| HOME | HELP | FEEDBACK | SUBSCRIPTIONS | ARCHIVE | SEARCH | TABLE OF CONTENTS |
Department of Obstetrics and Gynecology (J.A.P., G.G., Y.S.) and Pathology (P.A.C.), Washington University School of Medicine, St. Louis, Missouri 63110
 |
ABSTRACT |
|---|
 TOP ABSTRACT INTRODUCTIONRESULTSDISCUSSIONMATERIALS AND METHODSREFERENCES |
|---|
|
INTRODUCTION |
|---|
|
TOPABSTRACTINTRODUCTIONRESULTSDISCUSSIONMATERIALS AND METHODSREFERENCES |
|---|
Developmentally, SF-1 is expressed as early as E9 in the mouse urogenital ridge, before its morphological differentiation into adrenal and gonadal tissues (24). In the brain, SF-1 expression is detected in hypophyseal precursors, as well as in prosencephalic regions that give rise to the hypothalamus (24). To study the developmental and differentiated functions of SF-1 in vivo, Luo et al (25) and our group (26) generated SF-1 -/- mice. Intriguingly, these mice lacked gonads, resulting in the persistence of Mullerian structures in genotypically male and female mice. They also lacked adrenal glands, which led to their early neonatal death. Furthermore, SF-1 -/- mice demonstrated a developmental defect in the ventromedial hypothalamus, an area implicated in control of sexual behavior (27, 28). Together, these results demonstrate that SF-1 is expressed in tissues that are essential for reproductive and endocrine homeostasis and that it is required for intact development of the adrenal glands and gonads.
Members of the steroid receptor superfamily of proteins, like other transcription factors, are composed of modular functional domains (6). These include a DNA-binding domain, which is flanked by amino and carboxy termini. A ligand-independent activation function (AF-1) is commonly located at the amino terminus. The carboxy terminus harbors a hinge region and a ligand-binding domain, which commonly includes a second, ligand-dependent activation function (AF-2) domain (1, 29, 30, 31, 32). Transcriptional activity by these domains is either constitutive or ligand-dependent and determines the function of the nuclear receptors in development and differentiation.
Transcriptional activation domains are presumed to mediate an interaction with either components of the basal transcription machinery (33, 34, 35) or with cellular proteins distinct from basal transcription factors, termed coactivators. The interaction with coactivators may be of paramount importance in regulating physiological functions (2, 3). Several of these interacting proteins have been cloned recently and found to interact with the conserved AF-2 domain at the carboxy terminus of ligand-activated receptors. Several examples includes receptor-interacting protein 140, a coactivator of estrogen receptor [ER (36)], transcriptional intermediary factor 1, a regulator of ER, progesterone receptor (PR), retinoid X receptor (RXR), retinoic acid receptor (RAR), and vitamin D receptor (37), and cAMP response element (CREB)-binding protein (CBP), a coregulator of RXR and thyroid hormone receptor (TR) (38, 39). An additional coactivator, termed steroid receptor coactivator (SRC)-1, functions as a coactivator for ER, PR, glucocorticoid receptor (GR), TR, and RXR, as well as for several nonsteroid receptor activators such as Sp1 (40).
While the role of SF-1 in development and differentiation has been demonstrated, the mechanism of transcriptional activation by SF-1 has not been previously analyzed. Despite its ability to be activated by certain oxysterols (41), SF-1 differs from classic ligand-dependent steroid receptors, which have known high-affinity ligands that are essential for AF-2-mediated activation (42, 43, 44). Furthermore, SF-1 does not harbor a transcriptionally active amino terminus and, unlike many receptors that bind their response element as homo- or heterodimers, SF-1 binds DNA half-site sequences as a monomer (45). To identify the activation domains of SF-1, we initially characterized its AF-2 activation domain. While required for full SF-1 activity, this domain is not sufficient. An additional domain, located carboxy-terminal to the DNA-binding domain of SF-1, is also needed for gene activation. Moreover, we demonstrate that SRC-1 potentiates SF-1 activity in mammalian cells, and this interaction also requires both activation domains of SF-1.
|
RESULTS |
|---|
|
TOPABSTRACTINTRODUCTIONRESULTSDISCUSSIONMATERIALS AND METHODSREFERENCES |
|---|
), previously
identified as a conserved activation domain in several steroid
receptors (42, 43, 44, 46, 47, 48). This motif is composed of a glutamate,
preceded by a variable amino acid, flanked by two pairs of hydrophobic
amino acids. To test whether or not this hexamer confers
transcriptional activity to SF-1, we initially deleted the entire
hexamer and compared the transcriptional activity of the truncated
protein (SF-1
AF-2) to the activity of wild type SF-1, using the S25
reporter gene. As shown in Fig. 1B, removal of the 11 carboxy-terminal
amino acids of SF-1 diminished its activation capacity by 75% in both
JEG3 and CV-1 cells. Interestingly, removal of the terminal 183
residues (SF-1279) did not cause further diminution of the
activation capacity of SF-1 (Fig. 1B), suggesting that the AF-2 hexamer
is the most significant activation domain in this region of SF-1.
|
). As shown in
Fig. 1B, we found that mutation of either pair of hydrophobic amino
acids (M1, M4) diminished the activation capacity of SF-1 to a level
similar to SF-1AF-2, suggesting that each pair of hydrophobic
residues is essential for AF-2 function, as demonstrated for other
steroid receptors (42, 44, 46). In contrast, mutation of the conserved
glutamate (M3) had a weaker effect on the activation function of SF-1.
Not surprisingly, mutation of the nonconserved isoleucine (M2) had no
effect on activation by SF-1. A mutation of the glutamine at position
458 to either glutamate or alanine (M5) had no effect on activation by
SF-1. Similar results were obtained using a human P450scc
reporter construct (data not shown). These results indicate that,
unlike other steroid receptors such as ER, where mutation of either
pair of hydrophobic amino acids entirely abolish the activation
capacity of the protein, mutation of AF-2 activation hexamer (AF-2-AH)
or even a complete deletion of the hexamer renders SF-1 partly active,
albeit at a low level (20–30% of wild type SF-1). This suggests the
presence of an additional activation domain in SF-1.
To confirm that mutations in AF-2-AH did not alter the ability of
SF-1 to bind its response element half-site, we tested the capacity of
wild type and mutant SF-1 constructs to bind a double-stranded
oligonucleotide probe that contains an SF-1 response element (45). As
shown in Fig. 1C, we observed a comparable level of DNA binding by wild
type and mutant receptors in an electromobility shift assay. Taken
together, these results indicate that the AF-2-AH is an important
activation domain of SF-1.
The AF-2-AH Is Required for SF-1-Dependent Induction of
Steroidogenic Phenotype
Because SF-1 is required for development and differentiation
of steroidogenic tissues, we determined whether or not AF-2-AH is
important for this action of SF-1. For this purpose we used embryonic
stem (ES) cells, which differentiate into steroid-producing cells when
expressing SF-1 via stable transfection (49). Upon selection of
SF-1-transfected ES cells, we found that they expressed cytochrome
P450scc, a key enzyme in steroidogenic pathways (Fig. 2A), and released progesterone into the
medium (Fig. 2B). Remarkably, neither of these phenotypic changes was
observed when the SF-1 M4 mutant of AF-2-AH (Fig. 1A) was introduced
(Fig. 2A-B). These data suggest that SF-1’s AF-2-AH is required for
its physiological function in induction of a steroidogenic
phenotype.
|
) and tested for its
transcriptional activation capacity in JEG3 cells using a reporter
(GKI) that contains GAL4- binding sites. As shown in Fig. 3B, the
activity of GAL4-SF-1428 was similar to that of GAL4 alone,
suggesting that the AF-2-AH containing SF-1428 was not
functional as an independent activation domain. We obtained a similar
result when longer fragments of SF-1 (residues 279–462 or 245–465)
were fused to GAL4. In contrast, we observed a strong activation of the
GAL4 reporter gene by GAL4-SF-1187 and
GAL4-SF-1119. As control, we identified a similar
activation by GAL4-ER282-595 (which contains the
carboxy-terminal region of ER) in the presence of E2. Thus,
in addition to AF-2-AH, there is a second critical component of SF-1
activation function, located between residues 187–245. Next, to
determine whether AF-2-AH is required for the activity of this domain,
we tested for the activation capacity of
GAL4-SF-1119AF-2, or GAL4-SF-1119mAF-2,
which contain a mutation in the terminal pair of hydrophobic residues
within AF-2-AH (M4, Fig. 1A). The activation of the GAL4 reporter by
either mutant was diminished by 70–80% (Fig. 3B). Taken together,
these results indicate that AF-2–AH is required for the activation
capacity of SF-1, but is not sufficient for this function, and requires
an additional activation domain, which is located further upstream in
the carboxy-terminal domain of SF-1 (between residues 187–245).
|
). This
increase was similar to that seen with a fusion of GAL4 and the
carboxy-terminal region (residues 282–595) of ER (not shown). This
result indicates that SRC-1 interacts with the carboxy-terminal,
autonomously active region of SF-1.
|
), along with the SRC-1-VP16 vector, and
found that the interaction between SRC-1 and SF-1 absolutely requires
an intact AF-2-AH (Fig. 4B). Similarly, to test whether the proximal
activation domain is required for the interaction between SF-1 and
SRC-1, we transfected CV-1 cells with SRC-1-VP16, along with a series
of SF-1 fragments, fused to GAL4. We observed that the interaction of
SF-1 with SRC-1 in the two-hybrid system was abrogated when residues
between 187–245 were deleted from the SF-1 construct (Fig. 4C).
Together, these results indicate that both AF-2-AH and the proximal
activation domain (between residues 187–245) are required for the
activation capacity of SF-1, as well as for interaction with the
coactivator SRC-1. The relative positions of these domains are
indicated in Fig. 4D.
To determine whether these activation domains of SF-1 are required for
potentiation by SRC-1, we transiently transfected CV-1 cells with SF-1,
in the presence or absence of human SRC-1. For a positive control we
used ER (and its cognate reporter, Vit2P36-LUC), as SRC-1
has been previously shown to synergize with ER in regulation of
estrogen-responsive reporter genes (50). As shown in Fig. 5A, the addition of SRC-1 potentiated the
transcriptional activity of holo-SF-1 on its reporter construct S25,
while SRC-1 alone had no effect on promoter activity. Therefore, SRC-1
is a coactivator for SF-1. Next, we determined whether AF-2-AH and the
proximal activation domain are both required for potentiation of SF-1
activity by SRC-1. Using GAL41-147 fusions of the SF-1
carboxy terminus we found that the same SF-1 domains required for
autonomous activation capacity, and for two-hybrid interaction with
SRC-1-VP16, were also required for potentiation by intact SRC-1 (Fig. 5B). Therefore, the transcriptional activity of SF-1 depends on both
the proximal activation domain and AF-2-AH. The interaction of these
domains with SRC-1 may provide the mechanism for regulation of SF-1
signaling by SRC-1 in vivo.
|
|
DISCUSSION |
|---|
|
TOPABSTRACTINTRODUCTIONRESULTSDISCUSSIONMATERIALS AND METHODSREFERENCES |
|---|
Although SF-1’s AF-2-AH is conserved among other steroid receptors
(42, 43, 44, 46, 47, 48), there are two features that distinguish the AF-2-AH
in SF-1: 1) Unlike the AF-2 of other ligand-dependent steroid receptors
(such as ER) where mutations of either pair of hydrophobic amino acids
within AF-2-AH entirely abolished the transcriptional activity of the
protein, mutations in SF-1’s AF-2-AH only diminished its activity to
25–30% of wild-type SF-1. 2) A replacement of the conserved glutamate
with glutamine completely disrupts the activity of the chick
T3R
(43), yet a similar replacement in SF-1 only
diminished its activity by 50%. Interestingly, a replacement of this
pivotal residue with alanine had no effect on the activation capacity
of ER (42). As shown for other steroid receptors, none of these
mutations altered the DNA- binding capacity of SF-1. These differences
between SF-1 and ligand-dependent steroid receptors may be related to
the ligand independence of SF-1. Alternatively, these differences may
specify the interaction of AF-2-AH with other domains within SF-1 or
with a distinct repertoire of coactivators.
While essential for transcriptional regulation by SF-1, the AF-2-AH of
SF-1 does not exhibit an independent activation domain when fused to a
GAL4 DNA-binding domain. In contrast, a region of 35 amino acids that
span the AF-2 hexamer of T3R exhibits a strong
transcriptional activity when fused to GAL4 (43). Our results establish
that the AF-2-AH of SF-1 is necessary, but insufficient, for
transcriptional activation, and domains located further upstream
(residues 187–245) are essential for the transcriptional activity of
SF-1. Several steroid receptors contain additional activation domains
amino-terminal to the ligand-binding domain. For example, two proximal
activation domains (
2, 3) were identified in TR, and
homologous sequences have been found in RAR and RXR (46). Similarly, a
proximal transcriptional activation function resides between residues
181–310 in the SF-1 homolog xFF1rA (51). These activation domains are
capable of independent transcriptional activation when fused to
GAL41-147. The proximal activation domain of SF-1 is also
capable of activating a reporter gene even in the absence of the
AF-2-AH, albeit at a markedly reduced potency. These results suggest
that AF-2-AH and the proximal activation domains synergize in their
activation function. This synergy may be achieved through
intramolecular interaction between complementary domains or may furnish
a platform for complex interaction with coactivators. Importantly, the
proximal activation domain of SF-1 (amino acids 187–245) does not
appear to share significant homology with any protein other than the
closest relatives of SF-1, such as LRH-1 (FTF, PHR) and xFF1rA.
It has been proposed that coactivators are needed for bridging between
the steroid receptor and components of the basal transcriptional
machinery (52). This has been shown for RAR, which requires a E1A-like
factor to interact with transcription factor IID (53). Similarly, the
SWI/SNF family of yeast proteins are components of the basal
transcription machinery that enhance transcription by the steroid
receptors (54). SRC-1 is a recently identified coactivator for several
steroid receptors that can associate with nuclear receptor-CBP
complexes (see below), but the exact mechanism by which it potentiates
transcriptional activation by steroid receptors is presently unknown.
SRC-1 interacts with PR and ER and enhances the transcriptional
activation of PR, ER, TR, GR, and RXR in the presence of their
respective agonists (40, 52). It forms a family of proteins with
ER-associated protein 160, which interacts with ER, RARß, and RXR
(55), as well as GR-interacting protein 1 (transcriptional intermediary
factor 2), a protein that interacts with GR, ER, AR, RAR, and RXR in a
ligand-dependent fashion in yeast and mammalian cells (56, 57). Unlike
the SRC-1 association with most of these receptors, the interaction
between SF-1 and SRC-1 may be ligand-independent, as seen with some
steroid receptor coactivators (58). Our results indicate that SRC-1
potentiates the activation function of SF-1 in mammalian cells. The
AF-2-AH of SF-1 is required for SF-1 interaction with SRC-1, as the
interaction is abolished in the presence of a mutated AF-2-AH. However,
AF-2-AH is not sufficient for this interaction and requires an
additional, amino-terminal region of SF-1, located between residues
187–245. These data suggest that SRC-1 requires the correct
conformation of the two activation domains to interact with SF-1,
allowing signaling to the basal machinery or to additional
coactivators. Interestingly, a GAL4-SF-1119 construct that
is devoid of functional AF-2-AH does not interact with SRC-1, yet it
retains some transcriptional activity (Fig. 3B). This suggests that
additional coactivators may interact with proximal regions of SF-1.
The two-hybrid data presented herein demonstrate a functional interaction between SF-1 and SRC-1. However, our data do not prove that the interaction of the two proteins is direct, and it is possible that a complex of coactivating proteins is required for the functional interaction between SF-1 and SRC-1 to occur. Indeed, the coactivator CBP/p300 has been shown to participate in nuclear receptor-SRC-1 complexes and, in this case, could bridge SF-1 to SRC-1 (39, 59, 60, 61). Nevertheless, two lines of evidence argue for the direct interaction between SF-1 and SRC-1 in our two-hybrid assays. First, the interactive fragment of SRC-1 we employed for our two-hybrid assays contains none of the CBP/p300 interactive region (60). Second, using residues 1–450 of CBP in an analogous two-hybrid assay, we found that SF-1 and CBP do not interact in CV-1 cells (data not shown). Thus, because CBP/p300 does not appear to directly interact with SF-1, endogenous CBP probably does not bridge SF-1 to SRC-1 in the CV-1 two-hybrid assay. While CBP and SF-1 do not directly interact in CV-1 cells, it is still possible that an SF-1-SRC-1 interaction permits a ternary complex among SF-1, SRC-1, and CBP in vivo.
That SRC-1 plays a role in SF-1-mediated differentiation or function of steroidogenic cells remains to be determined. This is plausible, as a naturally occurring mutation within the AF-2-AH of human TRß disrupts interaction with SRC-1, implying that SRC-1 transduces the biological activity of TR (62). The results presented here demonstrate a clear correlation between the activation function of SF-1 and its interaction with SRC-1, implying that SRC-1 may be a primary determinant of SF-1 activity.
|
MATERIALS AND METHODS |
|---|
|
TOPABSTRACTINTRODUCTIONRESULTSDISCUSSIONMATERIALS AND METHODSREFERENCES |
|---|
AF-2 by placing a stop codon flanked by a 3'-EcoRI
site downstream from residue 451, thereby terminating SF-1 immediately
upstream from the AF-2-AH. The amplified EcoRI fragment was
cloned into pBS-KS and pCMV-Neo as described above for full-length
SF-1. CMV-SF-1279 was generated by SalI digestion of
pBS-revSF-1 and subcloning the EcoRI fragment of the
truncated SF-1 cDNA back into pBS, then using the KpnI and
XbaI sites as detailed above to clone into pCMV-Neo.
For mutagenesis of the SF-1’s AF-2-AH we used an inverse PCR-based
site-directed mutagenesis, using pBS-SF-1 as a template. The forward
primer harbored the mutations depicted in Fig. 1A and abutted a reverse
primer that encoded a wild type sequence. Using T4 kinase we
phosphorylated the reverse primer, then used KlenTaq (63) in ten cycles
of amplification with the following parameters: 94 C for 0.5 min, 56 C
for 2 min, 72 C for 5 min. The product was treated with 200 µg/ml
proteinase K for 30 min at 56 C, then phenol-chloroform-extracted,
ethanol-precipitated, treated with Pfu polymerase (Stratagene, La
Jolla, CA) at 37 C for 30 min, then ligated overnight. To remove the
original template, the ligation mix was treated with DpnI
for 1 h and transformed into XL-1 blue bacteria. The authenticity
of the mutant constructs was confirmed using the dideoxynucleotide
sequencing method on an Applied Biosystems (Foster City, CA) model 373A
DNA sequencer. All SF-1 constructs used for transfection of ES cells
were cloned as an EcoRI fragment (from pBS) into the vector
pCAGGS, which uses the cytomegalovirus immediate early enhancer and the
chicken ß-actin promoter and first intron enhancer, as described
elsewhere (49).
We used PCR to generate fusion proteins between the DNA-binding domain of GAL4 (1–147) and carboxy-terminal fragments of SF-1. Utilizing a BamHI-linked forward primer, a T3 reverse primer, and pBS-revSF-1 as a template, we amplified the desired fragments of SF-1 (residues 428–462, 279–462, 245–462, 187–462, 119–462, and 119–452), digested with BamHI and HindIII, and cloned downstream from GAL4 in pM2 vector (64). The correct frame of each chimeric protein was verified by sequencing. GAL4-ER282-595 was cloned in a similar fashion, using pBS-ER (a gift from Stuart Adler, Washington University) as a template.
A human SRC-1 expression vector (pBK-SRC1) was kindly provided by M. J. Tsai and B. W. O’Malley (Baylor College of Medicine, Houston, TX). To generate the SRC-1-VP16 fusion we amplified the receptor-interacting region of SRC-1 [corresponding to amino-acids 857-1061 (see Refs. 40 and 57); amino acids 1243-carboxy-terminus (see 60 ], and cloned it in frame upstream of the activation domain of VP16 (amino acids 413–440).
The SF-1 reporter construct (S25) contains the PRL minimal promoter
downstream from two SF-1-binding elements (TCAAGGTCA) separated by five
nucleotides, upstream from the luciferase gene. The
hP450scc-luciferase construct, which includes 2327 bp of
the human cytochrome P450scc promoter (65), was kindly
provided by W. L. Miller (University of California, San
Francisco). The GAL4 reporter gene (GKI), which contains five GAL4-
binding sites upstream of an E1B-TATA box, linked to luciferase,
was kindly provided by P. Webb and P. J. Kushner
(University of California, San Francisco). The ER reporter
construct (Vit2P36-LUC) was previously described (66).
Cell Culture and Transfection
CV-1 cells are maintained in DMEM that contains 10% FBS and
antibiotics at 37 C and 10% CO2. JEG3 cells are maintained
in MEM that contains 10% FBS and antibiotics at 37 C and 5%
CO2. All tissue culture reagents were obtained from the
Tissue Culture Support Facility, Washington University School of
Medicine (St. Louis, MO). RW4 mouse strain 129/SvJ embryonic stem (ES)
cells are maintained on a feeder layer of murine embryonic fibroblasts,
as described elsewhere (49).
One day before transfection, CV-1 or JEG3 cells were plated in either six- well plates at a density of 125,000 cells per well, or in 12-well plates at a density of 60,000 cells per well. Four hours before transfection the standard growth medium was replaced by fresh DMEM that contained additives as above, and the cells were incubated in 10% CO2 at 37 C. Transfection was performed using the standard calcium-phosphate precipitation method previously described (67) in six- and 12-well plates using 2.5 and 0.6 µg total DNA, respectively, which included 0.1 µg CMV ß-gal plasmid (to normalize for cell viability and transfection efficiency). Luciferase assay was performed 40–48 h after transfection. Cells were lysed in a lysis buffer that contains 50 mM Tris-2-(4-morpholino)-ethane sulfonic acid (pH 7.8), 1 mM dithiothreitol, and 1% Triton X-100. Lysates were assayed for luciferase using a luminometer (Monolight 2010 Analytical Luminescence Laboratory, San Diego, CA), and for ß-GAL using a 96-well plate reader (anthos htIII, Anthos Labtec, Salzburg, Austria). All experiments were performed in duplicate and repeated at least three times. Results (mean ±SD), normalized to ß-GAL activity, were expressed as relative luciferase units. In experiments in which ER was used we used phenol-red free medium with serum that contains negligible levels of E2.
For stable transfection of ES cells, 5 x 106 cells
were electroporated with 25 µg plasmid DNA as described elsewhere
(49). Transfected cells were plated on 100-mm tissue culture dishes
coated with a confluent layer of murine embryonic fibroblasts, and 300
µg/ml G418 or 1 µg/ml puromycin were administered the following day
for 5 days. On the sixth day, colonies were picked and expanded. For
stimulation experiments, ES cells were plated at 5 x
104 cells per gelatinized well of 24-well plates in 0.5 ml
ES media. After 1 day, cells were given 5 µg/ml
20-hydroxycholesterol in the presence or absence of 1 mM
8-bromo-cAMP.
Electromobility Shift Assay
Wild type or mutant SF-1 cDNA was transcribed and translated
from a pBSKS expression vector, using a TnT reticulocyte lysate system
(Promega, Madison, WI). A double-stranded oligonucleotide (100 ng) that
contained the SF-1 response element [TCAAGGTCA in tandem (45)], was
end-labeled by 20 µCi [
-32P]ATP, using
polynucleotide kinase. For each binding reaction 4 µl of the
translation mixture were mixed with 1 ng labeled probe in a binding
buffer (68) and incubated for 30 min at 25 C. Each binding reaction was
loaded onto a 5% polyacrylamide gel, run in 0.5x TBE buffer at 150 V
for 3 h. The gel was then dried and exposed to a PhosphorImager
(Molecular Dynamics, Sunnyvale, CA) screen or film.
RNA Analysis and Progesterone Measurement
RNA was extracted by the guanidinium thiocyanate-acid phenol
method (69). Total RNA (15 µg) was electrophoresed on a denaturing
gel of 1% agarose and 1.5% formaldehyde and blotted onto Zeta-Probe
GT membranes (Bio-Rad, Hercules, CA). The blot was probed with a
P450scc probe (a PCR product that spans the first 300 bp of
the mouse coding sequence), which was labeled with
[-32P]-dCTP, as previously described (70). Membranes
were hybridized for 16 h at 45 C, then washed with 0.2x SSC
(1xSSC is 0.15 M NaCl, and 0.075 M sodium
citrate) and 0.1% SDS for 20 min at 68 C, and analyzed by
PhosphorImager.
For progesterone assay, media were harvested 24 h after the addition of hormones, and progesterone was determined by RIA using the Coat-a-Count system (Diagnostic Products Corporation, Los Angeles, CA). Inter- and intraassay coefficients of variation were 5.1% and 2.6%, respectively.
|
ACKNOWLEDGMENTS |
|---|
|
FOOTNOTES |
|---|
This work was supported, in part, by NIH Grant HD-34110 and the Berlex Scholar Award (both to Y.S.).
Received for publication May 7, 1997. Revision received July 7, 1997. Accepted for publication July 17, 1997.
|
REFERENCES |
|---|
|
TOPABSTRACTINTRODUCTIONRESULTSDISCUSSIONMATERIALS AND METHODSREFERENCES |
|---|
4 activation domain of the thyroid hormone receptor is
required for release of the putative corepressor(s) necessary for
transcriptional silencing. Mol Cell Biol 15:76–86
bacteriophage templates. Proc Natl
Acad Sci USA 91:2216–2220
This article has been cited by other articles:
 |
|
 |
|
  H. A. LaVoie and S. R. King Transcriptional Regulation of Steroidogenic Genes: STARD1, CYP11A1 and HSD3B Experimental Biology and Medicine, August 1, 2009; 234(8): 880 - 907. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 B. Xu, W.-H. Yang, I. Gerin, C.-D. Hu, G. D. Hammer, and R. J. Koenig Dax-1 and Steroid Receptor RNA Activator (SRA) Function as Transcriptional Coactivators for Steroidogenic Factor 1 in Steroidogenesis Mol. Cell. Biol., April 1, 2009; 29(7): 1719 - 1734. [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]
|
|
|
|
 |
|
 A. E. Lewis, M. Rusten, E. A. Hoivik, E. L. Vikse, M. L. Hansson, A. E. Wallberg, and M. Bakke Phosphorylation of Steroidogenic Factor 1 Is Mediated by Cyclin-Dependent Kinase 7 Mol. Endocrinol., January 1, 2008; 22(1): 91 - 104. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
M. Doghman, T. Karpova, G. A. Rodrigues, M. Arhatte, J. De Moura, L. R. Cavalli, V. Virolle, P. Barbry, G. P. Zambetti, B. C. Figueiredo, et al. Increased Steroidogenic Factor-1 Dosage Triggers Adrenocortical Cell Proliferation and Cancer Mol. Endocrinol., December 1, 2007; 21(12): 2968 - 2987. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 W. Zheng, J. Yang, Q. Jiang, Z. He, and L. M Halvorson Liver receptor homologue-1 regulates gonadotrope function J. Mol. Endocrinol., February 1, 2007; 38(2): 207 - 219. [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]
|
|
|
|
|
|
S. H. Mellon, S. R. Bair, C. Depoix, J.-L. Vigne, N. B. Hecht, and P. B. Brake Translin Coactivates Steroidogenic Factor-1-Stimulated Transcription Mol. Endocrinol., January 1, 2007; 21(1): 89 - 105. [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]
|
|
|
|
 |
|
 T. N. Parakh, J. A. Hernandez, J. C. Grammer, J. Weck, M. Hunzicker-Dunn, A. J. Zeleznik, and J. H. Nilson Follicle-stimulating hormone/cAMP regulation of aromatase gene expression requires beta-catenin PNAS, August 15, 2006; 103(33): 12435 - 12440. [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]
|
|
|
|
|
|
J. Weck and K. E. Mayo Switching of NR5A Proteins Associated with the Inhibin {alpha}-Subunit Gene Promoter after Activation of the Gene in Granulosa Cells Mol. Endocrinol., May 1, 2006; 20(5): 1090 - 1103. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 J. N. Winnay, J. Xu, B. W. O'Malley, and G. D. Hammer Steroid Receptor Coactivator-1-Deficient Mice Exhibit Altered Hypothalamic-Pituitary-Adrenal Axis Function Endocrinology, March 1, 2006; 147(3): 1322 - 1332. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
J. N. Winnay and G. D. Hammer Adrenocorticotropic Hormone-Mediated Signaling Cascades Coordinate a Cyclic Pattern of Steroidogenic Factor 1-Dependent Transcriptional Activation Mol. Endocrinol., January 1, 2006; 20(1): 147 - 166. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
W.-Y. Chen, L.-J. Juan, and B.-c. Chung 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., December 1, 2005; 25(23): 10442 - 10453. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 S. N Kelly, T J. McKenna, and L. S Young Coregulatory protein-orphan nuclear receptor interactions in the human adrenal cortex J. Endocrinol., July 1, 2005; 186(1): 33 - 42. [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]
|
|
|
|
|
|
P.-L. Xu, Y.-Q. Liu, S.-F. Shan, Y.-Y. Kong, Q. Zhou, M. Li, J.-P. Ding, Y.-H. Xie, and Y. Wang Molecular Mechanism for the Potentiation of the Transcriptional Activity of Human Liver Receptor Homolog 1 by Steroid Receptor Coactivator-1 Mol. Endocrinol., August 1, 2004; 18(8): 1887 - 1905. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 R. V. Correa, S. Domenice, N. C. Bingham, A. E. C. Billerbeck, W. E. Rainey, K. L. Parker, and B. B. Mendonca A Microdeletion in the Ligand Binding Domain of Human Steroidogenic Factor 1 Causes XY Sex Reversal without Adrenal Insufficiency J. Clin. Endocrinol. Metab., April 1, 2004; 89(4): 1767 - 1772. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
J.-F. Mouillet, C. Sonnenberg-Hirche, X. Yan, and Y. Sadovsky p300 Regulates the Synergy of Steroidogenic Factor-1 and Early Growth Response-1 in Activating Luteinizing Hormone-{beta} Subunit Gene J. Biol. Chem., February 27, 2004; 279(9): 7832 - 7839. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
B. Borud, G. Mellgren, J. Lund, and M. Bakke Cloning and Characterization of a Novel Zinc Finger Protein that Modulates the Transcriptional Activity of Nuclear Receptors Mol. Endocrinol., November 1, 2003; 17(11): 2303 - 2319. [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]
|
|
|
|
|
|
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]
|
|
|
|
|
|
B. M. Gummow, J. N. Winnay, and G. D. Hammer Convergence of Wnt Signaling and Steroidogenic Factor-1 (SF-1) on Transcription of the Rat Inhibin {alpha} Gene J. Biol. Chem., July 11, 2003; 278(29): 26572 - 26579. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
M. Maira, C. Martens, E. Batsche, Y. Gauthier, and J. Drouin Dimer-Specific Potentiation of NGFI-B (Nur77) Transcriptional Activity by the Protein Kinase A Pathway and AF-1-Dependent Coactivator Recruitment Mol. Cell. Biol., February 1, 2003; 23(3): 763 - 776. [Abstract] [Full Text]
|
|
|
|
|
|
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]
|
|
|
|
|
|
X. Yan, J.-F. Mouillet, Q. Ou, and Y. Sadovsky A Novel Domain within the DEAD-Box Protein DP103 Is Essential for Transcriptional Repression and Helicase Activity Mol. Cell. Biol., January 1, 2003; 23(1): 414 - 423. [Abstract] [Full Text]
|
|
|
|
|
|
M. Desclozeaux, I. N. Krylova, F. Horn, R. J. Fletterick, and H. A. Ingraham Phosphorylation and Intramolecular Stabilization of the Ligand Binding Domain in the Nuclear Receptor Steroidogenic Factor 1 Mol. Cell. Biol., October 15, 2002; 22(20): 7193 - 7203. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
F. Gizard, B. Lavallee, F. DeWitte, E. Teissier, B. Staels, and D. W. Hum The Transcriptional Regulating Protein of 132 kDa (TReP-132) Enhances P450scc Gene Transcription through Interaction with Steroidogenic Factor-1 in Human Adrenal Cells J. Biol. Chem., October 11, 2002; 277(42): 39144 - 39155. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
P. Melamed, M. Koh, P. Preklathan, L. Bei, and C. Hew Multiple Mechanisms for Pitx-1 Transactivation of a Luteinizing Hormone beta Subunit Gene J. Biol. Chem., July 12, 2002; 277(29): 26200 - 26207. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
B. Borud, T. Hoang, M. Bakke, A. L. Jacob, J. Lund, and G. Mellgren The Nuclear Receptor Coactivators p300/CBP/Cointegrator-Associated Protein (p/CIP) and Transcription Intermediary Factor 2 (TIF2) Differentially Regulate PKA-Stimulated Transcriptional Activity of Steroidogenic Factor 1 Mol. Endocrinol., April 1, 2002; 16(4): 757 - 773. [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]
|
|
|
|
|
|
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]
|
|
|
|
|
|
Q. Ou, J.-F. Mouillet, X. Yan, C. Dorn, P. A. Crawford, and Y. Sadovsky The DEAD Box Protein DP103 Is a Regulator of Steroidogenic Factor-1 Mol. Endocrinol., January 1, 2001; 15(1): 69 - 79. [Abstract] [Full Text]
|
|
|
|
 |
|
 L Zhao, M Bakke, Y Krimkevich, L. Cushman, A. Parlow, S. Camper, and K. Parker Steroidogenic factor 1 (SF1) is essential for pituitary gonadotrope function Development, January 1, 2001; 128(2): 147 - 154. [Abstract] [PDF]
|
|
|
|
|
|
D. Boerboom, N. Pilon, R. Behdjani, D. W. Silversides, and J. Sirois 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, December 1, 2000; 141(12): 4647 - 4656. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
U. B. Kaiser, L. M. Halvorson, and M. T. Chen Sp1, Steroidogenic Factor 1 (SF-1), and Early Growth Response Protein 1 (Egr-1) Binding Sites Form a Tripartite Gonadotropin-Releasing Hormone Response Element in the Rat Luteinizing Hormone-{beta} Gene Promoter: an Integral Role for SF-1 Mol. Endocrinol., August 1, 2000; 14(8): 1235 - 1245. [Abstract] [Full Text]
|
|
|
|
|
|
C. Frigeri, J. Tsao, W. Czerwinski, and B. P. Schimmer Impaired Steroidogenic Factor 1 (NR5A1) Activity in Mutant Y1 Mouse Adrenocortical Tumor Cells Mol. Endocrinol., April 1, 2000; 14(4): 535 - 544. [Abstract] [Full Text]
|
|
|
|
|
|
W. Xie, H. Hong, N. N. Yang, R. J. Lin, C. M. Simon, M. R. Stallcup, and R. M. Evans Constitutive Activation of Transcription and Binding of Coactivator by Estrogen-Related Receptors 1 and 2 Mol. Endocrinol., December 1, 1999; 13(12): 2151 - 2162. [Abstract] [Full Text]
|
|
|
|
|
|
L.-A. Li, E. F-L. Chiang, J.-C. Chen, N.-C. Hsu, Y.-J. Chen, and B.-c. Chung Function of Steroidogenic Factor 1 Domains in Nuclear Localization, Transactivation, and Interaction with Transcription Factor TFIIB and c-Jun Mol. Endocrinol., September 1, 1999; 13(9): 1588 - 1598. [Abstract] [Full Text]
|
|
|
|
|
|
H. Hong, L. Yang, and M. R. Stallcup Hormone-independent Transcriptional Activation and Coactivator Binding by Novel Orphan Nuclear Receptor ERR3 J. Biol. Chem., August 6, 1999; 274(32): 22618 - 22626. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
R. Ahlgren, G. Suske, M. R. Waterman, and J. Lund Role of Sp1 in cAMP-dependent Transcriptional Regulation of the Bovine CYP11A Gene J. Biol. Chem., July 2, 1999; 274(27): 19422 - 19428. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
D. Lopez, T. W. Sandhoff, and M. P. McLean Steroidogenic Factor-1 Mediates Cyclic 3',5'-Adenosine Monophosphate Regulation of the High Density Lipoprotein Receptor Endocrinology, July 1, 1999; 140(7): 3034 - 3044. [Abstract] [Full Text]
|
|
|
|
|
|
C. Dorn, Q. Ou, J. Svaren, P. A. Crawford, and Y. Sadovsky Activation of Luteinizing Hormone beta Gene by Gonadotropin-releasing Hormone Requires the Synergy of Early Growth Response-1 and Steroidogenic Factor-1 J. Biol. Chem., May 14, 1999; 274(20): 13870 - 13876. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
A. J. Reinhart, S. C. Williams, B. J. Clark, and D. M. Stocco SF-1 (Steroidogenic Factor-1) and C/EBP{beta} (CCAAT/Enhancer Binding Protein-{beta}) Cooperate to Regulate the Murine StAR (Steroidogenic Acute Regulatory) Promoter Mol. Endocrinol., May 1, 1999; 13(5): 729 - 741. [Abstract] [Full Text]
|
|
|
|
|
|
P. De Santa Barbara, N. Bonneaud, B. Boizet, M. Desclozeaux, B. Moniot, P. Sudbeck, G. Scherer, F. Poulat, and P. Berta Direct Interaction of SRY-Related Protein SOX9 and Steroidogenic Factor 1 Regulates Transcription of the Human Anti-Mullerian Hormone Gene Mol. Cell. Biol., November 1, 1998; 18(11): 6653 - 6665. [Abstract] [Full Text]
|
|
|
|
|
|
A. L. Jacob and J. Lund Mutations in the Activation Function-2 Core Domain of Steroidogenic Factor-1 Dominantly Suppresses PKA-dependent Transactivation of the Bovine CYP17 Gene J. Biol. Chem., May 29, 1998; 273(22): 13391 - 13394. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
P. A. Crawford, C. Dorn, Y. Sadovsky, and J. Milbrandt Nuclear Receptor DAX-1 Recruits Nuclear Receptor Corepressor N-CoR to Steroidogenic Factor 1 Mol. Cell. Biol., May 1, 1998; 18(5): 2949 - 2956. [Abstract] [Full Text]
|
|
|
|
 |
|
 Y. Sadovsky and P. A. Crawford Developmental and Physiologic Roles of the Nuclear Receptor Steroidogenic Factor-I in the Reproductive System Reproductive Sciences, January 1, 1998; 5(1): 6 - 12. [Abstract] [PDF]
|
|
|
|
|
|
D. Liu, M. Chandy, S.-K. Lee, Y. Le Drean, H. Ando, F. Xiong, J. Woon Lee, and C. L. Hew A Zebrafish Ftz-F1 (Fushi Tarazu Factor 1) Homologue Requires Multiple Subdomains in the D and E Regions for Its Transcriptional Activity J. Biol. Chem., May 26, 2000; 275(22): 16758 - 16766. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
H. Pincas, J.-N. Laverriere, and R. Counis Pituitary Adenylate Cyclase-activating Polypeptide and Cyclic Adenosine 3',5'-Monophosphate Stimulate the Promoter Activity of the Rat Gonadotropin-releasing Hormone Receptor Gene via a Bipartite Response Element in Gonadotrope-derived Cells J. Biol. Chem., June 22, 2001; 276(26): 23562 - 23571. [Abstract] [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 |
