly conserved sequence elements plus a
characteristic TATA box (see Fig. 1A). The proximal element that
includes a single estrogen receptor half-site, AGGTCA, is
known to interact with a protein designated steroidogenic factor 1 (SF-1), the mammalian homologue of the Drosophila orphan
nuclear receptor fushi tarazu factor 1 (FTZ-F1), a factor
that regulates expression of the fushi tarazu homeobox gene
during early development (23, 28, 33). The functional
importance of this conserved SF-1-binding site was supported by several
observations such as its high conservation among species, its binding
in vitro to purified SF-1 protein, a coincident expression profile
between SF-1 and AMH, and the ability of a DNA fragment containing this
site to drive the transcription of a reporter gene in a Sertoli
cell-specific manner as demonstrated in transgenic animals (13,
43). However, the SF-1 binding site failed to activate gene
expression from the AMH promoter in heterologous cells such as HeLa
cells (43). This activation was shown to require removal of
the ligand-binding domain of the SF-1 protein, suggesting that a
Sertoli cell-specific ligand or cofactor must be necessary for SF-1 to
fulfil its transcriptional activity.
While the existence of a SOX-binding element in the 180-bp minimal
promoter led many investigators to postulate the testis-determining factor SRY as a candidate to control AMH expression, rather
contradictory results and hypotheses have been produced so far
(18, 20, 42, 43, 50). Among the additional sex-determining
genes described in recent years that were shown to be expressed
concomitant with or shortly after SRY, the
SRY-related gene SOX9 is the most attractive
candidate to contribute to this control. SOX9 was initially identified by positional cloning as associated with the skeletal malformation syndrome campomelic dysplasia, in which two-thirds of XY
individuals show sex reversal (10, 47). The recent detailed analysis of mouse Sox9 expression during gonadal development
coincident with Sertoli cell differentiation and its upregulation
preceding the onset of AMH expression in mice and chickens are the
first arguments to support such a hypothesis (3, 26, 32).
Furthermore, unlike human SRY, SOX9 was shown to act as a
transcriptional activator during chondrocyte differentiation (2,
30) and to display a high level of protein conservation across
vertebrate evolution.
We now address the possibility that the conserved SOX-binding sequence
present within the 180-bp proximal promoter region of the human AMH
gene is a binding site for SOX9. We demonstrate that SOX9 interacts
with this sequence and increases the expression of an AMH
promoter/reporter gene construct. We provide evidence for direct
protein-protein interaction between SOX9 and SF-1 and for additive
activation of the human AMH promoter by these proteins. We also
demonstrate that potentiation of SF-1 activity by SOX9 requires both
the SOX9 transactivation domain and the SF-1 ligand-binding domain. We
conclude that SOX9 and SF-1 have to function in a cooperative manner
for the proper control of AMH gene expression during early male gonadal
development.
Cloning and plasmid constructions.
Human SF-1 cDNA was
obtained by screening a
0270-7306/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.
Direct Interaction of SRY-Related Protein SOX9 and
Steroidogenic Factor 1 Regulates Transcription of the Human
Anti-Müllerian Hormone Gene
ABSTRACT
Top
Abstract
Introduction
Materials & Methods
Results
Discussion
References
INTRODUCTION
Top
Abstract
Introduction
Materials & Methods
Results
Discussion
References
family, the anti-Müllerian hormone (AMH), also known as
Müllerian inhibitory substance (for a review, see reference
25). AMH production promotes the regression of Müllerian ducts, the anlagen of the female reproductive organs, and thus appears to be critical for establishing the male phenotype. Molecular studies have been performed by several groups to reconstruct the pathway of primary male sex determination initiated by SRY and
terminated by AMH secretion by Sertoli cells. Since AMH expression will
follow Sertoli cell determination initiated by SRY, one attractive hypothesis was the direct control of AMH expression via the
SRY gene product, a high-mobility group (HMG) box containing
transcription factor (19-21). However, the time lag between
SRY expression and AMH expression (18)
and the absence of any transactivation domain in the human SRY protein
(8, 9) have led many investigators to refute this hypothesis
(13, 14, 43). It has been suggested that other genes in the
cascade located downstream of SRY have to fulfil this role. The recent
description of additional transcription factors involved in the
sex-determining pathway and the compilation of conserved DNA-binding
sites located in the AMH promoter sequences from diverse mammalian
species have opened new tracks for investigating the control of AMH
expression during male embryogenesis.
MATERIALS AND METHODS
Top
Abstract
Introduction
Materials & Methods
Results
Discussion
References
ZAPII human embryonic cDNA library
(24) with a human SF-1-derived genomic 153-bp fragment. This
fragment was the result of the reverse transcription-PCR amplification
of human embryonic total RNA with the help of primers (forward,
5'-GGTGTCCGGCTACCACTACGG-3'; reverse,
5'-CCAACGCCGACAAGGGACAGC-3') designed from the human
SF-1 partial genomic sequence deposited in GenBank under
accession no. U32592. The amplification product was next sequenced to
verify its identity to the SF-1 sequence. The SF-1-derived probe was
labeled with [
-32P]dCTP by random priming (Megaprime;
Amersham) and then used for the screening. SF-1 cDNA-containing
phagemids were excised in vivo from the ZAPII vector by coinfection
of Escherichia coli XL1-blue cells with VCSM13 helper phage
(Stratagene) as specified by the supplier and then fully sequenced.
This sequence is deposited in GenBank (accession no. U32591). Human
SOX9 cDNA was cloned in pcDNA3 vector (Invitrogen).
(i) Yeast expression plasmids. Full-length or partial SF-1 open reading frames, including the sequences encoding amino acids 1 to 461, 1 to 226, and 223 to 461, were obtained by PCR amplification with the appropriate oligonucleotides containing upstream BamHI sites and downstream SalI sites. PCR products were next cloned into pUC18 (SureClone ligation kit; Pharmacia) and checked by sequencing. The different fragments were then subcloned into the BamHI and SalI sites of pGBT11 as a fusion with the GAL4 DNA-binding domain. pGADGH-SOX9 constructs were obtained by inserting the human SOX9 open reading frame or the SOX9 fragment, spanning positions 1 to 304, from the pcDNA3 construct into the BamHI site of pGADGH (Clontech) as a fusion with the GAL4 activating-domain DNA sequence. Again, the authenticity of the constructs and ligation junctions was checked by sequencing.
(ii) Bacterium expression vectors. Both SF-1 and SOX9 proteins were bacterially expressed as glutathione S-transferase (GST) fusion proteins after PCR amplification of the corresponding cDNA and cloning into the BamHI and EcoRI sites of the pGEX-4T3 expression vector (Pharmacia). The authenticity of the constructs was checked by sequencing.
(iii) Mammalian reporter and expression plasmids.
To
construct the reporter gene plasmid, pEMBL8-AMH (16) was
used as a template to amplify AMH promoter DNA from positions +10 to
154 by PCR. The two primers contain the recognition site for either
SalI or SacI. After
SalI-SacI digestion, one copy of the 164-bp PCR
product was inserted in the SalI-SacI sites of the pEMBL-CAT vector (Stratagene) upstream of the chloramphenicol acetyltransferase (CAT) coding sequence. This construct is referred to
as p154CAT. The same strategy was used to produce the p123CAT construct. Mutagenesis of the SOX-binding site to produce p154MUTSOXCAT was performed by using the QuickChange site-directed mutagenesis kit
(Stratagene) with the help of the SOX-MUT oligonucleotide (see below).
Full-length SF-1 cDNA was cloned as a BamHI-EcoRI fragment into the pcDNA3 vector, which directs transcription from the
cytomegalovirus promoter. pcDNA-SOX9 and pcDNA-SOX9 1-304 were
described previously (46).
(iv) Plasmid constructs for in vitro translation. pcDNA-SOX9 HMG was described previously (31). The SOX9 1-118 deletion mutant was obtained by BssHII digestion of pcDNA-SOX9. For the other two deletions (SOX9 1-208 and SOX9 1-95), pcDNA-SOX9 was digested with BamHI and the released insert was cloned in pBluescript vector. This construct was finally digested with either PstI or HincII and ligated to create SOX9 1-208 and SOX9 1-95 mutants, respectively.
Synthesis of proteins in vitro and preparation of nuclear extracts. SOX9 protein, SOX9 mutants, and SF-1 protein were synthesized by in vitro transcription-translation with the expression vectors described above and with the TNT system (Promega). Nuclear extracts from NT2/D1 cells were prepared as described previously (41).
Production and purification of bacterially expressed pGEX-SOX9
and pGEX-SF-1 fusion proteins.
After being cloned in the pGEX-4T3
expression vector, the two recombinant proteins were produced in
bacterial strain BL21(DE3) after induction with 0.5 mM
isopropyl--D-thiogalactopyranoside (IPTG). After 2 h of induction at 30°C, cells were collected by centrifugation,
resuspended in lysis buffer (150 mM NaCl, 1 mM dithiothreitol [DTT],
5 mM EDTA, 25% sucrose, 50 mM Tris [pH 7.5]) supplemented with
bovine DNase I (Pharmacia) and 0.5 mM Pefabloc-SC-AEBSF (Interchim),
and sonicated for 5 min at 4°C. Bacterial debris were removed by
centrifugation at 25,000 rpm for 20 min. Lysate was loaded onto
glutathione-Sepharose beads and washed three times with buffer I (5 mM
EDTA, 250 mM NaCl, 50 mM Tris [pH 7.6]) and three times with buffer
II (5 mM EDTA, 120 mM NaCl, 50 mM Tris [pH 7.6]). The purified
proteins were checked by sodium dodecyl sulfate-polyacrylamide gel
electrophoresis (SDS-PAGE) analysis and used directly for in vitro
binding assay studies or eluted from the matrix with buffer II plus 10 mM reduced glutathione after a 30-min incubation at 4°C.
Production, purification, and characterization of SF-1 and SOX9
antibodies.
Polyclonal human SF-1-specific rabbit serum and rat
serum were raised against the GST fusion protein containing amino acids 121 to 232 from the human SF-1 protein. The fusion protein was overexpressed in BL21 (DE3) cells and purified on a
glutathione-Sepharose column as described above. Male New Zealand White
rabbits and rats were injected with purified protein mixed with
complete Freund's adjuvant (Sigma) and bled 10 days after each
injection. For purification, only the rabbit polyclonal antiserum was
loaded onto GST beads overnight and affinity purified by blotting
overnight onto the SF-1 peptide coupled to an Immobilon membrane
(Millipore). After saturation with polyvinylpyrrolidone (Sigma), SF-1
antibody was eluted with 0.2 M glycine (pH 2.5) and dialyzed against 1 M Tris base (pH 7.5). The antibody was finally aliquoted and stored at 80°C.
DNA-binding assays.
Protein binding to AMH promoter DNA
probes was assessed by the electrophoretic mobility shift assay (EMSA).
For these experiments, double-stranded oligonucleotides were labeled by
a fill-in reaction in the presence of [-32P]dCTP and
DNA polymerase I Klenow fragment for 1 h at 37°C. The labeled
probe was purified on a 5% nondenaturing acrylamide gel and eluted
overnight in 0.8 M ammonium acetate-5 mM EDTA-0.1% SDS buffer at
50°C. For the binding reaction, 5 to 10 µg of nuclear extract or
recombinant protein was mixed with 32P-labeled probe
(10,000 cpm) and 2 µg of poly(dI-dC) for SF-1 protein or 2 µg of
poly(dG-dC) for SOX9 protein in a 20-µl final volume of binding
buffer (20 mM HEPES [pH 7.9], 20% glycerol, 0.1 M KCl, 0.2 mM EDTA,
0.5 mM phenylmethylsulfonyl fluoride, 0.5 mM DTT). For competition or
supershift experiments, unlabeled competitor oligonucleotide or
antibody solution was incubated for 15 min at room temperature before
the probe was added. The DNA-protein complexes were resolved by
electrophoresis on a 5% polyacrylamide gel in Tris-borate-EDTA buffer
at 4°C and visualized by autoradiography after being fixed with 10%
methanol-10% acetic acid and dried.
Cell culture and transfection assays.
The human NT2/D1 cells
(N-Tera 2, clone D1, a human pluripotent embryonic carcinoma cell line
[ATCC CRL 1973]) were obtained from the American Type Culture
Collection (Biovaley, France). NT2/D1 and COS-7 cells were cultured in
Dulbecco's modified Eagle's medium (Imperial Laboratories, Flobio,
France) containing 10% (vol/vol) fetal calf serum (Life Technologies),
penicillin/streptomycin, and 2 mM glutamine with a partial pressure of
CO2 of 5% at 37°C in humidified air. Plasmids used for
transfections were purified with the maxiprep reagent system (Qiagen).
COS-7 or NT2/D1 cells at 60% to 80% confluence were washed twice with
serum-free medium before undergoing cotransfection with 1 µg of
reporter plasmid, 200 ng of pCMV--galactosidase plasmid
(Stratagene) used as an internal control for transfection efficiency,
and different SF-1 and SOX9 expression plasmids with 7 µl of
Lipofectamine (Life Technologies) in 200 µl of serum-free medium.
After a 6-h incubation, the medium was replaced with 2 ml of medium
supplemented with 10% serum and the cells were harvested after 48 h of culture. CAT assays were performed on cell extracts with
[3H]acetyl coenzyme A (200 mCi/mmol; Amersham, Little
Chalfont, United Kingdom) by a nonchromatographic method as described
by Nielsen et al. (37). Promoter activities were expressed
as CAT activity units per -galactosidase unit, and each value
represents the mean of the results from four separate wells. Error bars
represent the standard errors.
Protein interaction assays. For protein association experiments on glutathione-Sepharose beads, GST-SF-1 fusion proteins were overexpressed in BL21 bacteria and purified as described above. Immobilized SF-1 proteins were then incubated with the different 35S-labeled SOX9-derived proteins obtained by in vitro translation and the diverse SOX9 constructs in pcDNA3 and pBluescript vectors. Incubations were carried out in 300 µl of TBST buffer (10 mM Tris-HCl [pH 7.5], 130 mM NaCl, 0.5% Tween 20)-0.2% bovine serum albumin [BSA]-ethidium bromide (50 µg/ml) at room temperature for 30 min. The Sepharose beads were washed three times with 1 ml of TBST buffer. Bound proteins were eluted by the addition of 5× Laemmli buffer, boiled, and visualized after SDS-PAGE analysis and autoradiography.
Yeast two-hybrid interaction assays.
After the different
pGADGH-SOX9 constructs (containing the leucine-selective marker) were
obtained, each was transformed into the Mat Y187 yeast
strain by standard procedures. On the other hand, the Mat
Hf7c yeast strain was transformed with the different pGBT11-SF-1
constructs (containing the tryptophan-selective marker). These two
yeast strains both harbor HIS3 and -galactosidase
reporter genes under the control of GAL4-binding sites. Diploids were
obtained by mating and were selected on DO-W-L medium without
tryptophan and leucine, as reported previously (11).
Interaction assays were done for three independent transformants.
Histidine assays were conducted on Y187-H7fc diploid strains expressing
the designated constructs on DO-W-L-H medium without tryptophan,
leucine, and histidine. Quantitative -galactosidase assays were
conducted on the same diploids as the histidine assays, and mean values are given in -galactosidase units.
In vivo coimmunoprecipitation.
Detection of SF-1/SOX9
complexes was analyzed in vivo in the NT2/D1 cell line. After a
4-h labeling with 100 µCi of [35S]methionine
(Amersham; specific activity, >800 Ci/mmol), the cells were washed
with phosphate-buffered saline (PBS), collected, lysed for 30 min at
4°C in 1 ml of TBST or TLB (20 mM Tris-HCl [pH 7.6], 140 mM NaCl, 2 mM EDTA, 1% Triton X-100, 25 mM -glycerophosphate, 10% glycerol, 2 mM sodium pyrophosphate) buffer supplemented with Complete protease
inhibitor cocktail (Boehringer Mannheim) with or without 50 µg of
ethidium bromide per ml, and vortexed. All subsequent steps were
carried out on ice. Cell debris were removed, and the resulting lysate
was precleared with protein A-Sepharose beads (Pharmacia). Typically,
for each immunoprecipitation, 100 µl of cleared lysate was incubated
with anti-SF-1 antibody conjugated to protein A-Sepharose beads at
4°C in a total volume of 1 ml for 1 h with continuous rocking.
The beads were pelleted and washed five times in the lysis buffer, and
the resultant proteins were diluted in 5× Laemmli buffer and subjected
either to SDS-PAGE and autoradiography or to Western blot analysis with
the SOX9 antibody (diluted 1/400) revealed with an ECL kit (Amersham).
DNase I footprinting assay.
A double-stranded DNA fragment
corresponding to the 164-bp region from the AMH promoter was used for
DNase I footprinting analysis. Briefly, a pUC18-AMH 164-bp construct
was linearized by digestion with BamHI and labeled with
[-32P]dCTP. The 164-bp fragment was then released by
EcoRI. The labeled fragment was gel purified and eluted at
50°C in 0.8 M ammonium acetate-5 mM EDTA-0.1% SDS buffer. For each
footprinting reaction, 104 cpm of the probe was added to
various amounts of purified SF-1 and SOX9 proteins in 50 µl of
mixture containing 10 mM HEPES (pH 7.8), 40 mM KCl, 3 mM
MgCl2, 0.5 mM DTT, 5% glycerol, and 100 ng of poly(dI-dC)
for SF-1 or 100 ng of poly(dG-dC) for SOX9 and the mixture was
incubated at room temperature for 30 min. DNase I digestion was
performed by adding 1 U of enzyme diluted in the corresponding buffer
(Boehringer-Mannheim) and incubating the mixture for 1 min at 25°C.
The reaction was terminated by adding phenol-chloroform. The digested
fragments were recovered by ethanol precipitation, resuspended in 3 µl of stop buffer (0.1% xylene cyanol, 0.1% bromophenol blue, 10 mM
EDTA, 95% formamide), incubated for 2 min at 90°C, and resolved by
electrophoresis on a 6% polyacrylamide-urea gel. A DNA ladder was
made by dimethyl sulfide treatment for 5 min of 2 µl of labeled probe
and piperidine treatment at 90°C for 30 min. This ladder was used to
assign nucleotide positions in the gel.
Immunofluorescence staining of tissue culture cells. Cells were preincubated in PBS buffer-1% BSA for 30 min at 37°C and then probed with the appropriate antibody, anti-SF-1, anti-SOX9, or anti-AMH diluted 1/100 in PBS/BSA. The cells were washed in PBS, and the primary antibodies were visualized with biotinylated anti-rabbit antibodies (dilution, 1/200) and Texas red-conjugated streptavidin or with Fluorolink Cy2-conjugated anti-rat labeled goat antibodies (dilution, 1/200). In each case, incubations were performed for 30 min under the same conditions described for the primary antibodies. For colocalization analysis, incubation with both appropriate primary antibodies was performed in the same incubation buffer and the same antibody dilutions were used with the secondary antibodies. Cell nuclei were visualized with Hoechst 33286. The cells were washed again and mounted in FluorSave reagent (Calbiochem). Images were collected and processed on a Bio-Rad confocal microscope or on a Zeiss Axiophot.
| |
RESULTS |
|---|
|
Top
Abstract
Introduction
Materials & Methods
Results
Discussion
References
|
|---|
A consensus and functional SOX-binding site resides within the
proximal AMH promoter.
Previous in vitro studies of the AMH
promoter indicated that not more than 180 bp is required for proper AMH
expression in Sertoli cells (43). Diverse observations
including the presence of a conserved SF-1 site (Fig.
1A), mutation analysis, or transgenic experiments have revealed the requirement for this site in AMH gene
control (13, 43). However, several results have also pointed
out the necessity for a specific Sertoli-related cell environment for
activation, indicating that an uncharacterized Sertoli cell factor
contributes to this activation (43). We now confirm this
requirement by using the COS-7 cell line as the transfection control
(Fig. 1B). Pursuing progressive deletions of the minimal human AMH
promoter, we show that the nucleotide sequence between 154 and 123
of the AMH promoter is important for promoter function, as demonstrated
by transfection assays in the NT2/D1 cell line (Fig. 1B). The NT2/D1
cell line still constitutes one of the rare cell models positive for
SRY expression (4, 41). Inspection of the nucleotide
sequence confirmed previous analysis by revealing a 7-of-8-bp identity
(5'-CCTTGAG, referred to as the SOX site in this study) to a
known binding site that represents a potential site for SRY and other
related members of the HMG-box class DNA-binding factors such as SOX, TCF-1, or LEF-1 protein (12, 22, 30, 36). Finally, using direct mutagenesis, we also show the necessity for the SOX site in such
an additive activation (Fig. 1B).
|
Purified human SOX9 and SF-1 proteins bind to the AMH promoter in
vitro.
In vitro DNase I footprint analysis showed that purified
GST-SOX9 protein could protect the 154 to 135 sequence from the AMH
proximal promoter, the sequence spanning the SOX site (Fig. 2A). As a control, by using the same
164-bp labeled oligonucleotide, the same experimental conditions, and
the purified GST-SF-1 fusion protein, we also confirmed the protection
of the 97 to 82 DNA sequence that includes the SF-1 binding site
previously described (43) (Fig. 2B). A labeled 28-bp
oligonucleotide probe including the SOX-binding site was next checked
in EMSA experiments with GST-SOX9 protein. As shown in Fig. 2C, a
single DNA protein complex was evident. Complex formation was blocked
by using the same unlabeled oligonucleotide as the competitor. When a
double mutation known to abolish SOX protein binding was introduced
into the oligonucleotide (SOX-MUT), the competition was abolished.
These results indicate that SOX9 binds directly to the DNA in the
region of the AMH promoter. On the other hand, similar EMSA experiments
with purified GST-SF-1 and the corresponding SF-1
oligonucleotide-binding site designed from the AMH proximal promoter
confirm SF-1 protein interaction (Fig. 2D).
|
Transactivation of AMH expression by SOX9 and by SF-1 in COS-7 cells. The functional relevance of SOX9 factor binding to the SOX site located in the AMH proximal promoter and the possibility of its cooperation with the SF-1 nuclear receptor in activating the AMH promoter were examined. Cotransfections of COS-7 cells with a reporter plasmid which contains the 164-bp AMH DNA fragment adjacent to the CAT gene and termed p154CAT (see Materials and Methods), together with increasing SF-1 expression plasmid concentrations but with a fixed SOX9 plasmid concentration, were performed (Fig. 3A). The use of 10 ng of pcDNA3-SOX9 along with 200 ng of pcDNA3-SF-1 provides the best activation signal (more than fivefold compared with the empty vector) and was used in the following experiments (Fig. 3A). As a control, under these experimental conditions the specificity of this activation was then tested with a truncated form of SOX9 (deletion of amino acids 305 to 509), a form described as being able to bind the DNA target but unable to activate transcription (30). As shown in Fig. 3B, the SOX9 C-terminal transactivation domain was required for this activation. Similarly, no activation was observed when the effector plasmid expressing SRY, a previous candidate factor for contributing to AMH gene activation, was used (Fig. 3B). While this result confirmed recent data obtained with an artificial promoter (8), it also showed that SRY cannot substitute for SOX9 in the AMH-positive control. If the SOX9 activation domain appears necessary in the activation effect, its DNA-binding capacity is also required, as demonstrated when using a mutated version of the AMH proximal promoter (Fig. 3C).
|
SOX9 and SF-1 interact directly with each other as shown by two-hybrid analysis and by in vitro assays. The results just presented and the observation of rather closely spaced SOX- and SF-1-binding sites within the proximal AMH promoter region (Fig. 1B), along with the ability of SOX proteins to bend DNA (12), raised the possibility that the two factors interact with each other, as recently reported for another system involving Sox2 and Oct-3 (1, 52).
To obtain evidence for such an interaction, we first used the yeast two-hybrid assay and a mating protocol (11). For this, the region encompassing the SF-1 open reading frame was fused to the GAL4 DNA binding domain in the pGBT11 vector and mated with the SOX9 open reading frame fused to the GAL4 activation domain in the pGADGH vector. The yeast diploid transformants were able to grow on selective medium lacking histidine and were also-galactosidase positive (Fig.
4B and C). This result indicates an
interaction between the two proteins. We subsequently tested potential
interactions between full-length SOX9 and different regions of SF-1, as
depicted in Fig. 4A. We found that the carboxy-terminal region of SF-1
(amino acids 225 to 461) was sufficient for this interaction.
Correspondingly, SOX9 did not interact with the N-terminal portion of
SF-1 (amino acids 1 to 226). It is worth noting that identical results
were obtained when using only the N-terminal part of SOX9 (amino acids
1 to 304) (Fig. 4B). To quantify the strength of the interactions
between the different constructs of SF-1 and SOX9 proteins,
-galactosidase activity was measured from the transformant lysates
(Fig. 4C). These results confirm that the strongest interaction occurs
between the N-terminal region of SOX9 and the C-terminal region of
SF-1, including its ligand-binding domain. Although these results
demonstrate that SOX9 and SF-1 interact, we cannot exclude the
contribution of a posttranslational modification or the involvement of
a third partner present in the yeast strain and contributing to the
interaction.
|
|
SOX9 and SF-1 interact in vivo. We then examined whether SOX9 and SF-1 form a protein complex in vivo. The ability of SOX9 to interact directly with SF-1 independently of the SOX9 DNA-binding activity was first tested by EMSA in the presence of NT2/D1 cell nuclear extracts and the labeled SF-1 binding-site oligonucleotide whose sequence was derived from the AMH proximal promoter. Nuclear extracts of NT2/D1 cells gave rise to a major protein-DNA complex (Fig. 6, lane 1), a DNA binding that was inhibited by an excess of unlabeled SF-1 oligonucleotide (lane 2) but not by a mutated SF-1 oligonucleotide (lane 10). Evidence for the identity of the retarded major band was obtained in supershift experiments. Preincubation of nuclear extracts in the presence of SOX9 or SF-1 antisera produced a supershift band in both cases (lanes 4 and 6). By contrast, addition of SRY antiserum did not modify the migration of the complex (lane 7), in agreement with previous results (43).
|
|
SOX9 and SF-1 colocalize in nuclei of NT2/D1 cells. We next analyzed the subcellular localization of the SOX9 and SF-1 proteins by performing indirect-immunofluorescence labeling experiments. The use of purified rabbit anti-SF-1 and rat anti-SOX9 antibodies allowed double-labeling experiments on the NT2/D1 cells. Both antibodies revealed a nuclear, punctuate localization for the two proteins, an expression throughout the cell culture (Fig. 8A and B). Cytoplasmic labeling of the AMH protein was observed in the same cells (Fig. 8C). We further analyzed the possibility of subcellular colocalization of SOX9 with SF-1. A good, if incomplete, colocalization between the two proteins was observed after examination by confocal laser-scanning microscopy (Fig. 8F). These results strengthen the notion that SOX9 and SF-1 interact in vivo, possibly as part of a multimeric protein complex.
|
| |
DISCUSSION |
|---|
|
Top
Abstract
Introduction
Materials & Methods
Results
Discussion
References
|
|---|
In the developing gonad, when the sex-determining factor Sry is present, the supporting cell precursors differentiate along the Sertoli cell pathway. The first known product of the embryonic Sertoli cell is AMH. Initiation, maintenance, and down-regulation of AMH expression by Sertoli cells still remain a matter of controversy. If AMH has to be tightly regulated during gestation in order to avoid Müllerian duct persistence (17), it also requires a sexually dimorphic regulation because of its necessary absence in females during embryogenesis.
The aim of the work described here was to investigate the functional
relationship between cis-acting conserved elements located in the AMH proximal promoter in order to contribute to our
understanding of the transcription factors controlling AMH promoter
activity and leading to its spatiotemporal regulation during
embryogenesis. Sequence analysis showed conserved binding sites for
different transcription factors within the proximal and minimal 180-bp
AMH promoter (13, 43). In this study, the functional
importance of the nuclear receptor SF-1 binding site located in the AMH
promoter and previously revealed by studies with primary Sertoli cells (43) was confirmed. However, these results also shed light
on the requirement for an appropriate developmental context or cell environment for SF-1 activation of AMH, suggesting that a ligand or a
cofactor (or both) for SF-1 is needed (43). As mentioned in
this and other reports (18, 43, 50), apart from the now well-described nuclear receptor SF-1 site, a nearly perfect match with
the consensus SOX-binding site appears conserved in the promoter of
distantly related species. Performing progressive 5' deletions of the
AMH proximal promoter revealed that a deletion of the sequence from
154 to 123 which removes this SOX-binding site caused a strong
decrease in the basal activity of the AMH proximal promoter after
transfection into the "Sertoli-like" human NT2/D1 cell line. This
result was confirmed after mutation of the candidate SOX site.
Functional in vitro analysis of this site by either DNase I
footprinting or in vitro DNA-binding experiments showed that this
putative site was able to specifically bind bacterially expressed SOX9
protein. The choice of this particular SOX protein was dictated by
several arguments, including its conservation among vertebrates and its
high level of expression closely correlating with Sertoli cell
differentiation and subsequent AMH production during testicular development in mice and chickens (32) as well as in humans
(unpublished data). Furthermore, the sex reversal phenotype observed in
campomelic dysplasia patients carrying an SOX9 mutant is consistent
with failure of Sertoli cell differentiation and subsequent absence of
AMH production (31). Finally, of the two SOX proteins, SRY and SOX9, that have been identified as being expressed in the male
sex-determining pathway, only SOX9 has been shown to act as a potent
transcriptional activator in humans via a transactivation domain mapped
to its C terminus (30, 46). As suggested by Ambrosetti et
al. (1), the requirement for DNA-sequence-specific HMG
domain transcription factors such as Sox2 (1, 52), Sox5 (7), and TCF-1 (48) in a particular promoter
environment suggests that they are unable to act autonomously but,
instead, must collaborate with other transcription factors. Under our
experimental conditions with the COS-7 cell line,
transient-cotransfection experiments with the SOX9 expression plasmid
showed that SOX9 can activate AMH promoter activity to an extent
similar to that for SF-1 (data not shown) even when using a single copy
of the AMH proximal promoter. Thus, the kidney-derived cell line COS-7 appears to provide a convenient AMH promoter environment to test SF-1
or SOX9 function. Furthermore, these transactivation experiments also
confirmed the strict requirement for the C-terminal domain of SOX9,
implying that AMH promoter activation by SOX9 is not simply the result
of the bending capacity of its HMG domain. In contrast, despite a 71%
amino acid similarity to SOX9 in the HMG domain (10),
the SOX transcription factor family founder SRY was ineffective in this
activation. Because of the ability of both SOX9 and SF-1 to cooperate
in AMH proximal promoter activation, it was tempting to investigate if
this cooperation involved a protein-protein interaction event. Five
different and complementary approaches, namely, two-hybrid analysis,
GST pulldown assays, EMSA, coimmunoprecipitation experiments, and
immunolocalization studies, all indicate that SOX9 and SF-1 can
interact with each other and may be part of a multimeric protein
complex. We then performed a preliminary mapping of the amino acid
sequence responsible for this interaction. Dissection of the
transcription factor SOX9 and subsequent coprecipitation assays with
GST-SF-1 showed that the conserved N-terminal region of the SOX9 DNA
binding domain is required for its interaction with SF-1. This region
of SOX9 contains several amino acid sequence stretches that are
conserved between different members of the SOX family and might provide the basis for interaction of SF-1 with other members of the family. Interestingly, another Sox gene product, Sox2, was recently shown to
cooperate with the octamer-binding protein Oct-3 in order to synergistically activate the fibroblast growth factor 4 enhancer (1, 52). This activation was dependent both on the
protein-protein interaction involving the HMG domain of Sox2 and on the
presence of DNA-binding sites for both Sox2 and Oct-3 transcription
factors (1). A similar result was obtained with the
SOX9/SF-1 couple. We propose that SOX9 could, by its DNA-bending
activity (30), induce a local architectural modification of
the DNA target, allowing the formation of a transcription complex
including SF-1. The SOX9-SF-1 interaction could then stabilize the
resulting protein-DNA complex. This hypothesis is now under
investigation. On the other hand, the region of SF-1 that was
characterized in the two-hybrid experiment described herein as being
required for the interaction starts after the proline-rich region of
the receptor and extends into the ligand-binding and dimerization
domain including both activation domains of SF-1 (6). This
region has been previously reported as supporting the cell specificity
of SF-1 activation of AMH reporter constructs (43) as well
as SF-1 activation by oxysterol (29); it also permits
interaction with the nuclear receptor DAX-1 (5). However,
despite the possible overlap between the two regions of SF-1 involved
in oxysterol activation and SOX9 binding, the SF-1-SOX9
interaction was not modified in the presence of increasing 25-hydroxycholesterol concentrations (data not shown). The
ability of this SF-1 sequence region to interact with the coactivator SRC-1 was also recently shown (6). This interaction would
allow bridging of the transcription factors such as SOX9 and SF-1 that govern the tissue-specific expression of AMH and the basal
transcriptional machinery.
The absence of AMH expression before 12.5 days postcoitum in the mouse embryo despite coincident expression of both SF-1 and Sox9 could be questionable (34). This apparent contradiction with our data could be the result either of the absence of an elusive partner or of a missing positive regulation affecting one of the two transcription factors before this developmental stage. Another possibility is a regulation of the subcellular localization of one or more transcription factors contributing to the control of the onset of AMH expression. In this respect, mouse Sox9 expression studies have revealed a predominant cytoplasmic expression in cells of the genital ridge prior to 11.5 days postcoitum, i.e., prior to the sex determination event. At later stages, Sox9 appeared fully nuclear in male embryonic gonads (32). As recently suggested (45), the change in Sox9 localization could be achieved via its interaction with putative protein partner(s) that would mask the Sox9 nuclear localization signal(s). As an alternative, or in parallel, the mechanism used in this cytoplasm-nucleus trafficking could result from the existence of a leucine motif in the HMG domain of Sox9 (10, 47), a motif conserved across SOX9 evolution but also specific for this particular SOX protein (39). This kind of motif has been reported in many cases to act as a nuclear export signal (49). This hypothesis will not require further investigations.
Other gene products, namely, the Wilms' tumor WT1 and the nuclear
receptor DAX-1, are also implicated in mammalian sexual development.
Recently, WT1 in its KTS isoform has been shown to interact and
synergize with SF-1 (35). In the same report, DAX-1 was also
shown to antagonize this synergy, even when present at low
concentration. The inhibitory action of DAX-1 was accompanied by its
ability to interact directly with the SF-1 ligand-binding domain.
Interestingly, SOX9- and DAX-1-binding sites could overlap with respect
to their interaction with SF-1. Furthermore, in the same report, both
WT1 and DAX-1 expression levels appear rather similar between male and
female mouse embryos 13.5 days postcoitum, i.e., at a time when AMH
expression is on (35). Therefore, the only difference
between the sexes remains in the expression of the putative
transcription factors SRY and SOX9. If, as mentioned above, SRY does
not provide an attractive candidate, the high levels of SOX9 observed
specifically in male embryos (32) could compete with DAX-1
for its binding to SF-1 and thus could permit WT1 action. This
hypothesis would justify the rather low level of AMH stimulation
observed in the present data when using SF-1 and SOX9 only.
It is well established that transcriptional regulation of a given gene is the result of combinatorial interactions between multiple proteins forming a higher-order complex based on protein-protein and protein-DNA interactions. We now suggest that AMH gene regulation is no exception to this general rule and that both SF-1 and SOX9 are members of the complex regulating the onset of AMH expression during embryogenesis beyond WT1 and DAX-1. This statement will now require introduction of these four transcription factors in the same assay as well as in vivo characterization. The two proteins SOX9 and SF-1 appear to bind independently to separate DNA sites and could, especially because of the DNA-bending capability of SOX9 (30), facilitate the functional interaction of other regulatory proteins, leading to the formation of the appropriate transcription complex triggering Sertoli-specific AMH expression. Interestingly, another well-conserved binding site in the AMH proximal promoter has homology to the in vitro canonical binding site for the GATA transcription factor family, WGATAR (Fig. 1A). Expression of GATA-1 or GATA-2 factors in Sertoli cells reinforces this hypothesis and makes GATA factors attractive candidates to contribute to the regulation of AMH expression (38, 51), a hypothesis that we are now attempting to test.
| |
ACKNOWLEDGMENTS |
|---|
We thank V. Laudet for the gift of the c-erbA construction plasmid, R. Rey for the rabbit anti-AMH antibody, J.-Y. Picard for pEMBL8-AMH plasmid, and A. Goldsborough and V. Laudet for comments on and corrections of the manuscript. We are grateful to Catherine Méjean for protein production and purification assistance and to Sandrine Faure for his constant support.
This investigation was supported by Biomed 2 grant BMH4-CT96-0790 from the European Economic Community.
| |
FOOTNOTES |
|---|
* Corresponding author. Present address: Human Molecular Genetics Group, Institut de Génétique Humaine, 141 rue de la cardonille, 34396 Montpellier cedex 5, France. Phone: (33) 499619955. Fax: (33) 499619901. E-mail: berta{at}igh.cnrs.fr.
| |
REFERENCES |
|---|
|
Top
Abstract
Introduction
Materials & Methods
Results
Discussion
References
|
|---|
| 1. | Ambrosetti, D.-C., C. Basilico, and L. Dailey. 1997. Synergistic activation of the fibroblast growth factor 4 enhancer by Sox2 and Oct-3 depends on protein-protein interactions facilitated by a specific spatial arrangement of factor binding sites. Mol. Cell. Biol. 17:6321-6329[Abstract]. |
| 2. | Bell, D. M., K. K. H. Leung, S. C. Wheatley, L. J. Ng, S. Zhou, K. W. Ling, M. H. Sham, P. Koopman, P. P. L. Tam, and K. S. E. Cheah. 1997. SOX9 directly regulates the type-II collagen gene. Nat. Genet. 16:174-178[Medline]. |
| 3. |
Carré-Eusèbe, D.,
N. Di Clemente,
R. Rey,
C. Pieau,
B. Vigier,
N. Josso, and J.-Y. Picard.
1996.
Cloning and expression of the chick anti-Müllerian hormone gene.
J. Biol. Chem.
271:4798-4804 |
| 4. |
Clépet, C.,
A. J. Schafer,
A. H. Sinclair,
M. S. Palmer,
R. Lovell-Badge, and P. N. Goodfellow.
1993.
The human SRY transcript.
Hum. Mol. Genet.
2:2007-2012 |
| 5. |
Crawford, P. A.,
C. Dorn,
Y. Sadovsky, and J. Milbrandt.
1998.
Nuclear receptor DAX-1 recruits nuclear receptor corepressor N-CoR to steroidogenic factor 1.
Mol. Cell. Biol.
18:2949-2956 |
| 6. |
Crawford, P. A.,
J. A. Polish,
G. Ganpule, and Y. Sadovski.
1997.
The activation function-2 hexamer is required, but not sufficient, for potentiation by SRC-1.
Mol. Endocrinol.
11:1626-1635 |
| 7. | Denny, P., S. Swift, F. Connor, and A. Ashworth. 1992. An SRY-related gene expressed during spermatogenesis in the mouse encodes a sequence-specific DNA-binding protein. EMBO J. 11:3705-3712[Medline]. |
| 8. |
Desclozeaux, M.,
F. Poulat,
P. de Santa Barbara,
J.-P. Capony,
P. Turowski,
P. Jay,
C. Mejean,
B. Moniot,
B. Boizet, and P. Berta.
1997.
Phosphorylation of an N-terminal motif enhances DNA-binding activity of the human SRY protein.
J. Biol. Chem.
273:7988-7995 |
| 9. |
Dubin, R. A., and H. Ostrer.
1994.
Sry is a transcriptional activator.
Mol. Endocrinol.
8:1182-1192 |
| 10. | Foster, J. W., M. A. Dominguez-Steglich, S. Guioli, C. Kwok, P. A. Weller, M. Stevanovic, J. Weissenbach, S. Mansour, I. D. Young, P. N. Goodfellow, J. D. Brook, and A. J. Schafer. 1994. Campomelic dysplasia and autosomal sex reversal caused by mutations in an SRY-related gene. Nature 372:525-530[Medline]. |
| 11. | Fromont-Racine, M., J.-C. Rain, and P. Legrain. 1997. Toward a functional analysis of the yeast genome through exhaustive two-hybrid screens. Nat. Genet. 16:277-282[Medline]. |
| 12. | Giese, K., J. Cox, and R. Grosschedl. 1992. The HMG domain of lymphoid enhancer factor-1 bends DNA and facilitates assembly of functional nucleoprotein structures. Cell 69:185-195[Medline]. |
| 13. | Giuili, G., W.-H. Shen, and H. A. Ingraham. 1997. The nuclear receptor SF-1 mediates sexually dimorphic expression of Müllerian inhibiting substance, in vivo. Development 124:1799-1807[Abstract]. |
| 14. | Greenfield, A., and P. Koopman. 1996. SRY and mammalian sex determination. Curr. Top. Dev. Biol. 34:1-23[Medline]. |
| 15. | Gubbay, J., J. Collignon, P. Koopman, B. Capel, A. Economou, A. Munsterberg, N. Vivian, P. Goodfellow, and R. Lovell-Badge. 1990. A gene mapping to the sex-determining region of the mouse Y chromosome is a member of a novel family of embryonically expressed genes. Nature 346:245-250[Medline]. |
| 16. |
Guerrier, D.,
L. Boussin,
S. Mader,
N. Josso,
A. Khan, and J.-Y. Picard.
1990.
Expression of the gene for anti-Müllerian hormone.
J. Reprod. Fertil.
88:695-706 |
| 17. |
Guerrier, D.,
D. Tran,
J. M. Vanderwinden,
S. Hideux,
L. Van Outryve,
L. Legeai,
M. Bouchard,
G. Van Vliet,
M. H. De Laet,
J. Y. Picard, and N. Josso.
1989.
The persistent Müllerian duct syndrome: a molecular approach.
J. Clin. Endocrinol. Metab.
68:46-52 |
| 18. | Hacker, A., B. Capel, P. Goodfellow, and R. Lovell-Badge. 1995. Expression of Sry, the mouse sex determining gene. Development 121:1603-1614[Abstract]. |
| 19. |
Haqq, C. M.,
C. Y. King,
P. K. Donahoe, and M. A. Weiss.
1993.
SRY recognizes conserved DNA sites in sex-specific promoters.
Proc. Natl. Acad. Sci. USA
90:1097-1101 |
| 20. |
Haqq, C. M.,
C.-Y. King,
E. Ukiyama,
S. Falsafi,
T. N. Haqq,
P. K. Donahoe, and M. A. Weiss.
1994.
Molecular basis of mammalian sexual determination: activation of Müllerian inhibiting substance gene expression by SRY.
Science
266:1494-1500 |
| 21. | Harley, V. R., and P. N. Goodfellow. 1994. The biochemical role of SRY in sex determination. Mol. Reprod. Dev. 39:184-193[Medline]. |
| 22. |
Harley, V. R.,
D. Jackson,
P. Hextall,
J. R. Hawkins,
G. D. Berkovitz,
S. Sockanathan,
R. Lovell-Badge, and P. N. Goodfellow.
1992.
DNA binding activity of recombinant SRY from normal males and XY females.
Science
255:453-456 |
| 23. |
Honda, S.,
K. Morohashi,
M. Nomura,
H. Takeya,
M. Kitajima, and T. Omura.
1993.
Ad4BP regulating steroidogenic P-450 gene is a member of steroid hormone receptor superfamily.
J. Biol. Chem.
268:7494-7502 |
| 24. | Jay, P., S. Diriong, S. Taviaux, N. Roeckel, M.-G. Mattéi, M. Audit, J.-L. Bergé-Lefranc, M. Fontes, and P. Berta. 1996. Isolation and regional mapping of cDNAs expressed during early human development. Genomics 39:104-108. |
| 25. | Josso, N., R. L. Cate, J.-Y. Picard, B. Vigier, N. Di Clemente, C. Wilson, S. Imbeaud, R. B. Pepinsky, D. Guerrier, L. Boussin, L. Legeai, and D. Carré-Eusèbe. 1993. Anti-Müllerian hormone the Jost factor. Recent Prog. Horm. Res. 8:379-418. |
| 26. | Kent, J., S. C. Wheatley, J. E. Andrews, A. H. Sinclair, and P. Koopman. 1996. A male-specific role for SOX9 in vertebrate sex determination. Development 122:2813-2822[Abstract]. |
| 27. |
Lai, J.-S., and W. Herr.
1992.
EtBr provides a simple tool for identifying genuine DNA-independent protein associations.
Proc. Natl. Acad. Sci. USA
89:6958-6962 |
| 28. |
Lala, D. S.,
D. A. Rice, and K. L. Parker.
1992.
Steroidogenic factor 1, a key regulator of steroidogenic enzyme expression, is the mouse homolog of fushi tarazu-factor 1.
Mol. Endocrinol.
6:1249-1258 |
| 29. |
Lala, D. S.,
P. M. Syka,
S. B. Lazarchik,
D. J. Mangelsdorf,
K. L. Parker, and R. A. Heyman.
1997.
Activation of orphan nuclear receptor steroidogenic factor 1 by oxysterols.
Proc. Natl. Acad. Sci. USA
94:4895-4900 |
| 30. |
Lefebvre, V.,
W. Huang,
V. R. Harley,
P. N. Goodfellow, and B. de Crombrugghe.
1997.
SOX9 is a potent activator of the chondrocyte-specific enhancer of the pro1(II) collagen gene.
Mol. Cell. Biol.
17:2336-2346[Abstract].
|
| 31. |
Meyer, J.,
P. Südbeck,
M. Held,
T. Wagner,
M. L. Schmitz,
F. D. Bricarelli,
E. Eggermont,
U. Friedrich,
O. A. Hass,
A. Kobelt,
J. G. Leroy,
L. van Maldergem,
E. Michel,
B. Mitulla,
R. A. Pfeiffer,
A. Schinzel,
H. Schmidt, and G. Scherer.
1997.
Mutational analysis of the SOX9 gene in campomelic dysplasia and autosomal sex reversal: lack of genotype/phenotype correlations.
Hum. Mol. Genet.
6:91-98 |
| 32. | Morais da Silva, S., A. Hacker, V. Harley, P. Goodfellow, A. Swain, and R. Lovell-Badge. 1996. Sox9 expression during gonadal development implies a conserved role for the gene in testis differentiation in mammals and birds. Nat. Genet. 14:62-68[Medline]. |
| 33. |
Morohashi, K. I.,
S. L. Honda,
Y. Inamata,
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 |
| 34. | Münsterberg, A., and R. Lovell-Badge. 1991. Expression of the mouse anti-Müllerian hormone gene suggests a role in both female and male sexual differentiation. Development 113:613-624[Abstract]. |
| 35. | Nachtigal, M. W., Y. Hirokawa, D. L. Enyeart-VanHouten, J. N. Flanagan, G. D. Hammer, and H. A. Ingraham. 1998. Wilms' tumor 1 and Dax-1 modulate the orphan nuclear receptor SF-1 in sex-specific gene expression. Cell 93:445-454[Medline]. |
| 36. | Nasrin, N., C. Buggs, X. F. Kong, J. Carnazza, M. Goebl, and M. Alexander-Bridges. 1991. DNA-binding properties of the product of the testis-determining gene related protein. Nature 354:317-320[Medline]. |
| 37. | Nielsen, D. A., T. C. Chang, and D. J. Shapiro. 1989. A highly sensitive, mixed-phase assay for chloramphenicol acetyl transferase activity in transfected cells. Anal. Biochem. 179:19-23[Medline]. |
| 38. |
Onodera, K.,
K. Yomogida,
N. Suwabe,
S. Takahashi,
Y. Muraosa,
N. Hayashi,
E. Ito,
L. Gu,
M. Rassoulzadegan,
J. D. Engel, and M. Yamamoto.
1997.
Conserved structure, regulatory elements, and transcriptional regulation from the GATA-1 gene testis promoter.
J. Biochem.
121:251-263 |
| 39. | Pevny, L. H., and R. Lovell-Badge. 1997. Sox genes find their feet. Curr. Opin. Genet. Dev. 7:338-344[Medline]. |
| 40. |
Poulat, F.,
P. de Santa Barbara,
M. Desclozeaux,
S. Soullier,
B. Moniot,
N. Bonneaud,
B. Boizet, and P. Berta.
1997.
The human testis determining factor SRY binds a nuclear factor containing PDZ protein interaction domains.
J. Biol. Chem.
272:7167-7172 |
| 41. | Poulat, F., F. Girard, M.-P. Chevron, C. Gozé, X. Rebillard, B. Calas, N. Lamb, and P. Berta. 1995. Nuclear localization of the testis determining gene product SRY. J. Cell Biol. 5:737-748. |
| 42. | Schafer, A. J., and P. N. Goodfellow. 1996. Sex determination in humans. Bioessays 18:955-963[Medline]. |
| 43. | Shen, W. H., C. C. D. Moore, Y. Ikeda, K. L. Parker, and H. A. Ingraham. 1994. Nuclear receptor steroidogenic factor 1 regulates the Müllerian inhibiting substance gene: a link to the sex determination cascade. Cell 77:651-661[Medline]. |
| 44. | Sinclair, A. H., P. Berta, M. S. Palmer, J. R. Hawkins, B. L. Griffiths, M. J. Smith, J. W. Foster, A. M. Frischauf, R. Lovell-Badge, and P. N. Goodfellow. 1990. A gene from the human sex-determining region encoding a protein with homology to a conserved DNA-binding motif. Nature 346:240-244[Medline]. |
| 45. |
Südbeck, P., and G. Scherer.
1997.
Two independent nuclear localization signals are present in the DNA-binding high mobility group domains of SRY and SOX9.
J. Biol. Chem.
272:27848-27852 |
| 46. | Südbeck, P., M. L. Schmitz, P. A. Baeuerle, and G. Scherer. 1996. Sex reversal by loss of the C-terminal transactivation domain of human SOX9. Nat. Genet. 13:230-232[Medline]. |
| 47. | Wagner, T., J. Wirth, J. Meyer, B. Zabel, M. Held, J. Zimmer, J. Pasantes, F. D. Bricarelli, J. Keutel, E. Hustert, U. Wolf, N. Tommerup, W. Schempp, and G. Scherer. 1994. Autosomal sex reversal and campomelic dysplasia are caused by mutations in and around the SRY related gene SOX9. Cell 79:1111-1120[Medline]. |
| 48. |
Waterman, M., and K. Jones.
1990.
Purification of TCF-1, a T-cell-specific transcription factor that activates the T-cell receptor gene enhancer in a context-dependent manner.
New Biol.
2:621-636[Medline].
|
| 49. | Wen, W., J. L. Meinkoth, R. Y. Tsien, and S. S. Taylor. 1995. Identification of a signal for rapid export of proteins from the nucleus. Cell 82:463-473[Medline]. |
| 50. | Werner, M. H., J. R. Huth, A. M. Gronenborg, and G. M. Clore. 1995. Molecular basis of human 46X,Y sex reversal revealed from the three dimensional solution structure of the human SRY-DNA complex. Cell 81:705-714[Medline]. |
| 51. | Yomogida, K., H. Ohtani, H. Harigae, E. Ito, Y. Nishume, J. D. Engel, and M. Yamamoto. 1994. Developmental stage- and spermatogenic cycle-specific expression of transcription factor GATA-1 in mouse Sertoli cells. Development 120:1759-1766[Abstract]. |
| 52. |
Yuan, H.,
N. Corbi,
C. Basilico, and L. Dailey.
1995.
Developmental-specific activity of the FGF-4 enhancer requires the synergistic action of Sox2 and Oct-3.
Genes Dev.
9:2635-2645 |
Molecular and Cellular Biology, November 1998, p. 6653-6665, Vol. 18, No. 11
0270-7306/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.
This article has been cited by other articles:
-
Bradford, S. T., Wilhelm, D., Bandiera, R., Vidal, V., Schedl, A., Koopman, P.
(2009). A cell-autonomous role for WT1 in regulating Sry in vivo. Hum Mol Genet
18: 3429-3438
[Abstract] [Full Text] -
Quek, S. I., Chan, W. K.
(2009). Transcriptional activation of zebrafish cyp11a1 promoter is dependent on the nuclear receptor Ff1b. J Mol Endocrinol
43: 121-130
[Abstract] [Full Text] -
Dupasquier, S., Abdel-Samad, R., Glazer, R. I., Bastide, P., Jay, P., Joubert, D., Cavailles, V., Blache, P., Quittau-Prevostel, C.
(2009). A new mechanism of SOX9 action to regulate PKC{alpha} expression in the intestine epithelium. J. Cell Sci.
122: 2191-2196
[Abstract] [Full Text] -
Bradford, S. T., Hiramatsu, R., Maddugoda, M. P., Bernard, P., Chaboissier, M.-C., Sinclair, A., Schedl, A., Harley, V., Kanai, Y., Koopman, P., Wilhelm, D.
(2009). The Cerebellin 4 Precursor Gene Is a Direct Target of SRY and SOX9 in Mice. Biol. Reprod.
80: 1178-1188
[Abstract] [Full Text] -
Beverdam, A., Svingen, T., Bagheri-Fam, S., Bernard, P., McClive, P., Robson, M., Khojasteh, M. B., Salehi, M., Sinclair, A. H, Harley, V. R, Koopman, P.
(2009). Sox9-dependent expression of Gstm6 in Sertoli cells during testis development in mice. Reproduction
137: 481-486
[Abstract] [Full Text] -
Yang, W.-H., Heaton, J. H., Brevig, H., Mukherjee, S., Iniguez-Lluhi, J. A., Hammer, G. D.
(2009). SUMOylation Inhibits SF-1 Activity by Reducing CDK7-Mediated Serine 203 Phosphorylation. Mol. Cell. Biol.
29: 613-625
[Abstract] [Full Text] -
Chang, H., Gao, F., Guillou, F., Taketo, M. M., Huff, V., Behringer, R. R.
(2008). Wt1 negatively regulates {beta}-catenin signaling during testis development. Development
135: 1875-1885
[Abstract] [Full Text] -
Mouillet, J.-F., Yan, X., Ou, Q., Jin, L., Muglia, L. J., Crawford, P. A., Sadovsky, Y.
(2008). DEAD-Box Protein-103 (DP103, Ddx20) Is Essential for Early Embryonic Development and Modulates Ovarian Morphology and Function. Endocrinology
149: 2168-2175
[Abstract] [Full Text] -
Wang, H., Leav, I., Ibaragi, S., Wegner, M., Hu, G.-f., Lu, M. L., Balk, S. P., Yuan, X.
(2008). SOX9 Is Expressed in Human Fetal Prostate Epithelium and Enhances Prostate Cancer Invasion. Cancer Res.
68: 1625-1630
[Abstract] [Full Text] -
Shoemaker, C. M., Queen, J., Crews, D.
(2007). Response of Candidate Sex-Determining Genes to Changes in Temperature Reveals Their Involvement in the Molecular Network Underlying Temperature-Dependent Sex Determination. Mol. Endocrinol.
21: 2750-2763
[Abstract] [Full Text] -
Clement, T. M, Anway, M. D, Uzumcu, M., Skinner, M. K
(2007). Regulation of the gonadal transcriptome during sex determination and testis morphogenesis: comparative candidate genes. Reproduction
134: 455-472
[Abstract] [Full Text] -
Bastide, P., Darido, C., Pannequin, J., Kist, R., Robine, S., Marty-Double, C., Bibeau, F., Scherer, G., Joubert, D., Hollande, F., Blache, P., Jay, P.
(2007). Sox9 regulates cell proliferation and is required for Paneth cell differentiation in the intestinal epithelium. JCB
178: 635-648
[Abstract] [Full Text] -
Park, S. Y., Tong, M., Jameson, J. L.
(2007). Distinct Roles for Steroidogenic factor 1 and Desert hedgehog Pathways in Fetal and Adult Leydig Cell Development. Endocrinology
148: 3704-3710
[Abstract] [Full Text] -
Guimont, P., Grondin, F., Dubois, C. M.
(2007). Sox9-dependent transcriptional regulation of the proprotein convertase furin. Am. J. Physiol. Cell Physiol.
293: C172-C183
[Abstract] [Full Text] -
Sumi, E., Iehara, N., Akiyama, H., Matsubara, T., Mima, A., Kanamori, H., Fukatsu, A., Salant, D. J., Kita, T., Arai, H., Doi, T.
(2007). SRY-Related HMG Box 9 Regulates the Expression of Col4a2 through Transactivating Its Enhancer Element in Mesangial Cells. Am. J. Pathol.
170: 1854-1864
[Abstract] [Full Text] -
Wilhelm, D., Hiramatsu, R., Mizusaki, H., Widjaja, L., Combes, A. N., Kanai, Y., Koopman, P.
(2007). SOX9 Regulates Prostaglandin D Synthase Gene Transcription in Vivo to Ensure Testis Development. J. Biol. Chem.
282: 10553-10560
[Abstract] [Full Text] -
Wang, H., McKnight, N. C., Zhang, T., Lu, M. L., Balk, S. P., Yuan, X.
(2007). SOX9 Is Expressed in Normal Prostate Basal Cells and Regulates Androgen Receptor Expression in Prostate Cancer Cells. Cancer Res.
67: 528-536
[Abstract] [Full Text] -
Wilhelm, D., Palmer, S., Koopman, P.
(2007). Sex Determination and Gonadal Development in Mammals. Physiol. Rev.
87: 1-28
[Abstract] [Full Text] -
Benoit, G., Cooney, A., Giguere, V., Ingraham, H., Lazar, M., Muscat, G., Perlmann, T., Renaud, J.-P., Schwabe, J., Sladek, F., Tsai, M.-J., Laudet, V.
(2006). International Union of Pharmacology. LXVI. Orphan Nuclear Receptors. Pharmacol. Rev.
58: 798-836
[Abstract] [Full Text] -
Sugawara, T., Sakuragi, N., Minakami, H.
(2006). CREM confers cAMP responsiveness in human steroidogenic acute regulatory protein expression in NCI-H295R cells rather than SF-1/Ad4BP.. J Endocrinol
191: 327-337
[Abstract] [Full Text] -
Gao, F., Maiti, S., Alam, N., Zhang, Z., Deng, J. M., Behringer, R. R., Lecureuil, C., Guillou, F., Huff, V.
(2006). The Wilms tumor gene, Wt1, is required for Sox9 expression and maintenance of tubular architecture in the developing testis. Proc. Natl. Acad. Sci. USA
103: 11987-11992
[Abstract] [Full Text] -
Johnsen, I. K, Slawik, M., Shapiro, I., Hartmann, M. F, Wudy, S. A, Looyenga, B. D, Hammer, G. D, Reincke, M., Beuschlein, F.
(2006). Gonadectomy in mice of the inbred strain CE/J induces proliferation of sub-capsular adrenal cells expressing gonadal marker genes.. J Endocrinol
190: 47-57
[Abstract] [Full Text] -
Wissmuller, S., Kosian, T., Wolf, M., Finzsch, M., Wegner, M.
(2006). The high-mobility-group domain of Sox proteins interacts with DNA-binding domains of many transcription factors.. Nucleic Acids Res
34: 1735-1744
[Abstract] [Full Text] -
Barrionuevo, F., Bagheri-Fam, S., Klattig, J., Kist, R., Taketo, M. M., Englert, C., Scherer, G.
(2006). Homozygous Inactivation of Sox9 Causes Complete XY Sex Reversal in Mice. Biol. Reprod.
74: 195-201
[Abstract] [Full Text] -
Chen, W.-Y., Juan, L.-J., Chung, B.-c.
(2005). SF-1 (Nuclear Receptor 5A1) Activity Is Activated by Cyclic AMP via p300-Mediated Recruitment to Active Foci, Acetylation, and Increased DNA Binding. Mol. Cell. Biol.
25: 10442-10453
[Abstract] [Full Text] -
Baron, D., Houlgatte, R., Fostier, A., Guiguen, Y.
(2005). Large-Scale Temporal Gene Expression Profiling During Gonadal Differentiation and Early Gametogenesis in Rainbow Trout. Biol. Reprod.
73: 959-966
[Abstract] [Full Text] -
Yao, H. H-C, Capel, B.
(2005). Temperature, Genes, and Sex: a Comparative View of Sex Determination in Trachemys scripta and Mus musculus. J Biochem
138: 5-12
[Abstract] [Full Text] -
Guigon, C. J., Coudouel, N., Mazaud-Guittot, S., Forest, M. G., Magre, S.
(2005). Follicular Cells Acquire Sertoli Cell Characteristics after Oocyte Loss. Endocrinology
146: 2992-3004
[Abstract] [Full Text] -
Park, S. Y., Meeks, J. J., Raverot, G., Pfaff, L. E., Weiss, J., Hammer, G. D., Jameson, J. L.
(2005). Nuclear receptors Sf1 and Dax1 function cooperatively to mediate somatic cell differentiation during testis development. Development
132: 2415-2423
[Abstract] [Full Text] -
Jay, P., Berta, P., Blache, P.
(2005). Expression of the Carcinoembryonic Antigen Gene Is Inhibited by SOX9 in Human Colon Carcinoma Cells. Cancer Res.
65: 2193-2198
[Abstract] [Full Text] -
Park, S. Y., Jameson, J. L.
(2005). Minireview: Transcriptional Regulation of Gonadal Development and Differentiation. Endocrinology
146: 1035-1042
[Abstract] [Full Text] -
Shirai, T., Miyagi, S., Horiuchi, D., Okuda-Katayanagi, T., Nishimoto, M., Muramatsu, M., Sakamoto, Y., Nagata, M., Hagiwara, K., Okuda, A.
(2005). Identification of an Enhancer That Controls Up-regulation of Fibronectin during Differentiation of Embryonic Stem Cells into Extraembryonic Endoderm. J. Biol. Chem.
280: 7244-7252
[Abstract] [Full Text] -
Wilson, M. J., Jeyasuria, P., Parker, K. L., Koopman, P.
(2005). The Transcription Factors Steroidogenic Factor-1 and SOX9 Regulate Expression of Vanin-1 during Mouse Testis Development. J. Biol. Chem.
280: 5917-5923
[Abstract] [Full Text] -
Komatsu, T., Mizusaki, H., Mukai, T., Ogawa, H., Baba, D., Shirakawa, M., Hatakeyama, S., Nakayama, K. I., Yamamoto, H., Kikuchi, A., Morohashi, K.-i.
(2004). Small Ubiquitin-Like Modifier 1 (SUMO-1) Modification of the Synergy Control Motif of Ad4 Binding Protein/Steroidogenic Factor 1 (Ad4BP/SF-1) Regulates Synergistic Transcription between Ad4BP/SF-1 and Sox9. Mol. Endocrinol.
18: 2451-2462
[Abstract] [Full Text] -
Britt, K. L., Stanton, P. G., Misso, M., Simpson, E. R., Findlay, J. K.
(2004). The Effects of Estrogen on the Expression of Genes Underlying the Differentiation of Somatic Cells in the Murine Gonad. Endocrinology
145: 3950-3960
[Abstract] [Full Text] -
Moniot, B., Biau, S., Faure, S., Nielsen, C. M., Berta, P., Roberts, D. J., de Santa Barbara, P.
(2004). SOX9 specifies the pyloric sphincter epithelium through mesenchymal-epithelial signals. Development
131: 3795-3804
[Abstract] [Full Text] -
Blache, P., van de Wetering, M., Duluc, I., Domon, C., Berta, P., Freund, J.-N., Clevers, H., Jay, P.
(2004). SOX9 is an intestine crypt transcription factor, is regulated by the Wnt pathway, and represses the CDX2 and MUC2 genes. JCB
166: 37-47
[Abstract] [Full Text] -
Hirvonen-Santti, S. J., Sriraman, V., Anttonen, M., Savolainen, S., Palvimo, J. J., Heikinheimo, M., Richards, J. S., Janne, O. A.
(2004). Small Nuclear RING Finger Protein Expression during Gonad Development: Regulation by Gonadotropins and Estrogen in the Postnatal Ovary. Endocrinology
145: 2433-2444
[Abstract] [Full Text] -
Chaboissier, M.-C., Kobayashi, A., Vidal, V. I. P., Lutzkendorf, S., van de Kant, H. J. G., Wegner, M., de Rooij, D. G., Behringer, R. R., Schedl, A.
(2004). Functional analysis of Sox8 and Sox9 during sex determination in the mouse. Development
131: 1891-1901
[Abstract] [Full Text] -
MacLaughlin, D. T., Donahoe, P. K.
(2004). Sex Determination and Differentiation. NEJM
350: 367-378
[Full Text] -
LaVoie, H. A.
(2003). The Role of GATA in Mammalian Reproduction. Exp. Biol. Med.
228: 1282-1290
[Abstract] [Full Text] -
Cheung, M., Briscoe, J.
(2003). Neural crest development is regulated by the transcription factor Sox9. Development
130: 5681-5693
[Abstract] [Full Text] -
Hossain, A., Saunders, G. F.
(2003). Role of Wilms Tumor 1 (WT1) in the Transcriptional Regulation of the Mullerian-Inhibiting Substance Promoter. Biol. Reprod.
69: 1808-1814
[Abstract] [Full Text] -
Tremblay, J. J., Viger, R. S.
(2003). A Mutated Form of Steroidogenic Factor 1 (SF-1 G35E) That Causes Sex Reversal in Humans Fails to Synergize with Transcription Factor GATA-4. J. Biol. Chem.
278: 42637-42642
[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] -
Remenyi, A., Lins, K., Nissen, L. J., Reinbold, R., Scholer, H. R., Wilmanns, M.
(2003). Crystal structure of a POU/HMG/DNA ternary complex suggests differential assembly of Oct4 and Sox2 on two enhancers. Genes Dev.
17: 2048-2059
[Abstract] [Full Text] -
Harley, V. R., Clarkson, M. J., Argentaro, A.
(2003). The Molecular Action and Regulation of the Testis-Determining Factors, SRY (Sex-Determining Region on the Y Chromosome) and SOX9 [SRY-Related High-Mobility Group (HMG) Box 9]. Endocr. Rev.
24: 466-487
[Abstract] [Full Text] -
Schepers, G., Wilson, M., Wilhelm, D., Koopman, P.
(2003). SOX8 Is Expressed during Testis Differentiation in Mice and Synergizes with SF1 to Activate the Amh Promoter in Vitro. J. Biol. Chem.
278: 28101-28108
[Abstract] [Full Text] -
Bernard, P., Tang, P., Liu, S., Dewing, P., Harley, V. R., Vilain, E.
(2003). Dimerization of SOX9 is required for chondrogenesis, but not for sex determination. Hum Mol Genet
12: 1755-1765
[Abstract] [Full Text] -
Sock, E., Pagon, R. A., Keymolen, K., Lissens, W., Wegner, M., Scherer, G.
(2003). Loss of DNA-dependent dimerization of the transcription factor SOX9 as a cause for campomelic dysplasia. Hum Mol Genet
12: 1439-1447
[Abstract] [Full Text] -
Martinez, A., Val, P., Sahut-Barnola, I., Aigueperse, C., Veyssiere, G., Lefrancois-Martinez, A.-M.
(2003). Steroidogenic Factor-1 Controls the Aldose Reductase akr1b7 Gene Promoter in Transgenic Mice through an Atypical Binding Site. Endocrinology
144: 2111-2120
[Abstract] [Full Text] -
Lukas-Croisier, C., Lasala, C., Nicaud, J., Bedecarras, P., Kumar, T. R., Dutertre, M., Matzuk, M. M., Picard, J.-Y., Josso, N., Rey, R.
(2003). Follicle-Stimulating Hormone Increases Testicular Anti-Mullerian Hormone (AMH) Production through Sertoli Cell Proliferation and a Nonclassical Cyclic Adenosine 5'-Monophosphate-Mediated Activation of the AMH Gene. Mol. Endocrinol.
17: 550-561
[Abstract] [Full Text] -
Anttonen, M., Ketola, I., Parviainen, H., Pusa, A.-K., Heikinheimo, M.
(2003). FOG-2 and GATA-4 Are Coexpressed in the Mouse Ovary and Can Modulate Mullerian-Inhibiting Substance Expression. Biol. Reprod.
68: 1333-1340
[Abstract] [Full Text] -
Huang, W., Lu, N., Eberspaecher, H., de Crombrugghe, B.
(2002). A New Long Form of c-Maf Cooperates with Sox9 to Activate the Type II Collagen Gene. J. Biol. Chem.
277: 50668-50675
[Abstract] [Full Text] -
Gurates, B., Sebastian, S., Yang, S., Zhou, J., Tamura, M., Fang, Z., Suzuki, T., Sasano, H., Bulun, S. E.
(2002). WT1 and DAX-1 Inhibit Aromatase P450 Expression in Human Endometrial and Endometriotic Stromal Cells. J. Clin. Endocrinol. Metab.
87: 4369-4377
[Abstract] [Full Text] -
BRITT, K. L., KERR, J., O'DONNELL, L., JONES, M. E. E., DRUMMOND, A. E., DAVIS, S. R., SIMPSON, E. R., FINDLAY, J. K.
(2002). Estrogen regulates development of the somatic cell phenotype in the eutherian ovary. FASEB J.
16: 1389-1397
[Abstract] [Full Text] -
Gasca, S., Canizares, J., de Santa Barbara, P., Mejean, C., Poulat, F., Berta, P., Boizet-Bonhoure, B.
(2002). A nuclear export signal within the high mobility group domain regulates the nucleocytoplasmic translocation of SOX9 during sexual determination. Proc. Natl. Acad. Sci. USA
99: 11199-11204
[Abstract] [Full Text] -
Zhou, R., Bonneaud, N., Yuan, C.-X., de Santa Barbara, P., Boizet, B., Tibor, S., Scherer, G., Roeder, R. G., Poulat, F., Berta, P.
(2002). SOX9 interacts with a component of the human thyroid hormone receptor-associated protein complex. Nucleic Acids Res
30: 3245-3252
[Abstract] [Full Text] -
Wilhelm, D., Englert, C.
(2002). The Wilms tumor suppressor WT1 regulates early gonad development by activation of Sf1. Genes Dev.
16: 1839-1851
[Abstract] [Full Text] -
Hoyle, C., Narvaez, V., Alldus, G., Lovell-Badge, R., Swain, A.
(2002). Dax1 Expression Is Dependent on Steroidogenic Factor 1 in the Developing Gonad. Mol. Endocrinol.
16: 747-756
[Abstract] [Full Text] -
Shen, J. H.-C., Ingraham, H. A.
(2002). Regulation of the Orphan Nuclear Receptor Steroidogenic Factor 1 by Sox Proteins. Mol. Endocrinol.
16: 529-540
[Abstract] [Full Text] -
Ohe, K., Lalli, E., Sassone-Corsi, P.
(2002). A direct role of SRY and SOX proteins in pre-mRNA splicing. Proc. Natl. Acad. Sci. USA
10.1073/pnas.022645899v1
[Abstract] [Full Text] -
Adams, I. R., McLaren, A.
(2002). Sexually dimorphic development of mouse primordial germ cells: switching from oogenesis to spermatogenesis. Development
129: 1155-1164
[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] -
Ikeda, Y., Nagai, A., Ikeda, M.-a., Hayashi, S.
(2002). Increased Expression of Mullerian-Inhibiting Substance Correlates with Inhibition of Follicular Growth in the Developing Ovary of Rats Treated with E2 Benzoate. Endocrinology
143: 304-312
[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] -
Teixeira, J., Maheswaran, S., Donahoe, P. K.
(2001). Mullerian Inhibiting Substance: An Instructive Developmental Hormone with Diagnostic and Possible Therapeutic Applications. Endocr. Rev.
22: 657-674
[Abstract] [Full Text] -
Tremblay, J. J., Robert, N. M., Viger, R. S.
(2001). Modulation of Endogenous GATA-4 Activity Reveals Its Dual Contribution to Mullerian Inhibiting Substance Gene Transcription in Sertoli Cells. Mol. Endocrinol.
15: 1636-1650
[Abstract] [Full Text] -
MacLaughlin, D. T., Teixeira, J., Donahoe, P. K.
(2001). Perspective: Reproductive Tract Development--New Discoveries and Future Directions. Endocrinology
142: 2167-2172
[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] -
Tremblay, J. J., Viger, R. S.
(2001). GATA Factors Differentially Activate Multiple Gonadal Promoters through Conserved GATA Regulatory Elements. Endocrinology
142: 977-986
[Abstract] [Full Text] -
de Santa Barbara, P., Méjean, C., Moniot, B., Malclès, M.-H., Berta, P., Boizet-Bonhoure, B.
(2001). Steroidogenic Factor-1 Contributes to the Cyclic-Adenosine Monophosphate Down-Regulation of Human SRY Gene Expression. Biol. Reprod.
64: 775-783
[Abstract] [Full Text] -
Pincas, H., Amoyel, K., Counis, R., Laverrière, J.-N.
(2001). Proximal cis-Acting Elements, Including Steroidogenic Factor 1, Mediate the Efficiency of a Distal Enhancer in the Promoter of the Rat Gonadotropin-Releasing Hormone Receptor Gene. Mol. Endocrinol.
15: 319-337
[Abstract] [Full Text] -
Nef, S., Parada, L. F.
(2000). Hormones in male sexual development. Genes Dev.
14: 3075-3086
[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] -
Beranger, F., Mejean, C., Moniot, B., Berta, P., Vandromme, M.
(2000). Muscle Differentiation Is Antagonized by SOX15, a New Member of the SOX Protein Family. J. Biol. Chem.
275: 16103-16109
[Abstract] [Full Text] -
Frigeri, C., Tsao, J., Czerwinski, W., Schimmer, B. P.
(2000). Impaired Steroidogenic Factor 1 (NR5A1) Activity in Mutant Y1 Mouse Adrenocortical Tumor Cells. Mol. Endocrinol.
14: 535-544
[Abstract] [Full Text] -
Barnea, E., Bergman, Y.
(2000). Synergy of SF1 and RAR in Activation of Oct-3/4 Promoter. J. Biol. Chem.
275: 6608-6619
[Abstract] [Full Text] -
Watanabe, K., Clarke, T. R., Lane, A. H., Wang, X., Donahoe, P. K.
(2000). Endogenous expression of Mullerian inhibiting substance in early postnatal rat Sertoli cells requires multiple steroidogenic factor-1 and GATA-4-binding sites. Proc. Natl. Acad. Sci. USA
97: 1624-1629
[Abstract] [Full Text] -
Ito, M., Park, Y., Weck, J., Mayo, K. E., Jameson, J. L.
(2000). Synergistic Activation of the Inhibin {alpha}-Promoter by Steroidogenic Factor-1 and Cyclic Adenosine 3',5'-Monophosphate. Mol. Endocrinol.
14: 66-81
[Abstract] [Full Text] -
Teixeira, J., Kehas, D. J., Antun, R., Donahoe, P. K.
(1999). Transcriptional regulation of the rat Mullerian inhibiting substance type II receptor in rodent Leydig cells. Proc. Natl. Acad. Sci. USA
96: 13831-13838
[Abstract] [Full Text] -
Parma, P., Pailhoux, E., Cotinot, C.
(1999). Reverse Transcription-Polymerase Chain Reaction Analysis of Genes Involved in Gonadal Differentiation in Pigs. Biol. Reprod.
61: 741-748
[Abstract] [Full Text] -
Kawabe, K., Shikayama, T., Tsuboi, H., Oka, S., Oba, K., Yanase, T., Nawata, H., Morohashi, K.-i.
(1999). Dax-1 as One of the Target Genes of Ad4BP/SF-1. Mol. Endocrinol.
13: 1267-1284
[Abstract] [Full Text] -
Tremblay, J. J., Viger, R. S.
(1999). Transcription Factor GATA-4 Enhances Mullerian Inhibiting Substance Gene Transcription through a Direct Interaction with the Nuclear Receptor SF-1. Mol. Endocrinol.
13: 1388-1401
[Abstract] [Full Text] -
Swain, A., Lovell-Badge, R.
(1999). Mammalian sex determination: a molecular drama. Genes Dev.
13: 755-767
[Full Text] -
Ito, M., Achermann, J. C., Jameson, J. L.
(2000). A Naturally Occurring Steroidogenic Factor-1 Mutation Exhibits Differential Binding and Activation of Target Genes. J. Biol. Chem.
275: 31708-31714
[Abstract] [Full Text] -
Hossain, A., Saunders, G. F.
(2001). The Human Sex-determining Gene SRY Is a Direct Target of WT1. J. Biol. Chem.
276: 16817-16823
[Abstract] [Full Text] -
Preiss, S., Argentaro, A., Clayton, A., John, A., Jans, D. A., Ogata, T., Nagai, T., Barroso, I., Schafer, A. J., Harley, V. R.
(2001). Compound Effects of Point Mutations Causing Campomelic Dysplasia/Autosomal Sex Reversal upon SOX9 Structure, Nuclear Transport, DNA Binding, and Transcriptional Activation. J. Biol. Chem.
276: 27864-27872
[Abstract] [Full Text] -
Panda, D. K., Miao, D., Lefebvre, V., Hendy, G. N., Goltzman, D.
(2001). The Transcription Factor SOX9 Regulates Cell Cycle and Differentiation Genes in Chondrocytic CFK2 Cells. J. Biol. Chem.
276: 41229-41236
[Abstract] [Full Text] -
Ohe, K., Lalli, E., Sassone-Corsi, P.
(2002). A direct role of SRY and SOX proteins in pre-mRNA splicing. Proc. Natl. Acad. Sci. USA
99: 1146-1151
[Abstract] [Full Text]
| |||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Copyright © 2009 by the American Society for Microbiology. For an alternate route to Journals.ASM.org, visit: http://intl-journals.asm.org | More Info»