| HOME | HELP | FEEDBACK | SUBSCRIPTIONS | ARCHIVE | SEARCH | TABLE OF CONTENTS |
Unité de Recherche en Ontogénie et Reproduction Centre Hospitalier Universitaire de Québec Pavillon Centre Hospitalier de l’Université Laval Ste-Foy, Québec, Canada G1V 4G2
 |
ABSTRACT |
|---|
 TOP ABSTRACT INTRODUCTIONRESULTSDISCUSSIONMATERIALS AND METHODSREFERENCES |
|---|
|
INTRODUCTION |
|---|
|
TOPABSTRACTINTRODUCTIONRESULTSDISCUSSIONMATERIALS AND METHODSREFERENCES |
|---|
In addition to their critical roles in gonadal development, the SF-1, WT-1, SOX9, and SRY transcription factors also play equally important roles in the control of sex-specific gene expression. The best studied male-specific gene lying downstream of these factors in the male sex differentiation pathway is Müllerian inhibiting substance (MIS), which encodes a hormone produced by Sertoli cells of the fetal testis. MIS regulates male sexual differentiation by triggering the regression of the Müllerian ducts, the anlagen of the female reproductive tract, in genotypic XY males. MIS gene expression is sexually dimorphic. Sertoli cells begin to express MIS on embryonic day 12.5 (E12.5) in the mouse; expression is maintained throughout fetal development and then declines markedly after birth (12, 13). In contrast, MIS is apparently not expressed in the fetal and early postnatal ovary but can be detected in granulosa cells of the adult ovary (12, 13).
The identification of transacting factors involved in the regulation of the MIS gene has progressed rapidly in the past few years. The first gene shown to be crucial for MIS expression is the orphan nuclear receptor SF-1 (1, 14). Indeed, targeted disruption of the Ftz-F1 gene, which encodes SF-1, prevents Müllerian duct regression (1) and, accordingly, in recent transgenic studies Giuili et al. (14) have demonstrated that an intact SF-1-binding site is required for the sex-specific expression of MIS. Since SF-1 is expressed in many tissues where MIS is not, there has been an active search for potential mechanisms that could restrict MIS expression to the gonads. An attractive possibility, which has been frequently observed in other systems, is that gonad-specific MIS expression is the result of a combinatorial interaction between SF-1 and other transcriptional regulators. Indeed, the ability of SF-1 to activate MIS transcription has recently been shown to be enhanced through direct protein interactions with WT-1 and Sox9 (15, 16). In part, these SF-1/WT-1 and SF-1/Sox9 interactions have been helpful in our understanding of the cell-specific expression of the MIS gene. We have recently reported, however, the presence of GATA-4, a member of the GATA family of transcriptional regulators, in the developing gonads that could also play an integral role in the cell-specific and high level of MIS expression in the gonads (17). In the mouse, GATA-4 protein is abundant in the somatic cell population of the indifferent gonad just before the MIS gene is first turned on (17). Like MIS, GATA-4 expression later becomes restricted to the Sertoli cell lineage of the fetal testis and granulosa cells of the adult ovary (17). Moreover, the proximal MIS promoter contains a functional, species-conserved GATA element (17). Although other GATA factors have been shown to be expressed in the gonads, they are expressed after the MIS gene is markedly down-regulated (18).
The GATA factors, which bind the WGATAR consensus in the 5'-regulatory region of target genes, are presently an intensely studied group of transcriptional regulators due to their established roles in cell differentiation, organ development, and cell-specific gene expression in many systems (19, 20, 21, 22, 23, 24). The GATA family comprises six members (GATA-1 to -6) that can be divided into two subgroups: the hematopoietic (GATA-1 to -3) and the cardiac (GATA-4 to -6) groups. Although GATA factors have similar DNA-binding properties, they exhibit distinct spatial and developmental expression patterns and play essential, nonredundant functions (19, 20, 21, 22, 23, 24). Mounting evidence in the literature suggests that the functional specificity of the different GATA factors appears to be achieved, in part, via direct protein-protein interactions with other cell-restricted factors. Indeed, the zinc finger cofactor FOG interacts with GATA-1 to synergistically activate hematopoietic-specific gene expression (25). GATA-1, -2, and -3 have also been shown to interact with several other factors, including RBTN2, NF-E2, EKLF, TAL1, and Sp1, to regulate the activity of erythroid- and lymphoid-specific promoters and enhancers (26, 27, 28, 29, 30). Similarly, GATA-4 has recently been shown to directly interact and transcriptionally cooperate with the cardiac homeodomain protein Nkx2–5 to synergistically activate the atrial natriuretic peptide (ANF) promoter in the heart (31, 32).
The MIS promoter is a downstream target for GATA-4 and SF-1 in Sertoli cells (12, 14, 17). We used this promoter to examine whether both factors functionally cooperate to regulate gene expression in the gonads. We show here that coexpression of GATA-4 and SF-1 markedly enhances the activity of the MIS promoter and a panel of other SF-1-dependent targets. Moreover, we provide evidence that this synergy is the result of a novel interaction, both in vitro and in vivo, between SF-1 and the zinc finger region of GATA-4. Interestingly, synergy between GATA-4 and SF-1 did not absolutely require GATA binding to DNA. Thus, the present data not only provide new insights into the cascade of factors that control the sex-specific expression of the MIS gene but also reveals a potentially new mechanism for modulating SF-1 activity in tissues where both GATA factors and SF-1 are coexpressed.
|
RESULTS |
|---|
|
TOPABSTRACTINTRODUCTIONRESULTSDISCUSSIONMATERIALS AND METHODSREFERENCES |
|---|
, alignment of the proximal MIS promoter
from different species reveals the presence of three highly conserved
regions: a SRY-like element recently reported to bind the SRY-related
protein Sox9 (15), a binding site for the orphan nuclear receptor SF-1
(12), and a consensus GATA motif that we have recently shown to be
recognized by GATA-4 in Sertoli cells (17). Interestingly, the GATA and
SF-1 binding sites in the MIS promoter are 10 bp apart. This close
proximity raises the intriguing possibility that GATA-4 might represent
a novel cofactor for SF-1 in the sex-specific regulation of
MIS gene expression in the gonads. Although GATA-4 and SF-1
have been shown to regulate MIS promoter activity on their own (12, 14, 17), transcriptional synergism between the two factors has not been
reported in the gonads or other tissues where SF-1 and GATA factors are
coexpressed.
|
and Ref. 12). In contrast,
GATA-4 significantly activates the MIS promoter (Fig. 2A).
Surprisingly, the presence of both GATA-4 and SF-1 resulted in a
synergistic activation of the -180 bp MIS promoter (Fig. 2A, left panel). Synergy was also observed between GATA-4 and a
shorter SF-1 receptor (SF-1
LBD), which was deleted of its
ligand-binding domain (LBD), suggesting that synergistic activation of
the MIS promoter by GATA-4 and SF-1 does not require the LBD of SF-1
(Fig. 2A, middle panel). As control, no synergy was observed
on the minimal (-65 bp) MIS promoter, which does not contain GATA and
SF-1 binding sites (Fig. 2A, right panel).
|
, B and C). The first
consisted of three copies of an oligonucleotide containing the MIS SF-1
and GATA sites in their normal context fused to the unresponsive
minimal MIS promoter [(SF-1:GATA)3, Fig. 2B]. As observed
for the -180 bp MIS promoter, SF-1 remained a weak activator of this
reporter despite the presence of three SF-1 binding sites. Several
members of the GATA factor family were then tested for synergy with
SF-1 (Fig. 2B). Surprisingly, all GATA factors strongly synergized with
SF-1, suggesting that the mechanism underlying GATA/SF-1 synergy is
likely mediated via a domain common to all GATA factors. Indeed, we
have identified this domain to be the zinc finger region (see Fig. 7, discussed below). As shown in Fig. 2C, synergy between GATA-4 and SF-1
was not restricted to the GATA/SF-1 elements of the MIS promoter. The
-142 bp LHß promoter, which contains both SF-1 (33) and putative
GATA-binding sites, was also synergistically activated by GATA-4 and
SF-1 (Fig. 2C, left panel). Similarly, a synthetic reporter
containing two GATA elements and three copies of the SF-1-binding site
from the LHß promoter upstream of the minimal POMC promoter exhibited
strong synergism in the presence of both factors (Fig. 2C, middle
panel). As observed for the minimal MIS promoter, GATA-4 and SF-1
failed to activate the minimal POMC promoter, which lacks GATA- and
SF-1-binding sites (Fig. 2C, right panel). Thus, the synergy
between GATA-4 and SF-1 appears to be independent of promoter
context.
|
. The DNA-binding activity of
the GATA proteins was first assessed by gel shift assay. With the
exception of the two deletion mutants (internal and
N3, Fig. 3B, lanes 9 and 10) that remove the second zinc
finger, which has been shown to be essential for DNA binding (34, 35),
all GATA-4 proteins were expressed in the nucleus at similar levels, as
evidenced by their binding to the MIS GATA element (Fig. 3B). The
transcriptional properties of the GATA-4 deletion mutants were
subsequently tested in cotransfection experiments using a reporter
containing two highly responsive GATA motifs upstream of the minimal
MIS promoter. As shown in Fig. 3C, deletion of the entire N-terminal
domain up to the second zinc finger (N2) led to a 2-fold
increase in transcriptional activity when compared with the
wild-type GATA-4 protein. Further N-terminal deletion that
removed the second zinc finger (N3), or an internal
deletion that removed the second zinc finger but kept the first zinc
finger and nuclear localization signal intact
(internal), completely abolished transcriptional
activity. A recent study has reported that the second zinc finger of
GATA-4, in addition to the nuclear localization signal, is also
involved in the nuclear localization of the protein since its deletion
results in diffuse protein expression both in the nucleus and
cytoplasm (34). However, even when internal and
N3 were used at higher DNA concentrations to ensure
sufficient nuclear targeting of the proteins, these mutants still
failed to bind DNA and activate a GATA-dependent reporter (Fig. 3, B
and C). Interestingly, deletion of the C-terminal domain (from aa 333
to 440, C1) led to an important decrease in
transcriptional activity, suggesting that a potent activation domain
(AD) is present within this region of the GATA-4 protein. The remaining
activity (
25% of wild-type) represents another AD that has
previously been identified in the N-terminal region of GATA proteins
(34). Consistent with these observations, deletion of both the N- and
C-terminal domains (N1C1 and
N2C1) completely abrogated GATA
responsiveness (Fig. 3C). Taken together, these results indicate that
the GATA-4 protein contains two ADs that flank the DNA-binding domain,
the most potent of which is located in the C-terminal region.
|
, two GATA-4 domains proved to be
crucial for synergy with SF-1: the C-terminal AD and the second zinc
finger. The presence of both domains was required for synergy, since
deletion of either domain (as reflected in the C1 and
N3 mutants), abolished the GATA-4-dependent enhancement
of SF-1 activity (Fig. 4). Consistent with the requirement of these two
domains, deletion of the entire N-terminal domain and the first zinc
finger (N2) did not abrogate synergy with SF-1.
|
). Synergy between
GATA-4 and SF-1 was still observed on a smaller -107 bp MIS promoter
fragment which, like the -180 bp construct, retains both the GATA- and
SF-1-binding sites (Fig. 5A). A -83 bp MIS promoter fragment, which
conserves the GATA but removes the SF-1-binding site, was activated
solely by GATA-4 (Fig. 5B). No synergy was observed when both factors
were used in combination, indicating that a GATA motif by itself is not
sufficient for transcriptional synergism with SF-1. However,
coexpression of SF-1 with GATA-4 substantially repressed the
GATA-4-dependent activation of this promoter construct (Fig. 5B). This
repression likely represents squelching due to an interaction between
GATA-4 and SF-1, as previously reported for other transcription factors
(36, 37, 38, 39). Similar results were obtained using other GATA-dependent
promoters that lack SF-1 sites: the -114 bp cardiac B-type natriuretic
peptide (BNP) promoter, which contains GATA- but no SF-1-binding sites
(Fig. 5F), or two synthetic reporters containing two GATA motifs
upstream of either the minimal MIS (Fig. 5D) or POMC (Fig. 5E)
promoters. In contrast to the need for an SF-1 site, mutation of the
GATA motif (GATA 
GGTA) in the MIS promoter, which was previously
shown to abolish DNA binding and activation by GATA-4 (17), did not
prevent synergy between GATA-4 and SF-1 (Fig. 5C). The fact that SF-1
can synergize with GATA-4 without the latter binding to DNA suggests
that GATA-4/SF-1 synergism might be the result of a direct
protein-protein interaction between the two factors.
|
, in vitro translated
35S-labeled GATA-4 was specifically retained by an
immobilized maltose-binding protein (MBP)-SF-1 fusion protein (lane 2
in left panel). No labeled protein was retained, however,
when a MBP-LacZ
fusion was used as control (lane 3 in left
panel of Fig. 6A). This indicated that the GATA-4/SF-1 interaction
was not mediated by the MBP moiety of the MBP-SF-1 fusion protein.
Moreover, the direct interaction between GATA-4 and SF-1 was specific
since no interaction was observed when an unrelated
35S-labeled luciferase protein was used in the assay (Fig. 6A, right panel).
|
). In these experiments, three different
SF-1-dependent reporters were used: three copies of the SF-1 binding
site from the LHß promoter fused to the minimal POMC promoter
(left panel), two copies of the MIS SF-1 site upstream of
the minimal MIS promoter (middle panel), and five copies of
the SF-1 binding site from the steroid 21-hydroxylase promoter fused to
the minimal PRL promoter (right panel). As expected, SF-1,
but not GATA-4, activated the three reporters. However, when GATA-4 and
SF-1 were used in combination, GATA-4 significantly enhanced the
SF-1-dependent activation of all three constructs. Since these
SF-1-dependent reporters do not contain GATA motifs to permit GATA
binding, the synergy observed when both GATA-4 and SF-1 were present
indicates an in vivo interaction between the two
factors.
SF-1 Interacts with Either Zinc Finger of GATA-4
The domain(s) of the GATA-4 protein involved in the direct
interaction with SF-1 was initially mapped using in vitro
pull-down assays (Fig. 7A). N-terminal
deletions right up to the second zinc finger (N1,
N2) did not affect the ability of the mutant GATA-4
protein to interact with SF-1. Similarly, a deletion removing the
entire C-terminal domain (C1) did not prevent an
interaction with SF-1. Deletions that removed both zinc fingers of
GATA-4 (N3 or C2), however, abolished the
interaction with SF-1. Taken together, these data narrowed the
interaction domain to the zinc finger region of GATA-4, a region we
previously defined to be required for transcriptional synergy with SF-1
(Fig. 4). Indeed, both zinc fingers of GATA-4 were found to interact
with SF-1. The second zinc finger (N2C1),
however, appeared to bind more strongly to SF-1 than the first zinc
finger (internal).
The mapping of the GATA-4 interaction domain was confirmed in
vivo by cotransfection experiments using the SF-1-dependent
reporters previously described. As shown in Fig. 7B, we found that when
either the first (aa 201 to 266) or second (aa 242 to 301) zinc finger
region of GATA-4 was fused to the C-terminal AD required for synergy
(N2 and internal), a significant
enhancement of SF-1-dependent activity was observed. This enhancement
was not observed with the C-terminal domain alone (N3),
even when used at higher doses of DNA to ensure nuclear localization.
Thus, both zinc fingers of the GATA-4 protein can interact in
vivo with the nuclear receptor SF-1.
|
DISCUSSION |
|---|
|
TOPABSTRACTINTRODUCTIONRESULTSDISCUSSIONMATERIALS AND METHODSREFERENCES |
|---|
GATA-4/SF-1 Synergism Is Mediated via the Zinc Finger Region of
GATA-4
As depicted in Fig. 8A, the GATA-4
domain required for the physical interaction with SF-1 was mapped to
the DNA-binding region. A more detailed analysis of the GATA-4
interaction domain revealed that both GATA-4 zinc fingers interacted
with SF-1, resulting in transcriptional synergism in the presence of
the C-terminal AD of GATA-4 (Fig. 7). Since the DNA-binding domain is
highly conserved among the GATA factors, it was not surprising to
observe synergy (Fig. 2B) and physical interaction (data not shown)
between SF-1 and different GATA family members. Our data are consistent
with previous reports that have also identified the GATA zinc finger
region as a crucial domain involved in protein-protein interactions
with other transcription factors. For example, GATA-1, -2, and -3 have
all been shown to interact with the Krüppel family factors, Sp1
and EKLF, through their respective zinc finger regions (26). Similarly,
self-association of GATA-1 occurs through its zinc finger domains (42).
The C-terminal zinc finger of GATA-4 has also been recently shown to
interact with the cardiac homeoprotein Nkx2–5 (31, 32, 43), whereas
the interaction between GATA-1 and its cofactor FOG is mediated
exclusively through the N-terminal zinc finger of GATA-1 (25). Thus,
the zinc finger region of GATA proteins appears to be crucial for
protein-protein interaction, as illustrated in the present work
involving GATA-4 and SF-1.
|
We initially thought that synergy between GATA-4 and SF-1 would be
specific to the MIS promoter, given that the SF-1- and GATA-binding
sites are in close proximity to each other (Fig. 1). Remarkably, we
also observed strong synergistic activation by GATA-4 and SF-1 on a
variety of other natural and synthetic SF-1-dependent promoters such as
the LHß promoter (Fig. 2). The LHß promoter is a well characterized
downstream target for SF-1 in pituitary gonadotropes (57, 58).
Interestingly, the proximal LHß promoter contains an SF-1 binding
site (33) and a potential GATA motif. Moreover, GATA factors, including
GATA-4, are coexpressed with SF-1 in pituitary gonadotropes (59),
suggesting that transcriptional cooperation between GATA and SF-1 may
also be implicated in pituitary-specific gene expression. The fact that
synergy with SF-1 was also observed with other GATA family members
(Fig. 2B) and the fact that GATA/SF-1 synergy did not absolutely
require GATA binding to DNA (Figs. 5C, 6B, and 7B) are further
indications that transcriptional cooperation between GATA factors and
SF-1 may have broader implications. Thus, the functional interaction
between GATA-4 and SF-1, described here, appears to represent a more
generalized mechanism for the regulation of SF-1 target genes in
tissues where both SF-1 and GATA factors are coexpressed.
A Role for GATA-4 in Male Sex Differentiation
One of the critical steps in the establishment of the proper male
phenotype is the regression of the Müllerian ducts, which is
mediated through the action of MIS. At present, four factors have been
proposed to be involved in the sex-specific regulation of the
MIS gene: SF-1, WT-1, SOX9, and GATA-4. As shown in Fig. 8B, the onset of MIS gene expression occurs around E12.5 in the
mouse, shortly after testis differentiation. Based on solid evidence
recently reported in the literature (14, 16), MIS gene
expression in vivo appears to absolutely require the
presence of both SF-1 and WT-1. It is interesting to note, however,
that these two factors appear in the developing gonadal primordium of
the mouse (E9.5–10.5) somewhat before the MIS gene is
actually first turned on (E12.5) (13, 60, 61, 62). This observation
invariably suggests that another factor is likely missing at this time.
A recent report has proposed Sox9 to be this factor since it was shown
to interact with SF-1 (15). Several other lines of evidence, however,
appear to contradict this possibility. First, Sox9
expression (15), like SF-1 and WT-1, considerably
precedes that of MIS (13). Second, in granulosa cells of the
adult ovary, MIS is expressed in the absence of Sox9 (9, 13). Lastly, in the chick, MIS was shown to be expressed
before Sox9 (63). Taken together, these observations support
the existence of another factor that participates in the cell-specific
expression of the MIS gene. Our previous findings on the
sexually dimorphic expression pattern of the GATA-4 transcription
factor in the developing gonads, combined with the present functional
interaction data with SF-1 on the MIS promoter, support the notion
that GATA-4 is involved in MIS expression in
vivo. Indeed, abundant GATA-4 expression begins in the bipotential
gonad of the mouse on E11.5 (17), just before the detection of MIS
transcripts (13). Moreover, GATA-4, like MIS, is
expressed in granulosa cells of the adult mouse ovary (13, 17). Thus,
as presented in Fig. 8C, these data are consistent with a model in
which GATA-4 is an integral component of the combinatorial code of
factors required for MIS gene expression and, consequently,
male sex differentiation.
|
MATERIALS AND METHODS |
|---|
|
TOPABSTRACTINTRODUCTIONRESULTSDISCUSSIONMATERIALS AND METHODSREFERENCES |
|---|
GGTA) was obtained using the pALTER
site-directed mutagenesis system (Promega Corp., Madison,
WI); the mutation was confirmed by DNA sequencing. The -107 bp MIS
promoter was amplified by PCR using the -180 bp construct as template
and then cloned in the BamHI/HindIII site of the
luciferase expression vector pXP1, as previously described for the
other MIS promoter constructs (17). Since the pXP1 vector normally
contains two GATA motifs upstream of its multiple cloning site, all
pXP1 luciferase reporter constructs were modified to delete these
highly responsive GATA sites. This was achieved by replacing a
600-bp NdeI-BamHI fragment with a similar
fragment (minus the GATA sites) that was generated by PCR (forward
primer: 5'-TTCACACCGCATATGGTGCACT-3'; reverse primer:
5'-ACGGATCCAAGCTTACATTGATGAGTTTGGACAAAC-3') on
the promoterless pXP1 vector. Thus, the (GATA)2-MIS
luciferase reporter consists of the minimal (-65 bp) MIS promoter in
the unmodified pXP1 vector. The (SF-1:GATA)3-MIS luciferase
reporter was obtained by cloning three copies of a double-stranded MIS
SF-1:GATA element (sense oligonucleotide:
5'-GATCCAGGCACTGTCCCCCAAGGTCACCTTTGGTGTTGATAGGGGCGA-3'; antisense
oligonucleotide:
5'-GATCTCGCCCCTATCAACACCAAAGGTCACCTTGGGGGACAGTGCCTG-3') upstream of the
minimal MIS promoter in the modified pXP1 vector. Similarly, the
(SF-1)2-MIS luciferase reporter was obtained by cloning two
copies of the MIS SF-1 element (sense oligonucleotide:
5'-GATCCCCCAAGGTCACCTTTA-3'; antisense oligonucleotide:
5'-GATCTAAAGGTGACCTTGGGGG-3') in front of the minimal MIS promoter in
the modified pXP1 vector. The minimal POMC, (SF-1)3-POMC,
(GATA)2-POMC, (GATA)2/(SF-1)3-POMC,
and -142 bp LHß luciferase reporters were kind gifts of Jacques
Drouin (33, 64). In the POMC constructs, SF-1 refers to the SF-1
element found in the bovine LHß promoter (33). Again, the GATA motifs
in the POMC constructs given above come from an unmodified pXP1
luciferase reporter. GATA expression vectors, the -114 bp BNP
reporter, and certain GATA-4 deletion mutants (N1,
N2, C1, C2,
N1C1, N2C1) were
kindly provided by Mona Nemer. The remaining GATA-4 deletion mutants
(N3 and internal) were generated by PCR
on the wild-type GATA-4 cDNA and then cloned into the
XbaI/BamHI site of the pCG expression vector (31, 65). The forward primer for the N3 construct was
5'-CTTCTAGAGGGGTTCCCAGGCCTCTTGCA-3, and the reverse primer was
5'-CAGGATCCAAGTCCGAGCAGGAATTT-3'. The internal construct
was obtained by initially cloning the first zinc finger of GATA-4,
obtained by PCR, into pCG (forward primer:
5'-CTTCTAGACAACCCAATCTCGATATG-3'; reverse primer:
5'-ATGGATCCTTAGCTAGCCAGCCGGCGCTGAGGCTTGATGAGGGGC-3').
The latter was digested with NheI/BamHI (the
NheI site being provided by the reverse primer given above),
and the XbaI-BamHI C-terminal PCR fragment used
to produce N3 was cloned in frame to yield
internal. The SF-1 expression plasmid and
(SF-1)5-PRL reporter were generously provided by Keith
Parker. The (SF-1)5-PRL construct consists five copies of
the 21-hydroxylase SF-1 element cloned upstream of the minimal PRL
promoter (66).
Cell Culture and Transfections
African green monkey kidney CV-1 and murine L cells were grown
in DMEM supplemented with 10% newborn calf serum. Transfections were
done in 12-well plates using the calcium phosphate precipitation method
(67). CV-1 cells were plated at a density of 60,000 cells per well
24 h before transfection. Cell media were changed 12–16 h later,
and cells were harvested the next morning. Cells were lysed by adding
100 µl of lysis buffer (100 mM Tris-HCl, pH 7.9, 0.5%
Igepal (Sigma Chemical Co., St. Louis, MO), and 5
mM dithiothreitol) directly to the wells. Luciferase
activity (60 µl aliquot of lysate) was then assayed using an EG&G
Berthold (Bad Wildbad, Germany) LB 9507 luminometer. In synergy
experiments, optimal doses of SF-1 (10 ng/well) and GATA-4 (100
ng/well) expression vector were used. It is important to note that the
level of activation obtained when both GATA-4 and SF-1 were present
(synergy) could never be achieved by simply increasing the amount of
GATA-4 or SF-1 when used alone (independent activation). In fact, the
optimal dose of GATA-4 for maximum activation was 100 ng/well, which is
the same dose used in the synergy experiments. Thus, the synergy
observed when both factors are present indicates a true synergy. In all
experiments, the total amount of DNA was kept constant at 6 µg per
well using Sp64 (Promega Corp., Madison, WI) as carrier
DNA; several DNA preparations of the plasmids were used to ensure
reproducibility of the results. Data reported represent the average of
3 to 10 experiments, each done in duplicate.
Production of MBP Fusion Proteins
A recombinant MBP-SF-1 fusion protein was obtained by cloning
the entire coding region of murine SF-1 in frame with MBP using the
commercially available pMAL-c fusion protein vector (New England Biolabs, Inc., Mississauga, Ontario, Canada). The MBP-LacZ
fusion protein was provided by the wild-type pMAL-c vector. The two
fusion proteins constructs (MBP-SF-1 and MBP-LacZ) were introduced
into the Escherichia coli strain BL21, and fusion proteins
were produced by inducing the respective bacterial cultures with
isopropylthiogalactoside. Bacterial cultures were lysed by sonication,
and the fusion proteins were purified using an amylose resin (New England Biolabs, Inc.) as outlined by the manufacturer.
Protein-Protein Binding Assays
Protein-protein binding studies were done using
35S-labeled in vitro translated GATA-4 proteins
(wild-type and deletion mutants) and the purified MBP-SF-1 and
MBP-LacZ fusion proteins coupled to amylose resin. The
35S-labeled GATA-4 and luciferase proteins were obtained
using the TNT system from Promega Corp.; the amino acid
positions of the different GATA-4 proteins used are given in Fig. 3A.
Protein-protein interaction assays were done using 1 µg of MBP-fusion
protein and 10 µl of in vitro translated
35S-labeled protein essentially as described by Durocher
et al. (31). Bound complexes were separated by SDS-PAGE, and
retained proteins were revealed by autoradiography.
DNA-Binding Assays
Recombinant GATA proteins (wild-type and deletion mutants) were
obtained by transfecting L cells (which are devoid of GATA activity)
with the different GATA-4 expression plasmids described above. Nuclear
extracts were prepared 48 h after transfection by the procedure
outlined by Schreiber et al. (68). DNA-binding assays were
performed using a 32P-labeled double-stranded
oligonucleotide corresponding to the conserved MIS promoter GATA
element at -75 bp. Binding reactions and electrophoresis conditions
were as previously described (17).
|
ACKNOWLEDGMENTS |
|---|
|
FOOTNOTES |
|---|
This work was supported by a grant from the Medical Research Council of Canada to R.S.V.
Received for publication March 23, 1999. Accepted for publication May 13, 1999.
|
REFERENCES |
|---|
|
TOPABSTRACTINTRODUCTIONRESULTSDISCUSSIONMATERIALS AND METHODSREFERENCES |
|---|
-subunit gene in the placenta and
pituitary gland. Mol Cell Biol 14:5592–5602This article has been cited by other articles:
 |
|
 |
|
  L. J. Martin, N. Boucher, B. El-Asmar, and J. J. Tremblay cAMP-Induced Expression of the Orphan Nuclear Receptor Nur77 in MA-10 Leydig Cells Involves a CaMKI Pathway J Androl, March 1, 2009; 30(2): 134 - 145. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 B. El-Asmar, X. C Giner, and J. J Tremblay Transcriptional cooperation between NF-{kappa}B p50 and CCAAT/enhancer binding protein {beta} regulates Nur77 transcription in Leydig cells J. Mol. Endocrinol., February 1, 2009; 42(2): 131 - 138. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 W.-H. Yang, J. H. Heaton, H. Brevig, S. Mukherjee, J. A. Iniguez-Lluhi, and G. D. Hammer SUMOylation Inhibits SF-1 Activity by Reducing CDK7-Mediated Serine 203 Phosphorylation Mol. Cell. Biol., February 1, 2009; 29(3): 613 - 625. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 L. J. Martin, N. Boucher, C. Brousseau, and J. J. Tremblay The Orphan Nuclear Receptor NUR77 Regulates Hormone-Induced StAR Transcription in Leydig Cells through Cooperation with Ca2+/Calmodulin-Dependent Protein Kinase I Mol. Endocrinol., September 1, 2008; 22(9): 2021 - 2037. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 E. Lague and J. J. Tremblay Antagonistic Effects of Testosterone and the Endocrine Disruptor Mono-(2-Ethylhexyl) Phthalate on INSL3 Transcription in Leydig Cells Endocrinology, September 1, 2008; 149(9): 4688 - 4694. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 S. Vuorenoja, A. Rivero-Muller, A. J Ziecik, I. Huhtaniemi, J. Toppari, and N. A Rahman Targeted therapy for adrenocortical tumors in transgenic mice through their LH receptor by Hecate-human chorionic gonadotropin {beta} conjugate Endocr. Relat. Cancer, June 1, 2008; 15(2): 635 - 648. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
J.-F. Mouillet, X. Yan, Q. Ou, L. Jin, L. J. Muglia, P. A. Crawford, and Y. Sadovsky DEAD-Box Protein-103 (DP103, Ddx20) Is Essential for Early Embryonic Development and Modulates Ovarian Morphology and Function Endocrinology, May 1, 2008; 149(5): 2168 - 2175. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
R. S. Viger, S. M. Guittot, M. Anttonen, D. B. Wilson, and M. Heikinheimo Role of the GATA Family of Transcription Factors in Endocrine Development, Function, and Disease Mol. Endocrinol., April 1, 2008; 22(4): 781 - 798. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
C. Stocco, J. Kwintkiewicz, and Z. Cai Identification of regulatory elements in the Cyp19 proximal promoter in rat luteal cells J. Mol. Endocrinol., October 1, 2007; 39(4): 211 - 221. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 J. D. Shadley, K. Divakaran, K. Munson, R. N. Hines, K. Douglas, and D. G. McCarver Identification and Functional Analysis of a Novel Human CYP2E1 Far Upstream Enhancer Mol. Pharmacol., June 1, 2007; 71(6): 1630 - 1639. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
J. Kwintkiewicz, Z. Cai, and C. Stocco Follicle-Stimulating Hormone-Induced Activation of Gata4 Contributes in the Up-Regulation of Cyp19 Expression in Rat Granulosa Cells Mol. Endocrinol., April 1, 2007; 21(4): 933 - 947. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
A. Matsushita, S. Sasaki, Y. Kashiwabara, K. Nagayama, K. Ohba, H. Iwaki, H. Misawa, K. Ishizuka, and H. Nakamura Essential Role of GATA2 in the Negative Regulation of Thyrotropin {beta} Gene by Thyroid Hormone and Its Receptors Mol. Endocrinol., April 1, 2007; 21(4): 865 - 884. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
E. B. Dammer, A. Leon, and M. B. Sewer Coregulator Exchange and Sphingosine-Sensitive Cooperativity of Steroidogenic Factor-1, General Control Nonderepressed 5, p54, and p160 Coactivators Regulate Cyclic Adenosine 3',5'-Monophosphate-Dependent Cytochrome P450c17 Transcription Rate Mol. Endocrinol., February 1, 2007; 21(2): 415 - 438. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 D. Wilhelm, S. Palmer, and P. Koopman Sex Determination and Gonadal Development in Mammals Physiol Rev, January 1, 2007; 87(1): 1 - 28. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 S. Mazaud Guittot, A. Tetu, E. Legault, N. Pilon, D. W. Silversides, and R. S. Viger The Proximal Gata4 Promoter Directs Reporter Gene Expression to Sertoli Cells During Mouse Gonadal Development Biol Reprod, January 1, 2007; 76(1): 85 - 95. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 G. Benoit, A. Cooney, V. Giguere, H. Ingraham, M. Lazar, G. Muscat, T. Perlmann, J.-P. Renaud, J. Schwabe, F. Sladek, et al. International Union of Pharmacology. LXVI. Orphan Nuclear Receptors Pharmacol. Rev., December 1, 2006; 58(4): 798 - 836. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
B. D. Looyenga and G. D. Hammer Origin and Identity of Adrenocortical Tumors in Inhibin Knockout Mice: Implications for Cellular Plasticity in the Adrenal Cortex Mol. Endocrinol., November 1, 2006; 20(11): 2848 - 2863. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 K.-H. Song, Y.-Y. Park, H. J. Kee, C. Y. Hong, Y.-S. Lee, S.-W. Ahn, H.-J. Kim, K. Lee, H. Kook, I.-K. Lee, et al. Orphan Nuclear Receptor Nur77 Induces Zinc Finger Protein GIOT-1 Gene Expression, and GIOT-1 Acts as a Novel Corepressor of Orphan Nuclear Receptor SF-1 via Recruitment of HDAC2 J. Biol. Chem., June 9, 2006; 281(23): 15605 - 15614. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
M Anttonen, H Parviainen, A Kyronlahti, M Bielinska, D B Wilson, O Ritvos, and M Heikinheimo GATA-4 is a granulosa cell factor employed in inhibin-{alpha} activation by the TGF-{beta} pathway. J. Mol. Endocrinol., June 1, 2006; 36(3): 557 - 568. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 M. Bielinska, S. Kiiveri, H. Parviainen, S. Mannisto, M. Heikinheimo, and D. B. Wilson Gonadectomy-induced Adrenocortical Neoplasia in the Domestic Ferret (Mustela putorius furo) and Laboratory Mouse. Vet. Pathol., February 1, 2006; 43(2): 97 - 117. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
N. M. Robert, L. J. Martin, and J. J. Tremblay The Orphan Nuclear Receptor NR4A1 Regulates Insulin-Like 3 Gene Transcription in Leydig Cells Biol Reprod, February 1, 2006; 74(2): 322 - 330. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 M. Anttonen, L. Unkila-Kallio, A. Leminen, R. Butzow, and M. Heikinheimo High GATA-4 Expression Associates with Aggressive Behavior, whereas Low Anti-Mullerian Hormone Expression Associates with Growth Potential of Ovarian Granulosa Cell Tumors J. Clin. Endocrinol. Metab., December 1, 2005; 90(12): 6529 - 6535. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
M. F. Bouchard, H. Taniguchi, and R. S. Viger Protein Kinase A-Dependent Synergism between GATA Factors and the Nuclear Receptor, Liver Receptor Homolog-1, Regulates Human Aromatase (CYP19) PII Promoter Activity in Breast Cancer Cells Endocrinology, November 1, 2005; 146(11): 4905 - 4916. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
L. J. Martin, H. Taniguchi, N. M. Robert, J. Simard, J. J. Tremblay, and R. S. Viger GATA Factors and the Nuclear Receptors, Steroidogenic Factor 1/Liver Receptor Homolog 1, Are Key Mutual Partners in the Regulation of the Human 3{beta}-Hydroxysteroid Dehydrogenase Type 2 Promoter Mol. Endocrinol., September 1, 2005; 19(9): 2358 - 2370. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
M. Bielinska, E. Genova, I. Boime, H. Parviainen, S. Kiiveri, J. Leppaluoto, N. Rahman, M. Heikinheimo, and D. B. Wilson Gonadotropin-Induced Adrenocortical Neoplasia in NU/J Nude Mice Endocrinology, September 1, 2005; 146(9): 3975 - 3984. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
N. Huang, A. Dardis, and W. L. Miller Regulation of Cytochrome b5 Gene Transcription by Sp3, GATA-6, and Steroidogenic Factor 1 in Human Adrenal NCI-H295A Cells Mol. Endocrinol., August 1, 2005; 19(8): 2020 - 2034. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 H. H-C Yao and B. Capel Temperature, Genes, and Sex: a Comparative View of Sex Determination in Trachemys scripta and Mus musculus J. Biochem., July 1, 2005; 138(1): 5 - 12. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
L. J. Martin and J. J. Tremblay The Human 3{beta}-Hydroxysteroid Dehydrogenase/{Delta}5-{Delta}4 Isomerase Type 2 Promoter Is a Novel Target for the Immediate Early Orphan Nuclear Receptor Nur77 in Steroidogenic Cells Endocrinology, February 1, 2005; 146(2): 861 - 869. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
K. J. Saner, T. Suzuki, H. Sasano, J. Pizzey, C. Ho, J. F. Strauss III, B. R. Carr, and W. E. Rainey Steroid Sulfotransferase 2A1 Gene Transcription Is Regulated by Steroidogenic Factor 1 and GATA-6 in the Human Adrenal Mol. Endocrinol., January 1, 2005; 19(1): 184 - 197. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 J. K. Divine, L. J. Staloch, H. Haveri, C. M. Jacobsen, D. B. Wilson, M. Heikinheimo, and T. C. Simon GATA-4, GATA-5, and GATA-6 activate the rat liver fatty acid binding protein gene in concert with HNF-1{alpha} Am J Physiol Gastrointest Liver Physiol, November 1, 2004; 287(5): G1086 - G1099. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
T. Komatsu, H. Mizusaki, T. Mukai, H. Ogawa, D. Baba, M. Shirakawa, S. Hatakeyama, K. I. Nakayama, H. Yamamoto, A. Kikuchi, et al. 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., October 1, 2004; 18(10): 2451 - 2462. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
J. Qin, D.-m. Gao, Q.-F. Jiang, Q. Zhou, Y.-Y. Kong, Y. Wang, and Y.-H. Xie Prospero-Related Homeobox (Prox1) Is a Corepressor of Human Liver Receptor Homolog-1 and Suppresses the Transcription of the Cholesterol 7-{alpha}-Hydroxylase Gene Mol. Endocrinol., October 1, 2004; 18(10): 2424 - 2439. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
R. S. Viger, H. Taniguchi, N. M. Robert, and J. J. Tremblay The 25th Volume: Role of the GATA Family of Transcription Factors in Andrology J Androl, July 1, 2004; 25(4): 441 - 452. [Full Text] [PDF]
|
|
|
|
|
|
C. E. Fluck and W. L. Miller GATA-4 and GATA-6 Modulate Tissue-Specific Transcription of the Human Gene for P450c17 by Direct Interaction with Sp1 Mol. Endocrinol., May 1, 2004; 18(5): 1144 - 1157. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
H. Hiroi, L. K. Christenson, L. Chang, M. D. Sammel, S. L. Berger, and J. F. Strauss III Temporal and Spatial Changes in Transcription Factor Binding and Histone Modifications at the Steroidogenic Acute Regulatory Protein (StAR) Locus Associated with StAR Transcription Mol. Endocrinol., April 1, 2004; 18(4): 791 - 806. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
N. Lei and L. L. Heckert Gata4 Regulates Testis Expression of Dmrt1 Mol. Cell. Biol., January 1, 2004; 24(1): 377 - 388. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
A. J. Pask, D. J. Whitworth, C.-A. Mao, K.-J. Wei, N. Sankovic, J. A. M. Graves, G. Shaw, M. B. Renfree, and R. R. Behringer Marsupial Anti-Mullerian Hormone Gene Structure, Regulatory Elements, and Expression Biol Reprod, January 1, 2004; 70(1): 160 - 167. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
A. Hossain and G. F. Saunders Role of Wilms Tumor 1 (WT1) in the Transcriptional Regulation of the Mullerian-Inhibiting Substance Promoter Biol Reprod, December 1, 2003; 69(6): 1808 - 1814. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
J. J. Tremblay and R. S. Viger 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., October 24, 2003; 278(43): 42637 - 42642. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
Y.-W. Liu, W. Gao, H.-L. Teh, J.-H. Tan, and W.-K. Chan Prox1 Is a Novel Coregulator of Ff1b and Is Involved in the Embryonic Development of the Zebra Fish Interrenal Primordium Mol. Cell. Biol., October 15, 2003; 23(20): 7243 - 7255. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
P. Jimenez, K. Saner, B. Mayhew, and W. E. Rainey GATA-6 Is Expressed in the Human Adrenal and Regulates Transcription of Genes Required for Adrenal Androgen Biosynthesis Endocrinology, October 1, 2003; 144(10): 4285 - 4288. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
C. Y. Hong, J. H. Park, K. H. Seo, J.-M. Kim, S. Y. Im, J. W. Lee, H.-S. Choi, and K. Lee 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., September 1, 2003; 23(17): 6000 - 6012. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
M. Bielinska, H. Parviainen, S. B. Porter-Tinge, S. Kiiveri, E. Genova, N. Rahman, I. T. Huhtaniemi, L. J. Muglia, M. Heikinheimo, and D. B. Wilson Mouse Strain Susceptibility to Gonadectomy-Induced Adrenocortical Tumor Formation Correlates with the Expression of GATA-4 and Luteinizing Hormone Receptor Endocrinology, September 1, 2003; 144(9): 4123 - 4133. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
S. B. R. Jacobs, D. Coss, S. M. McGillivray, and P. L. Mellon Nuclear Factor Y and Steroidogenic Factor 1 Physically and Functionally Interact to Contribute to Cell-Specific Expression of the Mouse Follicle-Stimulating Hormone-{beta} Gene Mol. Endocrinol., August 1, 2003; 17(8): 1470 - 1483. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
G. Schepers, M. Wilson, D. Wilhelm, and P. Koopman SOX8 Is Expressed during Testis Differentiation in Mice and Synergizes with SF1 to Activate the Amh Promoter in Vitro J. Biol. Chem., July 18, 2003; 278(30): 28101 - 28108. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
J. J. Tremblay and R. S. Viger Transcription Factor GATA-4 Is Activated by Phosphorylation of Serine 261 via the cAMP/Protein Kinase A Signaling Pathway in Gonadal Cells J. Biol. Chem., June 6, 2003; 278(24): 22128 - 22135. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
I. Ketola, J. Toppari, T. Vaskivuo, R. Herva, J. S. Tapanainen, and M. Heikinheimo Transcription Factor GATA-6, Cell Proliferation, Apoptosis, and Apoptosis-Related Proteins Bcl-2 and Bax in Human Fetal Testis J. Clin. Endocrinol. Metab., April 1, 2003; 88(4): 1858 - 1865. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
C. Lukas-Croisier, C. Lasala, J. Nicaud, P. Bedecarras, T. R. Kumar, M. Dutertre, M. M. Matzuk, J.-Y. Picard, N. Josso, and R. Rey 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., April 1, 2003; 17(4): 550 - 561. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
M. Anttonen, I. Ketola, H. Parviainen, A.-K. Pusa, and M. Heikinheimo FOG-2 and GATA-4 Are Coexpressed in the Mouse Ovary and Can Modulate Mullerian-Inhibiting Substance Expression Biol Reprod, April 1, 2003; 68(4): 1333 - 1340. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
C. Gillio-Meina, Y. Y. Hui, and H. A. LaVoie GATA-4 and GATA-6 Transcription Factors: Expression, Immunohistochemical Localization, and Possible Function in the Porcine Ovary Biol Reprod, February 1, 2003; 68(2): 412 - 422. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
T. Suzuki, M. Kasahara, H. Yoshioka, K.-i. Morohashi, and K. Umesono LXXLL-Related Motifs in Dax-1 Have Target Specificity for the Orphan Nuclear Receptors Ad4BP/SF-1 and LRH-1 Mol. Cell. Biol., January 1, 2003; 23(1): 238 - 249. [Abstract] [Full Text]
|
|
|
|
|
|
J. J. Tremblay, F. Hamel, and R. S. Viger Protein Kinase A-Dependent Cooperation between GATA and CCAAT/Enhancer-Binding Protein Transcription Factors Regulates Steroidogenic Acute Regulatory Protein Promoter Activity Endocrinology, October 1, 2002; 143(10): 3935 - 3945. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
N. M. Robert, J. J. Tremblay, and R. S. Viger Friend of GATA (FOG)-1 and FOG-2 Differentially Repress the GATA-Dependent Activity of Multiple Gonadal Promoters Endocrinology, October 1, 2002; 143(10): 3963 - 3973. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
J. M. Dufour, R. V. Rajotte, and G. S. Korbutt Development of an In Vivo Model to Study Testicular Morphogenesis J Androl, September 1, 2002; 23(5): 635 - 644. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
S. Kiiveri, J. Liu, M. Westerholm-Ormio, N. Narita, D. B. Wilson, R. Voutilainen, and M. Heikinheimo Differential Expression of GATA-4 and GATA-6 in Fetal and Adult Mouse and Human Adrenal Tissue Endocrinology, August 1, 2002; 143(8): 3136 - 3143. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
R. Kerkela, S. Pikkarainen, T. Majalahti-Palviainen, H. Tokola, and H. Ruskoaho Distinct Roles of Mitogen-activated Protein Kinase Pathways in GATA-4 Transcription Factor-mediated Regulation of B-type Natriuretic Peptide Gene J. Biol. Chem., April 12, 2002; 277(16): 13752 - 13760. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
J. H.-C. Shen and H. A. Ingraham Regulation of the Orphan Nuclear Receptor Steroidogenic Factor 1 by Sox Proteins Mol. Endocrinol., March 1, 2002; 16(3): 529 - 540. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
Z. Zhang, A. Z. Wu, Z.-M. Feng, D. Mruk, C. Y. Cheng, and C.-L. C. Chen Gonadotropins, via cAMP, Negatively Regulate GATA-1 Gene Expression in Testicular Cells Endocrinology, March 1, 2002; 143(3): 829 - 836. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 K. L. Parker, D. A. Rice, D. S. Lala, Y. Ikeda, X. Luo, M. Wong, M. Bakke, L. Zhao, C. Frigeri, N. A. Hanley, et al. Steroidogenic Factor 1: an Essential Mediator of Endocrine Development Recent Prog. Horm. Res., January 1, 2002; 57(1): 19 - 36. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
Y. Ikeda, A. Nagai, M.-a. Ikeda, and S. Hayashi Increased Expression of Mullerian-Inhibiting Substance Correlates with Inhibition of Follicular Growth in the Developing Ovary of Rats Treated with E2 Benzoate Endocrinology, January 1, 2002; 143(1): 304 - 312. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 J. Teixeira, S. Maheswaran, and P. K. Donahoe Mullerian Inhibiting Substance: An Instructive Developmental Hormone with Diagnostic and Possible Therapeutic Applications Endocr. Rev., October 1, 2001; 22(5): 657 - 674. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
J. J. Tremblay, N. M. Robert, and R. S. Viger Modulation of Endogenous GATA-4 Activity Reveals Its Dual Contribution to Mullerian Inhibiting Substance Gene Transcription in Sertoli Cells Mol. Endocrinol., September 1, 2001; 15(9): 1636 - 1650. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
L. L. Heckert Activation of the Rat Follicle-Stimulating Hormone Receptor Promoter by Steroidogenic Factor 1 Is Blocked by Protein Kinase A and Requires Upstream Stimulatory Factor Binding to a Proximal E Box Element Mol. Endocrinol., May 1, 2001; 15(5): 704 - 715. [Abstract] [Full Text]
|
|
|
|
|
|
J. J. Tremblay and R. S. Viger Nuclear Receptor Dax-1 Represses the Transcriptional Cooperation Between GATA-4 and SF-1 in Sertoli Cells Biol Reprod, April 1, 2001; 64(4): 1191 - 1199. [Abstract] [Full Text]
|
|
|
|
|
|
J. J. Tremblay and R. S. Viger GATA Factors Differentially Activate Multiple Gonadal Promoters through Conserved GATA Regulatory Elements Endocrinology, March 1, 2001; 142(3): 977 - 986. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
H. Pincas, K. Amoyel, R. Counis, and J.-N. Laverrière 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., February 1, 2001; 15(2): 319 - 337. [Abstract] [Full Text]
|
|
|
|
|
|
J. Levallet, P. Koskimies, N. Rahman, and I. Huhtaniemi The Promoter of Murine Follicle-Stimulating Hormone Receptor: Functional Characterization and Regulation by Transcription Factor Steroidogenic Factor 1 Mol. Endocrinol., January 1, 2001; 15(1): 80 - 92. [Abstract] [Full Text]
|
|
|
|
|
|
Z.-M. Feng, A. Z. Wu, Z. Zhang, and C.-L. C. Chen GATA-1 and GATA-4 Transactivate Inhibin/Activin {beta}-B-Subunit Gene Transcription in Testicular Cells Mol. Endocrinol., November 1, 2000; 14(11): 1820 - 1835. [Abstract] [Full Text]
|
|
|
|
|
|
M. P. E. Laitinen, M. Anttonen, I. Ketola, D. B. Wilson, O. Ritvos, R. Butzow, and M. Heikinheimo Transcription Factors GATA-4 and GATA-6 and a GATA Family Cofactor, FOG-2, Are Expressed in Human Ovary and Sex Cord-Derived Ovarian Tumors J. Clin. Endocrinol. Metab., September 1, 2000; 85(9): 3476 - 3483. [Abstract] [Full Text]
|
|
|
|
|
|
C. R. Wooton-Kee and B. J. Clark Steroidogenic Factor-1 Influences Protein-Deoxyribonucleic Acid Interactions within the Cyclic Adenosine 3',5'-Monophosphate-Responsive Regions of the Murine Steroidogenic Acute Regulatory Protein Gene Endocrinology, April 1, 2000; 141(4): 1345 - 1355. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 K. Watanabe, T. R. Clarke, A. H. Lane, X. Wang, and P. K. Donahoe Endogenous expression of Mullerian inhibiting substance in early postnatal rat Sertoli cells requires multiple steroidogenic factor-1 and GATA-4-binding sites PNAS, February 15, 2000; 97(4): 1624 - 1629. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
J.-F. Pare, S. Roy, L. Galarneau, and L. Belanger The Mouse Fetoprotein Transcription Factor (FTF) Gene Promoter Is Regulated by Three GATA Elements with Tandem E Box and Nkx Motifs, and FTF in Turn Activates the Hnf3beta , Hnf4alpha , and Hnf1alpha Gene Promoters J. Biol. Chem., April 13, 2001; 276(16): 13136 - 13144. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
J. D. Molkentin The Zinc Finger-containing Transcription Factors GATA-4, -5, and -6. UBIQUITOUSLY EXPRESSED REGULATORS OF TISSUE-SPECIFIC GENE EXPRESSION J. Biol. Chem., December 8, 2000; 275(50): 38949 - 38952. [Full Text] [PDF]
|
|
| |||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
| HOME | HELP | FEEDBACK | SUBSCRIPTIONS | ARCHIVE | SEARCH | TABLE OF CONTENTS |
| Endocrinology | Endocrine Reviews | J. Clin. End. & Metab. |
| Molecular Endocrinology | Recent Prog. Horm. Res. | All Endocrine Journals |
).