Minireview: Steroidogenic Factor 1: Its Roles in Differentiation, Development, and Disease
Bernard P. Schimmer and Perrin C. White
Banting and Best Department of Medical Research (B.P.S.), University of Toronto, Toronto, Ontario, Canada M5G 1L6; and Department of Pediatrics (P.C.W.), University of Texas Southwestern Medical Center, Dallas, Texas 75390-9063
The orphan nuclear receptor steroidogenic factor 1 (SF-1, also called Ad4BP, encoded by the NR5A1 gene) is an essential regulator of endocrine development and function. Initially identified as a tissue-specific transcriptional regulator of cytochrome P450 steroid hydroxylases, studies of both global and tissue-specific knockout mice have demonstrated that SF-1 is required for the development of the adrenal glands, gonads, and ventromedial hypothalamus and for the proper functioning of pituitary gonadotropes. Many genes are transcriptionally regulated by SF-1, and many proteins, in turn, interact with SF-1 and modulate its activity. Whereas mice with heterozy- gous mutations that disrupt SF-1 function have only subtle abnormalities, humans with heterozy- gous SF-1 mutations can present with XY sex reversal (i.e. testicular failure), ovarian failure, and occasionally adrenal insufficiency; dysregulation of SF-1 has been linked to diseases such as en- dometriosis and adrenocortical carcinoma. The current state of knowledge of this important transcription factor will be reviewed with a particular emphasis on the pioneering work on SF-1 by the late Keith Parker. (Molecular Endocrinology 24: 1322-1337, 2010)
T he orphan nuclear receptor, steroidogenic factor 1 (SF-1), plays a key role in the development and func- tion of steroidogenic tissues. It was discovered by Keith Parker, who died tragically at age 54 in 2008. Parker’s work established an extremely active area of investigation that has resulted in more than 1000 publications listed in PubMed. Our contributions to SF-1 biology (and connections to Parker) began in 1984 when we met at a symposium. White had recently cloned the cytochrome P450 (CYP)21 gene en- coding steroid 21-hydroxylase, which is mutated in most cases of congenital adrenal hyperplasia. Schimmer suggested a collaboration to explore the regulation of this gene, and steroidogenesis in general, using mouse Y1 adrenocortical cells, which lack Cyp21 expression. White referred him to another collaborator, Jon Seidman at Harvard, whose lab- oratory had isolated mouse cosmids carrying the Cyp21 gene. In turn, Seidman assigned the project to Keith Parker, a postdoctoral fellow in his laboratory at the time.
These studies demonstrated that Y1 cells stably trans- fected with the mouse Cyp21 cosmid recovered hormonally regulated expression of 21-hydroxylase (1). Sequences es- sential for expression were localized to the proximal 330 bp of 5’-flanking DNA (2, 3). When genes encoding other ste- roid hydroxylases were isolated and their 5’-flanking re- gions compared, shared AGGTCA promoter elements were identified in several of these genes that interacted with the same DNA-binding protein in steroidogenic cell lines. This protein was designated steroidogenic factor 1 (SF-1) or ad- renal 4-binding protein (Ad4BP) (see Ref. 4 for a review of these studies). The selective expression of SF-1 in steroido- genic cells and its regulation of genes encoding several dis- tinct steroid hydroxylases provided the first clues that it was a key determinant of cell-selective expression of steroido- genic enzymes.
Parker and colleagues (5) reasoned that the AGGTCA DNA recognition motif represented a binding site for an
Abbreviations: AHC, Adrenal hypoplasia congenital; AMH, anti-Müllerian hormone (Müllerian-inhibiting substance); CB1R, cannabinoid receptor 1; CDK, cyclin-dependent kinase; CNS, central nervous system; CYP, cytochrome P450; DAX-1, dosage-sensitive sex reversal-AHC critical region on the X chromosome gene 1; DBD, DNA-binding domain; @-GSU, glycoprotein hormone «-subunit; MC2R, melanocortin receptor 2 (corticotropin re- ceptor); SF-1, steroidogenic factor 1; SOX, SRY (sex determining region Y)-box; SRA, steroid receptor RNA activator; StAR, steroidogenic acute regulatory protein; SUMO, small ubiquitin- related modifier; USF, upstream stimulatory factor; VMH, ventromedial hypothalamus.
atypical member of the nuclear hormone receptor family and cloned mouse SF-1 cDNA from a Y1 cell cDNA li- brary using a hybridization probe comprising the DNA- binding region of retinoid X receptor. Independently, Morohashi and colleagues (6) used an oligonucleotide affinity column to purify the protein from bovine adrenal glands, ultimately allowing them to obtain amino acid sequence and clone a bovine cDNA with an oligonucleo- tide probe. Subsequent studies established that the mouse and bovine cDNAs encoded orthologs of a protein that transactivated the steroid hydroxylase promoters in ste- roidogenic and nonsteroidogenic cells. The sequences of these cDNAs confirmed that SF-1 belonged to the nuclear hormone receptor family, with striking homology to the Drosophila nuclear receptor fushi tarazu factor 1. SF-1 was also similar in sequence to an orphan receptor cloned from mouse liver, designated LRH1, and its human ho- molog, PHR-1. Although LRH1 clearly derives from a separate gene and has an expression profile that is distinct from that of SF-1 (e.g. in liver and pancreas), the LRH1 and SF-1 sequences are sufficiently similar to group them as members of the same subfamily of nuclear hormone receptors, designated NR5A. The SF-1 gene is now offi- cially termed NR5A1 (in this review, we will continue to use SF-1 to refer to the protein or mRNA) and the LRH1 gene is named NR5A2. Early work on SF-1 structure and function has been reviewed elsewhere (4).
Like other nuclear hormone receptors, SF-1 has a mod- ular domain structure comprised of an N-terminal zinc finger DNA-binding domain (DBD), a ligand-binding do- main, a C-terminal AF-2 activation domain, and an inter- vening proline-rich hinge region that has AF-1 like acti- vation activity (Fig. 1). SF-1 also contains a fushi tarazu factor 1 or A box (i.e. a 30-amino acid extension of the DBD) that mediates specific binding to sequences 5’ to the consensus hexamer. As determined by X-ray crystallogra- phy of the DBD complexed with a sequence in the proximal promoter region of the INHA (inhibin-a) gene, SF-1 forms a specific complex with DNA by binding to the major and minor grooves through the core DBD and the N-terminal segment of the A box, respectively. Additionally, a helix in the C-terminal segment of the A box interacts with both the core DBD and DNA and serves as an important determinant of stability of the complex. This helix may also interact with coactivators and other DNA-bound factors (7).
Regulation of SF-1 Expression and Function
Patterns of expression in adult and embryonic tissues
The localization of SF-1 in adult tissues has been in- vestigated extensively in mice, rats, humans, and other vertebrates. Consistent with its role as a master regulator
M1I,D6A1bp
E11X,V15M,C16X
C33S
ZF1
V41G →
DBD
+G35E
M78I
ZF2
R84H, R84C
G91S
+R92Q
Q107X
A box
G123S &P129L
P13041bp
Y138X, G146A
Pro
H
E225A1bp
L231A9bp
D273N →
< R255L
R2
E35348bp V355M =
LBD
R3
L437Q →
AF2
of steroidogenesis, SF-1 is expressed in the major steroi- dogenic tissues, i.e. the three zones of the adrenal cortex, testicular Leydig and Sertoli cells and ovarian intersti- tium, theca cells, granulosa cells, and, to a lesser degree, corpus luteum (6, 8-11). It also is expressed in pituitary gonadotrophs where it regulates the expression of the gonadotropins (12, 13) and the GnRH receptor (14). Apart from these tissues, SF-1 expression is limited to neurons in the dorsomedial portion of the ventromedial hypothalamus that are distinct from those expressing GnRH, to the endothelial linings of the venous sinuses and pulp veins in the spleen (11, 15), and to a subset of neurons in the hippocampus that coexpress steroidogenic acute regulatory protein (StAR) and aromatase (16).
In developing embryos, SF-1 first appears in the uro- genital ridge and represents the earliest marker of adrenal and gonadal differentiation (17-20). As development proceeds, SF-1 is expressed in the steroid-secreting corti- cal zones of the adrenal gland, in testicular Leydig and Sertoli cells, and in the human ovary; however, in mice and rats, ovarian expression of SF-1 is turned off as de- velopment proceeds and does not resume until just before
birth (17, 19, 20). SF-1 also appears in the fetal spleen (15) and in the prosencephalon-that region of the brain that includes the hypothalamic primordium (19).
Regulation of NR5A1 promoter activity
Regulatory elements and transcription factors
The first 110 bp in the 5’-flanking region of NR5A1 do not contain a TATA box but do include a small number of regulatory elements including a SRY (sex determining re- gion Y)-box (SOX) binding site, which may contribute to Sertoli cell expression of SF-1 (21), and an E box, a CCAAT box, and an Sp1/Sp3 site that may function more broadly (22-26)(Fig. 2). In vitro, these elements bind combinations of proteins that differ somewhat among cell lines of adrenal, Leydig, Sertoli, and pituitary origin (21- 23, 25-27). Two other Sp1/Sp3 sites situated downstream of the transcription start site between +10 and +30 also contribute to optimal activity of the promoter (25). Longer NR5A1 promoter fragments with 589 bp of 5’-
A
100 kb
+H
HE-HE
HH
H
NR6A1
NR5A1 GPR144
PSMB7
B
10 kb
1
2,3
4
5
6
7
Gonad
Fetal adrenal
VMH Pituitary
C
Sox9
USF
NFY
Sp
Sp
Sp
Sx
E
C
Sp
-100
-80
-60
-40
-20
0
+20
flanking sequence contain binding sites for GATA-4 (28) and for WT1 and Lhx9 (29) that enhance the activity of the promoter in testes-derived cells. Other transcription factors implicated in NR5A1 regulation include SOX15, SOX30, and TEAD-4 (21, 30). Finally, the Polycomb or- tholog, chromobox homolog 2 (CBX2, M33), may be an important regulator of NR5A1. CBX2 binds the NR5A1 promoter in Y1 cells. Mice carrying CBX2 null mutations have reduced SF-1 expression and have phenotypes rem- iniscent of those seen with SF-1 mutations (see below) including XY sex reversal, small adrenal glands, and ab- normalities in the spleen (31). XY sex reversal also is seen in humans with CBX2 mutations (32).
Studies of transgenic animals give a somewhat differ- ent picture of the DNA segments required for normal NR5A1 expression. The very short NR5A1 promoter fragments are insufficient to recapitulate gene expression in sites where SF-1 is normally expressed (33); the some- what larger fragment from -590 to +85 is able to direct gene expression in the indifferent gonads of transgenic mouse embryos in a WT1-dependent manner but is insuf- ficient to drive expression in any other SF-1-expressing tissue (29). A much larger reporter gene driven by mouse genomic DNA, extending from exon 2 into the upstream gene NR6A1 gene encoding germ cell nuclear factor, is able to drive gene expression in the adrenal cortex, testic- ular Leydig cells, ovarian theca cells, the ventromedial hypothalamus (VMH), and the spleen, but not in the pi- tuitary gland or corpus luteum (33). So far, only a rat genomic fragment including the NR5A1 gene as well as flanking DNA extending into the NR6A1 and PSMB7 genes on either side duplicates the patterns of endogenous SF-1 expression, both spatially and quantitatively (34). Within this very large genomic fragment lie highly con- served regions within intron 6 of NR5A1 that are re- quired to recapitulate VMH-specific (35) or gonado- trope-specific expression (36) of SF-1 from the fetal stage to the adult (Fig. 2).
Expression of the NR5A1 gene in the fetal adrenal gland, on the other hand, depends on binding sites for Pbx1-Prep and Pbx1-hox complexes and for SF-1 itself within a highly conserved region of intron 4 (37). This region is unable to sustain SF-1 expression in the adult gland and thus is considered to be a fetal adrenal-specific enhancer of NR5A1 expression that, by binding SF-1, functions in an autoregulatory manner. Consistent with this, mice carrying an SF-1 transgene that contains this region (along with 5.8 kb of 5’-flanking sequences) have increased adrenal size and ectopic adrenal tissue in the thorax, suggesting that increased levels of SF-1 may divert uncommitted precursors to the steroidogenic lineage (38). Moreover, all cells in the murine adult adrenal cortex
descend from cells that express SF-1 under the control of this enhancer before d 14.5 of embryogenesis (Fig. 2; Ref. 39).
Contributions of DNA methylation
Whereas transcription factor-binding sites and their interacting proteins are primary determinants of NR5A1 expression, DNA methylation appears to act as a second- ary layer of control. CpG islands in the NR5A1 promoter isolated from endometriotic tissue are hypomethylated when compared with those from normal endometrial stroma and are associated with the aberrant expression of NR5A1 and other genes involved in steroidogenesis (40). Treatment of SF-1-negative, normal endometrial stromal cells with the demethylating reagent, 5-aza-deoxycyti- dine, leads to demethylation of the NR5A1 promoter and the appearance of NR5A1 transcripts, whereas methyl- ation of the promoter leads to a loss of its activity. The correlation between methylation of the NR5A1 promoter and NR5A1 expression also is seen in steroid-secreting cell lines and normal steroid-secreting tissues (41). DNA methylation at CpG-rich sequences generally increases with cellular differentiation, providing an efficient mech- anism for transcriptional repression and maintenance of cell-specific phenotypes (42). Consistent with this general finding, the NR5A1 proximal promoter is unmethylated in the developing testis and ovary, whereas it is hyperm- ethylated in most embryonic tissues in which SF-1 is not expressed. Embryonic mouse and human kidneys are an exception because SF-1 is never expressed but, for un- known reasons, the NR5A1 gene is nevertheless hypom- ethylated (41). On balance, the data suggest that methyl- ation of the proximal promoter region of NR5A1 is established early in differentiation and contributes to the tissue-restricted expression of SF-1 during development.
Chromatin interactions
NR5A1 is located only 13 kb downstream of the last exon of the NR6A1gene (Fig. 2), which has a distinct expression pattern, suggesting the presence of an inter- vening insulator element (a DNA fragment between two genes that functions as an enhancer blocker and a barrier to modified chromatin). In Y-1 cells, there are three hy- persensitive sites between the last exon of NR6A1 and the first exon of NR5A1. The most upstream site contains a binding site for CCCTC-binding factor, a zinc finger protein known to bind insulator elements, whereas the other sites associate with the nuclear matrix (i.e. they constitute a ma- trix attachment region). As determined by chromatin immu- noprecipitation analysis, there is a discontinuous pattern of histone acetylation upstream of these sites, suggesting that the chromatin architecture specified by CCCTC-binding factor and the matrix contribute to the distinct pattern of transcriptional regulation of NR5A1 and NR6A1 (43).
Signal transduction pathways affecting SF-1 function
cAMP-dependent signaling
The effects of SF-1 on cell-selective gene expression are intimately associated with cAMP-regulated expression of many of the same genes, e.g. CYP11A1, CYP11B1, CYP11B2, CYP17, CYP21, STAR (44), SCARB1 (45, 46), MC2R (47, 48), and INHA (49). This interplay be- tween SF-1 and the cAMP/protein kinase A-signaling cas- cade is complex and involves multiple factors acting in promoter-specific contexts that have not been fully de- fined. Although SF-1 contains a consensus site for phos- phorylation by protein kinase A (6), mutating this does not affect the cAMP-dependent regulation of its activity, so it is unlikely that this synergy results from protein kinase A-mediated phosphorylation of SF-1 (50, 51). One facet of cAMP-dependent signaling may be the generation of activating ligands (see below) for SF-1 via activation of diacylglycerol kinase § and the resultant synthesis of phosphatidic acid (52, 53). The cAMP-signaling cascade also appears to affect the recruitment of SF-1 to active foci of transcription within the nucleus and the dynamic interaction of SF-1 with its regulatory cofactors (48, 54, 55). This effect may be secondary to ligand activation of SF-1 (52) or to cAMP-mediated induction of p300 (54). Finally, cAMP may affect the levels of SF-1 RNA and protein; however, evidence supporting this hypothesis is conflicting (56-59).
MAPK and cyclin-dependent kinase 7 (CDK7)
SF-1 is phosphorylated primarily at S203, within the activation factor helix 1 domain, the hinge region be- tween the DBD and ligand-binding domain of the protein (60, 61). The MAPK, Erk2, can phosphorylate SF-1 at S203 in vitro and possibly in vivo (60, 62) and has been implicated in the estrogen-dependent activation of SF-1 via the G protein-coupled receptor, GPR30 (63). CDK7 also may contribute significantly to the phosphorylation of SF-1 at S203 (64). Both the ligand-binding domain of SF-1 and the extent of posttranslational modification via small ubiquitin-related modifier (SUMO)-ylation (see be- low) modify the extent of SF-1 phosphorylation at S203. Phosphorylation of SF-1 at S203 does not affect its half- life, its subcellular localization, or its binding to DNA (60); however, it does enhance its ability to interact with cofactors such as glucocorticoid receptor-interacting protein 1 (GRIP1) and silencing mediator of retinoid and thyroid hormone receptor (SMRT) (60), to sustain SF-1-dependent activity at some promoters but not oth- ers (51, 64), and to adopt an active conformation within its ligand-binding domain (61).
Protein kinase C-8
SF-1 expression may be under the control of a regulatory module comprised of erb-b3, ebp1, and protein kinase C-8, because the knockdown of these three transcripts using spe- cific small interfering RNAs in a heterologous cell culture system enhances SF-1 promoter activity (30); conversely, overexpression of protein kinase C-8 inhibits SF-1 promoter activity. The extent to which the regulatory activities of pro- tein kinase C-8, erb-b3, and ebp1 at the SF-1 promoter can be extrapolated to SF-1 expression in situ in cells that nor- mally express SF-1 has yet to be determined.
Wnt and hedgehog
The Wnt signaling pathway also impacts on SF-1- regulated gene expression. This pathway is comprised of Wnt ligands, the Frizzled family of G protein-cou- pled receptors, low-density lipoprotein receptor-re-
lated protein 5/6 coreceptors, Dishevelled, ß-catenin, and a ß-catenin inactivation complex that keeps ß-cate- nin at low levels and localized to the plasma membrane (65). The activation of the Wnt pathway leads to phos- phorylation of Dishevelled, the consequent inhibition of the ß-catenin inactivation complex, and accumulation of ß-catenin in the nucleus. ß-Catenin binds directly to SF-1 and synergistically regulates the expression of Anti-Mülle- rian hormone (AMH; Mullerian-inhibiting substance) re- ceptor 2 (AMHR2) a-inhibin, DAX-1 (dosage-sensitive sex reversal-AHC critical region on the X chromosome gene 1), and several genes involved in steroidogenesis, including STAR and CYP19A1 (see Table 1). Targeted inactivation of ß-catenin in SF-1-expressing cells using an SF-1/Cre transgene leads to adrenal agenesis, supporting the idea that proper functioning of the Wnt/B-catenin pathway is critical for normal adrenal development (66).
| Site of action | Target genes | Comments |
|---|---|---|
| Ventromedial hypothalamus | NMDAR1 | N-methyl-D-aspartate receptor |
| BDNF | Brain-derived neurotrophic factor | |
| A2BP1, AMIGO2, CDH4, NPTX2, | Cell adhesion and cell guidance molecules | |
| SEMA3A, SLIT3, NETRIN3 | ||
| FEZF1, NKX2-2 | Transcription factors | |
| Gonadotropes | CGA | @-Subunit of glycoprotein hormones |
| LHB | Luteinizing hormone (LH) ß | |
| FSHB | Follicle-stimulating hormone (FSH) ß | |
| GNRHR | Gonadotropin-releasing hormone receptor | |
| Adrenal cortex Gonads | CYP11A1, CYP17, CYP21, | Cytochrome P450 steroid hydroxylases |
| CYP11B1, CYP11B2 | ||
| HSD3B2 | 3ß-Hydroxysteroid dehydrogenase; Hsd3b1 in rodents | |
| STAR | Steroidogenic acute regulatory protein | |
| MC2R | Corticotropin receptor | |
| SCARB1 | Scavenger receptor-B1 | |
| NR0B1 | Dosage-sensitive sex reversal, adrenal | |
| hypoplasia congenital, X chromosome | ||
| (DAX-1) | ||
| AKR1B7 | Aldose reductase-like protein | |
| SULT2A1 | Steroid sulfotransferase 2A1 | |
| ADCY4 | Adenylyl cyclase type 4 (repressor activity) | |
| Leydig cells | CYP11A1, CYP17 | Cytochrome P450 steroid hydroxylases |
| STAR | Steroidogenic acute regulatory protein | |
| LHR | LH receptor | |
| INSL3 | Insulin-like 3 | |
| PRLR | Prolactin receptor | |
| AMHR2 | Anti-Mullerian hormone/Mullerian | |
| inhibiting substance receptor | ||
| Sertoli cells | AMH | Anti-Mullerian hormone/Mullerian inhibiting substance |
| INHA | Inhibin &-subunit | |
| FSHR | FSH receptor | |
| SRY | Sex-determining region Y (SRY) | |
| SOX9 | SOX9 (SRY box 9) | |
| Theca and granulosa cells | CYP11A1, CYP17, CYP19 | Cytochrome P450 steroid hydroxylases |
| STAR | Steroidogenic acute regulatory protein | |
| INHA | Inhibin & subunit | |
| OXT | Oxytocin |
The hedgehog pathway consists of three signaling li- gands: sonic hedgehog, desert hedgehog (the main signal- ing ligand in the male gonad), and Indian hedgehog. Each of these ligands interacts with its membrane receptor Patch to free Smoothen, a member of the G protein-cou- pled receptor family, for downstream effects that include inhibition of cleavage of the GLI3 transcription factor. GLI3 acts as a transcriptional activator, whereas its cleav- age product represses the expression of target genes (67, 68). The hedgehog pathway appears to have a role in SF-1 expression because disruption of the pathway decreases the gonadal expression of SF-1 (69), whereas activation of the pathway has the opposite effect (70).
Regulation of function through protein-protein and protein-RNA interactions
Although SF-1 interacts with its target DNA element as a monomer (71), its activity is strongly influenced by its interactions with other transcription factors and coregu- lators (for brief review see Ref. 72) (Table 2). Some inter- acting proteins are expressed in a tissue-specific manner whereas others are more widely expressed, and it is un- clear how each contributes to the coordinate and cell- selective expression of SF-1-dependent genes. Analyses of transcription complex assembly on the MC2R (48) and CYP17 (73) promoters indicate that SF-1-containing complex formation is dynamic and cyclical. Thus the na- ture of the complexes formed is likely to be influenced by the relative abundance of the interacting partners and by their relative affinities for the promoter and for SF-1 at any given spatiotemporal interval.
Interactions between SF-1 and DAX-1 deserve particular comment (early work is reviewed in Ref. 74). Mutations in DAX-1 (NR0B1) cause X-linked adrenal hypoplasia con- genita (AHC) and hypogonadotrophic hypogonadism, phe- notypes similar to those seen with SF-1 mutations (see be- low). In early stages of mouse embryonic development, SF-1 expression in the urogenital ridge and brain either precedes or coincides with DAX-1 expression. As embryonic devel- opment proceeds, the two transcription factors exhibit sim- ilar expression patterns in the adrenal cortex, testis, ovary, hypothalamus, and anterior pituitary. The slightly earlier onset of SF-1 expression and its ability to bind specifically to a conserved sequence in the NROB1 5’-flanking region sug- gest that SF-1 may activate DAX-1 expression, and indeed SF-1 stimulates expression of the NR0B1 promoter in tran- sient expression assays in SF-1-deficient cells (75, 76).
Coexpression of DAX-1 and SF-1 inhibits SF-1-medi- ated transactivation of many target genes. DAX-1 inter- acts directly with SF-1 in vitro via the carboxy-terminal regions of each protein (77). Additionally, DAX-1 inter- feres with interactions between SF-1 and WT1 (the
| Protein | Gene(s) affected | Effect | Reference |
|---|---|---|---|
| Transcription factors | |||
| AR | LHB | - | 139 |
| c-JUN | CYP11A1 | + | 140 |
| DAX1 | AMH | - | 76, 78, 141 |
| EGR1 | LHB | + | 42 |
| FOXL2 | CYP19A1 | + | 143 |
| GATA4 | AMH | + | 144 |
| GR | DAX1 | + | 79 |
| NFKB | AMH | - | 145 |
| NFYA | FSHB | + | 117 |
| PTX1 | LHB | + | 146 |
| SOX8 | AMH | + | 147 |
| SOX9 | AMH | + | 148 |
| Sp1 | CYP11A1 | + | 149, 150 |
| WT1 | AMH | + | 141 |
| Coactivators | |||
| CBP/p300 | CYP11A1 | + | 151 |
| PCAF | CYP17 | + | 85 |
| GRIP1 | CYP21 | + | 60 |
| MBF1 | CYP11B1 | + | 152 |
| p/CIP | CYP17 | + | 153 |
| PNRC | CYP19 | + | 154 |
| SNURF | LHB | + | 155) |
| SRC1 | SF-1 binding site | + | 156, 157 |
| TIF2 | CYP17 | + | 153 |
| TReP-132 | CYP11A1 | + | 158 |
| Corepressors | |||
| CtBP1 | CYP17 | - | 73, 159 |
| DP103 | CYP11A1 | - | 160 |
| EID1 | CYP21 | - | 161 |
| GIOT1 | CYP21 | - | 162 |
| RIP140 | STAR, CYP17 | - | 163, 164 |
| SMRT | CYP21 | - | 60 |
| Zip67 | CYP11A1, CYP17 | - | 165 |
| Other proteins | |||
| B-Catenin | AMHR2, INHA, DAX1, STAR, | + | 166-170 |
| CYP19A1 | |||
| P54nrb/NonO | CYP17 | - | 171 |
| PSF | CYP17 | - | 171 |
| PIAS1, PIAS3 | SF-1 binding site | 172 |
Wilm’s tumor suppressor protein) or GATA4, each of which normally associates and synergizes with SF-1 to promote AMH expression. Finally, DAX-1 acts as an adaptor that recruits other factors, such as the nuclear receptor corepressor, to SF-1 (78).
Expression of DAX-1 in adrenal cells is decreased by ACTH and increased by glucocorticoid treatment, the latter occurring through a glucocorticoid receptor-SF-1 complex on the NR0B1 promoter. ACTH disrupts the formation of this complex by shifting SF-1 binding away from the NR0B1 promoter and toward the MC2R and STAR promoters (79, 80).
Whereas all of these findings suggest that DAX-1 acts mainly to negatively regulate SF-1 actions (and thus ste- roidogenesis), the inhibitory domain in DAX-1 is deleted
or mutated in most AHC mutants, suggesting that loss of the inhibitory property in DAX-1 is associated with the development of AHC. Given that inactivating mutations in DAX-1 and SF-1 have similar phenotypes, this seems paradoxical and possibly reflects DAX-1 functions in de- velopment that are distinct from its regulatory roles in postnatal life. Indeed, DAX-1 can function as an SF-1 coactivator in some contexts. Both SF-1 and DAX-1 bind to steroid receptor RNA activator (SRA), a noncoding RNA that functions as a coactivator. In cotransfection experiments, an SRA expression plasmid increases the ability of SF-1 (or DAX-1) plasmids to transactivate re- porter constructs, whereas knockdown of SRA expres- sion abolishes coactivation by Dax-1. Moreover, SRA is physically associated with both SF-1 and DAX-1 in Y1 cells and is expressed in both the adrenal glands and go- nads. The coactivator TIF2 also associates with DAX-1 and synergistically coactivates SF-1 target gene transcrip- tion. Corepressor vs. coactivator functions of DAX-1 may be dose dependent; although transfection of DAX-1 expression constructs usually inhibits expression of ste- roidogenic proteins, knockdown of endogenous DAX-1 actually down-regulates the expression of CYP11A1 and STAR in both H295R adrenal and MA-10 Leydig cells (81).
Regulation through posttranslational modification
In addition to phosphorylation at S203 as describe above, SF-1 is subject to SUMO conjugation and acetyla- tion at 8-amino groups of target lysine residues. SUMO- ylation represses SF-1 function (82-84), whereas acetyla- tion enhances its activity (54, 85).
For a brief review of protein SUMO-ylation and its consequences on gene expression, see Ref. 86. SF-1 is SUMO-ylated within its DBD at K119 and within its hinge region at K194 (82-84); K194 appears to be the primary site of SUMO-ylation and suppression of SF-1 activity whereas K119 makes a minor contribution (82-84). Dis- ruption of these SUMO-ylation sites increases the promoter- regulatory activity of SF-1 at CYP19, HSD 3B2, STAR, and at a number of isolated SF-1 response elements (82, 84); preventing the SUMO-ylation of SF-1 by overex- pressing the SUMO-isopeptidase, SENP1, also enhances SF-1 activity (84). SUMO-ylation does not affect the sta- bility, cellular localization, or dynamic association of SF-1 with chromatin or its ability to bind to canonical SF-1 response elements but does inhibit its phosphoryla- tion by CDK7 at S203 (87, 88) and may (84) or may not (82) affect its ability to interact with repressors such as DP10; SUMO-ylation at the minor K119 site interferes with the ability of SF-1 to bind to and function at nonca- nonical response elements such as those found in the rat INHA promoter (87).
Two histone acetyltransferase proteins, PCAF (85) and p300, (54) stimulate the incorporation of labeled acetate into SF-1 and increase its transcriptional activity. Disrup- tion of putative acetylation sites prevents acetylation of SF-1 and decreases its transcriptional activity (54, 85), whereas inhibition of the deacetylation of SF-1 using tri- chostatin A increases both its half-life and activity (85, 89). The acetylation of SF-1 by p300, but not by PCAF, appears to increase its localization within transcription- ally active nuclear foci (54). Other coactivators with his- tone acetyltransferase activity interact with, and modu- late, the activity of SF-1 as well (Table 2).
The dynamics of SUMO-ylation and acetylation are incompletely understood. Less than 10% of SF-1 appears to be SUMO-ylated in vivo (82-84, 88), and it is not clear how this small fraction can have a dominant effect on SF-1-dependent transcription. The same can be said for acetylated SF-1, based on the presumption that it too represents only a small fraction of the total. Perhaps the posttranslational modifications of SF-1 are influenced by the cyclical and dynamic recruitment of SF-1 into active transcription complexes because, for example, DNA- bound SF-1 is refractory to SUMO-ylation at K113 (87).
Phospholipid-dependent regulation
The finding that SF-1 is a member of the nuclear receptor family of transcription factors engendered a search for li- gands that regulate its function. Oxysterols enhance SF-1 function in Chinese hamster CV-1 lung cells (90, 91) but not in other cell types (61, 91, 92). X-ray crystallography and mass spectrophotometry of bacterially expressed SF-1 indi- cate that its ligand-binding pocket is very large, very hydro- phobic, and occupied by phosphatidylethanolamine or phosphatidylglycerol (93-96). These bacterial phospholip- ids can be exchanged for other potential regulatory ligands such as phosphatidic acid, phosphatidylcholine, and phos- phatidylinositol di- and triphosphates (93-95) with little consequence on the conformation of the AF2-activating do- main of SF-1, but with some effect on conformation at the entrance to the ligand-binding pocket of the protein (95). SF-1 purified from H295R adrenal tumor cells is associated with sphingosine, which acts as an inhibitor of SF-1 function in reporter gene assays (53). Treatment of cells with cAMP increases sphingosine catabolismdecreases the amount of sphingosine bound to SF-1, and enhances SF-1-dependent gene expression, whereas inhibitors of sphingosine synthesis enhance SF-1 activity. Sphingosine can be displaced from SF-1 to varying degrees by the phospholipid ligands dis- cussed above, suggesting that SF-1 function is controlled by changes in the lipid environment secondary to regulated changes in lipid metabolism. This may have a teleological explanation given the key role of SF-1 in regulating steroi-
dogenesis, a process requiring high concentrations of cholesterol.
As regards pharmacological ligands, low-molecular weight compounds with a rigid cis-bicyclo[3.3.p]oct-2-ene core structure selectively increase SF-1 activity (97) whereas 4-alkyloxy-phenol derivatives act as inverse agonists, sup- pressing its constitutive activity (98). Given the importance of SF-1 in steroidogenesis and in appetite control, these com- pounds may lead to the development of drugs useful in the treatment of steroid hormone excess, steroid hormone-de- pendent tumors, obesity, and related metabolic disorders.
The Role of SF-1 as a Regulator of Gene Expression-Lessons from In Vitro Studies
Gene targets of SF-1 in adrenal glands and gonads
Many SF-1 target genes (Table 1) have been identified with use of transient transfection assays using relatively limited stretches of promoter/regulatory DNA.
In adrenocortical cells, SF-1 increases expression of the corticotropin receptor (MCR2), STAR, and all of the en- zymes required for cortisol or corticosterone biosynthe- sis. It also increases levels of scavenger receptor B1, which is required for cellular importation of high-density li- poprotein cholesterol, the main source of cholesterol for steroidogenesis, and AKR1B7 (aldose reductase-like pro- tein), which metabolizes the isocaproaldehyde generated by cleavage of the cholesterol side chain (99). Although SF-1 is generally regarded as an activator of gene expres- sion, it is a strong negative regulator of the type 4 adenylyl cyclase, thereby regulating adenylyl cyclase isoform com- position (100) and of CYP11B2 (aldosterone synthase), which catalyzes the final step in aldosterone biosynthesis (101), thereby contributing to the restriction of aldoste- rone biosynthesis to the adrenal zona glomerulosa. In the ACTH-responsive, Y1 mouse adrenocortical tumor cell line, cells with a mutation affecting SF-1 function exhibit decreased expression of the MC2R CYP11B1 (11ß-hy- droxylase) CYP11A1 (cholesterol side-chain cleavage en- zyme), and STAR (102). In this cell line, the SF-1 defect affects the expression of the MC2R and CYP11B1 to a much greater degree than CYP11A1 or STAR, suggesting heterogeneity among these target genes in their regulation by SF-1.
SF-1 increases the expression of many testicular genes required to develop and maintain the male phenotype. Analogous to its effects in adrenocortical cells, SF-1 stim- ulates expression in Leydig cells of the LH receptor, STAR, and the CYP11A1 and CYP17 enzymes required for testosterone biosynthesis. Additionally, it increases expression of insulin-like polypeptide 3, which regulates testicular descent and acts in adults as a male germ cell
survival factor (103, 104), and the AMHR2 receptor for AMH, which is required for normal male reproductive tract development and serves as a negative regulator of Leydig cell steroidogenesis (105).
In Sertoli cells, SF-1 is required for the expression of the testes-determining gene products, sex-determining re- gion Y, and SOX9 AMH, the receptor for FSH, and INHA. Thus SF-1 is necessary for male sexual differenti- ation, a conclusion underscored by the XY sex reversal seen in either humans or mice carrying SF-1 mutations (see below).
Whereas SF-1 is required for ovarian development (as determined from studies of knockout mice, described later), interpreting the role of SF-1 in the adult ovary is complicated by the cyclic nature of follicular maturation, steroidogenesis, and corpus luteum formation. SF-1 is ex- pressed most highly in theca and interstitium, at lower levels in granulosa cells and at even lower levels in the corpus luteum. In contrast, LRH-1 (NR5A2) expression is abundant in cells involved in estrogen biosynthesis, granulosa cells during the estrous cycle (in rodents), and in corpora luteum in both rodents (106) and humans (107). LRH-1 potently transactivates both CYP19 (106) and hydroxysteroid dehydrogenase (107). Thus, LHR-1 may have a more prominent role than SF-1 in regulating luteal steroidogenesis, e.g. during pregnancy.
Strong evidence for a developmental role of SF-1 comes from studies of forced SF-1 expression in several different SF-1-negative cell types in primary culture. In the first of these studies, embryonic stem cells were in- duced to express the cholesterol side-chain cleavage en- zyme after the forced expression of SF-1 (108). Subse- quently, mesenchymal cells from mouse and human bone marrow cells were transformed into cells that expressed most of the enzymes required for steroid biosynthesis af- ter the forced expression of SF-1. These cells synthesized steroid hormones de novo, including progesterone, corti- costerone, cortisol, dehydroepiandrosterone, testoster- one, and estradiol in response to ACTH. In the human cells, SF-1 dramatically induced the expression of the recep- tors for both ACTH and LH (109, 110). Similar results were obtained using adipose tissue-derived mesenchymal cells, but in these cells, adrenal steroid biosynthesis (e.g. cortico- sterone) predominated, whereas gonadal steroids (e.g. tes- tosterone) were more apparent in bone marrow-derived cells (111). These results suggest the potential utility of adi- pose tissue-derived mesenchymal cells for autologous cell regeneration therapy for patients with adrenal insufficiency.
Although it was anticipated that SF-1 knockout mice might provide in vivo evidence for the importance of SF-1 in gene expression, particularly for those genes involved in steroidogenesis, the failure of the knockout mice to
develop the steroidogenic organs and the ventromedial hypothalamus (see below) precluded such analyses in these tissues. However, supporting in vivo roles of SF-1 have been provided for several genes listed in Table 1 including AMH (112, 113) and LHB (114).
Gene targets of SF-1 in pituitary and hypothalamus
The finding of SF-1 expression in pituitary gonado- tropes and in the VMH (see above) prompted a search for SF-1-regulated genes in these tissues. A series of in vitro DNA-binding assays and reporter gene assays of pro- moter activity suggested that the expression of the a-sub- unit of glycoprotein hormones [a-GSU (12)], LH ß (115), the GnRH receptor (116), FSH ß (117) in pituitary gona- dotropes were all regulated by SF-1 As discussed below, SF-1 clearly plays an important role in the expression of these genes in vivo as well.
In silico analyses of putative SF-1-binding sites among VMH-enriched transcripts, followed by promoter activity assays in vitro, suggested roles for SF-1 in the expression of the N-methyl-D-aspartate receptor, the cell adhesion mole- cules Amigo2, Cdh4, Sema3a, Slit3, and Netrin3, and other genes within the VMH, including Fezf1, Nptx2, Nkx2-2, and A2bp1 (118). It remains to be determined whether the SF-1-dependent expression of these transcripts can be con- firmed in the VMH-specific SF-1 knockout mice discussed below.
SF-1 as a Regulator of Differentiation and Development-Lessons from Knockout Mouse Models
Total disruption of SF-1
Consistent with the hypothesis that SF-1 is required for adrenal and gonadal steroidogenesis, SF-1 knockout mice die shortly after birth from adrenocortical insufficiency and exhibit male-to-female sex reversal of the external genitalia. However, these abnormalities are not due to poor expression of steroidogenic enzymes, but instead to complete absence of the adrenal glands and gonads (119, 120). The initial stages of adrenal and gonadal develop- ment occur in the absence of SF-1 but subsequently re- gress. Because their gonads regress before male sexual differentiation normally occurs, the internal and external urogenital tracts of SF-1 knockout mice are female, irre- spective of genetic sex. Heterozygous SF-1 knockout mice have decreased adrenal volume associated with impaired corticosterone production in response to stress (121).
The gonadotropes of SF-1 knockout mice also have impaired expression of a number of gene products that regulate reproduction, including LH-ß, FSH-ß, «GSU,
and the receptor for GnRH (13, 122). Moreover, knock- out mice lack the VMH, a hypothalamic region linked to feeding and appetite regulation and female reproductive behavior (122, 123). Finally, although the functional con- sequences remain to be defined, the SF-1 knockout mice have defects in their splenic parenchyma (15).
Tissue-specific knockout of SF-1
Anterior pituitary gland
Because mice with SF-1 completely disrupted are glo- bally deficient in SF-1, they die in the immediate postnatal period without corticosteroid replacement and cannot be used to delineate the roles of SF-1 at specific sites of ex- pression. Therefore the Cre/loxP system has been used to produce tissue-specific knockouts of SF-1. With use of a transgenic mouse line in which Cre recombinase expres- sion is directed to the anterior pituitary gland by the 5’- flanking sequences of the a-subunit of glycoprotein hor- mones, mice with pituitary-specific disruption of SF-1 have been generated. These @GSU-Cre/loxP mice selec- tively lack SF-1 immunoreactivity in the anterior pituitary (124) but have normal levels at other sites, including the adrenal cortex and VMH. These mice have markedly di- minished levels of pituitary gonadotropins and exhibit severe hypoplasia of the gonads and external genitalia. In males, the testes exhibit some signs of differentiation and development; however, germ cell numbers are consider- ably lower than normal and fail to progress to mature spermatids. Leydig cells are markedly reduced in number and are devoid of the histological features of steroidogen- esis. In females, the ovaries develop through the antral stage but do not produce large preovulatory follicles or corpora lutea. These results confirm earlier promoter- driven reporter gene studies indicating that SF-1 was re- quired for the expression of the gonadotropins and the GnRH receptor (reviewed in Ref. 4) and demonstrate that the local production of SF-1 in mice is essential for normal pituitary gonadotrope function.
Gonads
An anti-Müllerian hormone type 2 receptor-driven Cre recombinase transgene was used to generate mice with SF-1 specifically disrupted in testicular Leydig cells and ovarian granulosa cells (125). In the SF1-disrupted males, the testes failed to descend and were structurally abnor- mal and hypoplastic. Two hallmarks of Leydig cell func- tion, CYP11A and STAR expression, were impaired, in- dicating a defect in androgen biosynthesis. In females, ovaries were hypoplastic; adults were sterile and the ova- ries had reduced numbers of oocytes and lacked corpora lutea (126). These observations thus provide compelling
evidence for the essential role of SF-1 in normal reproduc- tive function.
Central nervous system (CNS)
To assess functions of SF-1 within the VMH, SF-1 was ablated within the CNS (the VMH being the sole site of SF-1 expression within the CNS) using a nestin-Cre trans- gene (127). Adrenals, gonads, and pituitaries all devel- oped and functioned normally in these mice, but the VMH nuclei were disrupted.
The VMH may be part of a medial hypothalamic de- fensive system mediating responses to predators (128). Consistent with this idea, these VMH-disrupted mice have increased anxiety-like behavior using many different tests, and their mediobasal hypothalamichave decreased expression or altered distribution of genes implicated in anxiety-like behavior including those encoding brain-de- rived neurotrophic factor, the type 2 CRH receptor (CRHR2), and urocortin 3. The BDNF (129) and CRHR2 genes both contain known SF-1-binding sites, suggesting that both are direct transcriptional targets of SF-1.
The VMH is also important for CNS regulation of appetite and energy homeostasis (see Ref. 130 for review). VMH neurons are implicated in satiety sensing, increas- ing excitatory input to proopiomelanocortin neurons in the arcuate nucleus and thereby activating anorexigenic neural circuits. In particular, SF-1-expressing neurons in the VMH make essential contributions to energy ho- meostasis as regulated by leptin and glucose signals. The cannabinoid receptor 1 (CB1R), which is associated with hypothalamic regulation of energy homeostasis, is highly expressed and functional in SF-1 neurons of the VMH and requires SF-1 for this expression. CB1R agonists ad- ministered systemically normally increase nocturnal food consumption in a partial satiety state but have blunted activities in CNS-specific SF-1 knockout mice. Conversely, the anorexic effect of a CB1R antagonist is significantly di- minished in these mice, arguing that regulatory roles of SF-1 on CB1R expression in the VMH play an important role in cannabinoid-induced effects on energy intake.
SF-1 and Human Disease
Mutations of SF-1
In contrast to homozygous SF-1 null mice, which can be kept alive with glucocorticoid replacement, no human has been identified who is homozygous for null mutations in the NR5A1 gene encoding SF-1. The first two SF-1- defective patients identified were heterozygous for muta- tions that abolished the ability of SF-1 to bind DNA, thus rendering the protein transcriptionally inactive without creating a dominant-negative effect. Both affected pa-
tients had adrenal insufficiency; one was a phenotypic female XY infant, and the other was an apparently nor- mal XX female who was not diagnosed with adrenal in- sufficiency until 14 months of age (131, 132). A third infant, who had adrenal failure and complete 46,XY sex reversal, was homozygous for a mutation that altered a highly conserved residue in a secondary DBD that causes partial loss of DNA binding and transcriptional activity (133). Heterozygous carriers of this mutation were nor- mal; thus, heterozygosity for apparently null mutations or homozygosity for a mutation that only partially reduces activity yield similar phenotypes.
These data suggested that, in humans, male gonad de- velopment and adrenal development in both sexes re- quired SF-1 expression in a dosage-sensitive manner. However, heterozygous SF-1 mutations subsequently were identified mainly among undervirilized 46,XY indi- viduals (i.e. XY disorders of sexual development) who do not have adrenal insufficiency (134). Thus, the SF-1 ge- notype alone seems insufficient to explain why some pa- tients have adrenal insufficiency, whereas others do not.
Additionally, SF-1 mutations impairing transcrip- tional activity have been identified among 46,XX women with premature ovarian failure in four kindreds with his- tories of both 46,XY disorders of sex development and 46,XX primary ovarian insufficiency and in two of 25 subjects with sporadic ovarian insufficiency. None of these subjects had clinically apparent adrenal insuffi- ciency. A range of ovarian anomalies was found including 46,XX gonadal dysgenesis and primary ovarian insuffi- ciency (135).
The adverse effects of heterozygous SF-1 mutations on reproductive function in both males and females could explain the failure to identify humans who are homozy- gous for null mutations, although it remains possible that such a condition is embryonically lethal in humans, per- haps (for example) owing to adverse effects on CNS development.
Dysregulation of SF-1
Endometriosis
SF-1 may play a significant role in the pathogenesis of endometriosis; it is undetectable in normal endometrial stromal cells but is expressed in endometriotic cells. This seems to be due, in part, to hypomethylation of a CpG- rich region in the proximal promoter region of the gene (see above). Expression of SF-1 leads to aberrant expres- sion of the STAR and CYP19 genes, which, in turn, in- crease the endogenous synthesis of estrogen within endo- metriotic tissue, a key causative factor in the disease. Additionally, the E-box sequence (CACGTG) at -82/ -77 in the SF-1 promoter is required for its activity in
endometriotic stromal cells as in most other cell types. This element interacts with upstream stimulatory fac- tors (USF) 1 and 2; USF2 mRNA and immunoreactive USF2 levels are increased in endometriotic tissues com- pared with normal endometrium, and knockdown of USF2 abolishes the overexpression of SF-1 (for review see Ref. 136).
Adrenocortical carcinoma
Because deficiency of SF-1 causes adrenal agenesis, it would not be surprising if SF-1 overexpression was asso- ciated, conversely, with adrenocortical overgrowth. In a cohort of children with adrenocortical carcinoma from Brazil (where this condition occurs relatively frequently), the 9q33.3 chromosomal region carrying the NR5A1 gene is amplified in almost all cases, and NR5A1 gene copy number is increased (137). SF-1 protein is overex- pressed as well, but levels of expression do not correlate with gene copy number or with clinical grade (138). The latter fact implies that SF-1 overexpression is per- missive for tumorigenesis but not related to malignant progression.
Conclusions
Since its discovery in 1992, much has been learned about the roles of SF-1 in gene expression, differentiation and development, and disease. Nevertheless, many of the questions that were raised 5 yr after its discovery (4) re- main unresolved today. Some of these are touched on below along with new questions raised as the result of recent progress.
What mechanisms regulate the expression of SF-1?
Whereas specific genetic regions have been identified as necessary for SF-1 expression in the gonads, fetal adrenal gland, pituitary gonadotropes, and VMH (see above), the regions required for SF-1 expression in the adult adrenal cortex are as yet unknown, as are many of the transacting factors that presumably regulate the or- dered expression of SF-1 in the various tissues.
What mechanisms regulate SF-1 function?
SF-1 clearly is a master regulator of transcription that directs cell-selective gene expression within the endocrine system; however, its activity is regulated by phospholip- ids, by a large array of transcription factors and coregu- lators, and by posttranscriptional modification. Whether phospholipids ligands serve as SF-1 activators or merely as stabilizers of SF-1 function is uncertain and although a regulatory role for sphingosine as an inhibitory ligand is somewhat more compelling, it remains to be determined whether its actions can be extended beyond a simple re-
porter gene assay in a specific cell line. Furthermore, the fact that SF-1 interacts with such a large number of pro- teins that modify its activity in vitro (Table 2) suggests a promiscuity that seems inconsistent with the tissue-spe- cific functions of SF-1. Ultimately, these interactions must be placed in a context that explains the temporal as well as the tissue-specific and gene-specific effects of SF-1.
What contributions does SF-1 make to appetite control and anxiety?
The VMH-specific knockout of SF-1 in mice leads to appetite and anxiety disorders; however, it is not clear whether SF-1 function in this context is merely permissive and related to the development of the VMH, or whether SF-1 regulates the expression of genes in the VMH that directly contribute to the aberrant phenotypes.
How do SF-1 mutations in humans impact on phenotype?
The apparent discordance in individual phenotypes produced by SF-1 mutations that disrupt SF-1 function requires further investigation. Although the SF-1 muta- tions described to date have been found in patients with adrenal and/or gonadal deficiency, one wonders if new mutations will be found with phenotypes resulting from effects at other sites, e.g. the VMH or spleen.
Acknowledgments
We respectfully dedicate this paper to our late friend and col- league, Keith Parker.
Address all correspondence and requests for reprints to: Dr. Perrin C. White, Department of Pediatrics, University of Texas Southwestern Medical Center, 5323 Harry Hines Bou- levard, Dallas, Texas 75390-9063. E-mail: perrin.white@ utsouthwestern.edu.
This work was supported by Grant MOP-64325 from the Canadian Institutes of Health Research (to B.P.S.) P.C.W. is the Audry Newman Rapoport Distinguished Chair in Pediatric Endocrinology.
Disclosure Summary: P.C.W. has nothing to disclose. B.P.S. receives royalties from McGraw-Hill Inc. for his contributions as an author of Goodman and Gilman’s The Pharmacological Basis of Therapeutics (11th ed. New York; McGraw-Hill, Inc.).
References
1. Parker KL, Chaplin DD, Wong M, Seidman JG, Smith JA, Schimmer BP 1985 Expression of murine 21-hydroxylase in mouse adrenal glands and in transfected Y1 adrenocortical tumor cells. Proc Natl Acad Sci USA 82:7860-7864
2. Handler JD, Schimmer BP, Flynn TR, Szyf M, Seidman JG, Parker KL 1988 An enhancer element and a functional cyclic AMP-de- pendent protein kinase are required for expression of adrenocor- tical 21-hydroxylase. J Biol Chem 263:13068-13073
3. Parker KL, Schimmer BP, Chaplin DD, Seidman JG 1986 Char- acterization of a regulatory region of the steroid 21-hydroxylase gene. J Biol Chem 261:15353-15355
4. Parker KL, Schimmer BP 1997 Steroidogenic factor 1: a key de- terminant of endocrine development and function. Endocr Rev 18:361-377
5. Lala DS, Rice DA, Parker KL 1992 Steroidogenic factor 1, a key regulator of steroidogenic enzyme expression, is the mouse ho- molog of Fushi Tarazu-Factor 1. Mol Endocrinol 6:1249-1258
6. Honda S, Morohashi K, Nomura M, Takeya H, Kitajima M, Omura T 1993 Ad4BP regulating steroidogenic P-450 gene is a member of steroid hormone recepter superfamily. J Biol Chem 268:7494-7502
7. Little TH, Zhang Y, Matulis CK, Weck J, Zhang Z, Ramachandran A, Mayo KE, Radhakrishnan I 2006 Sequence-specific deoxyribonu- cleic acid (DNA) recognition by steroidogenic factor 1: a helix at the carboxy terminus of the DNA binding domain is necessary for com- plex stability. Mol Endocrinol 20:831-843
8. Ikeda Y, Lala DS, Luo X, Kim E, Moisan MP, Parker KL 1993 Characterization of the mouse FTZ-F1 gene, which encodes a key regulator of steroid hydroxylase gene expression. Mol Endocrinol 7:852-860
9. Morohashi K, Iida H, Nomura M, Hatano O, Honda S, Tsukiyama T, Niwa O, Hara T, Takakusu A, Shibata Y, Omura T 1994 Functional difference between Ad4BP and ELP, and their distributions in steroidogenic tissues. Mol Endocrinol 8:643-653
10. Morohashi Ki 1999 Gonadal and extragonadal functions of Ad4BP/SF-1: developmental aspects. Trends Endocrinol Metab 10:169-173
11. Ramayya MS, Zhou J, Kino T, Segars JH, Bondy CA, Chrousos GP 1997 Steroidogenic factor 1 messenger ribonucleic acid expres- sion in steroidogenic and nonsteroidogenic human tissues: North- ern blot and in situ hybridization studies. J Clin Endocrinol Metab 82:1799-1806
12. Barnhart KM, Mellon PL 1994 The orphan nuclear receptor, ste- roidogenic factor-1, regulates the glycoprotein hormone @-subunit gene in pituitary gonadotropes. Mol Endocrinol 8:878-885
13. Ingraham HA, Lala DS, Ikeda Y, Luo X, Shen WH, Nachtigal MW, Abbud R, Nilson JH, Parker KL 1994 The nuclear receptor steroidogenic factor 1 acts at multiple levels of the reproductive axis. Genes Dev 8:2302-2312
14. Ngan ES, Cheng PK, Leung PC, Chow BK 1999 Steroidogenic factor-1 interacts with a gonadotrope-specific element within the first exon of the human gonadotropin-releasing hormone receptor gene to mediate gonadotrope-specific expression. Endocrinology 140:2452-2462
15. Morohashi K, Tsuboi-Asai H, Matsushita S, Suda M, Nakashima M, Sasano H, Hataba Y, Li CL, Fukata J, Irie J, Watanabe T, Nagura H, Li E 1999 Structural and functional abnormalities in the spleen of an mFTZ-F1 gene-disrupted mouse. Blood 93:1586- 1594
16. Wehrenberg U, Prange-Kiel J, Rune GM 2001 Steroidogenic factor-1 expression in marmoset and rat hippocampus: co-localization with StAR and aromatase. J Neurochem 76:1879-1886
17. Hanley NA, Ball SG, Clement-Jones M, Hagan DM, Strachan T, Lindsay S, Robson S, Ostrer H, Parker KL, Wilson DI 1999 Ex- pression of steroidogenic factor 1 and Wilms’ tumour 1 during early human gonadal development and sex determination. Mech Dev 87:175-180
18. Hanley NA, Rainey WE, Wilson DI, Ball SG, Parker KL 2001 Expression profiles of SF-1, DAX1, and CYP17 in the human fetal adrenal gland: potential interactions in gene regulation. Mol En- docrinol 15:57-68
19. Ikeda Y, Shen WH, Ingraham HA, Parker KL 1994 Developmen- tal expression of mouse steroidogenic factor-1, an essential regu- lator of the steroid hydroxylases. Mol Endocrinol 8:654-662
20. Morohashi K, Hatano O, Nomura M, Takayama K, Hara M, Yoshii H, Takakusu A, Omura T 1995 Function and distribution
of a steroidogenic cell-specific transcription factor, Ad4BP. J Ste- roid Biochem Mol Biol 53:81-88
21. Shen JH, Ingraham HA 2002 Regulation of the orphan nuclear receptor steroidogenic factor 1 by Sox proteins. Mol Endocrinol 16:529-540
22. Daggett MA, Rice DA, Heckert LL 2000 Expression of steroido- genic factor 1 in the testis requires an E box and CCAAT box in its promoter proximal region. Biol Reprod 62:670-679
23. Harris AN, Mellon PL 1998 The basic helix-loop-helix, leucine zipper transcription factor, USF (upstream stimulatory factor), is a key regulator of SF-1 (steroidogenic factor-1) gene expression in pituitary gonadotrope and steroidogenic cells. Mol Endocrinol 12:714-726
24. Nomura M, Bartsch S, Nawata H, Omura T, Morohashi K 1995 An E box element is required for the expression of the ad4bp gene, a mammalian homologue of ftz-f1 gene, which is essential for adrenal and gonadal development. J Biol Chem 270:7453-7461
25. Scherrer SP, Rice DA, Heckert LL 2002 Expression of steroido- genic factor 1 in the testis requires an interactive array of elements within its proximal promoter. Biol Reprod 67:1509-1521
26. Woodson KG, Crawford PA, Sadovsky Y, Milbrandt J 1997 Char- acterization of the promoter of SF-1, an orphan nuclear receptor required for adrenal and gonadal development. Mol Endocrinol 11:117-126
27. Tamura M, Kanno Y, Chuma S, Saito T, Nakatsuji N 2001 Pod- 1/Capsulin shows a sex- and stage-dependent expression pattern in the mouse gonad development and represses expression of Ad4BP/SF-1. Mech Dev 102:135-144
28. Tremblay JJ, Viger RS 2001 GATA factors differentially activate multiple gonadal promoters through conserved GATA regulatory elements. Endocrinology 142:977-986
29. Wilhelm D, Englert C 2002 The Wilms tumor suppressor WT1 regulates early gonad development by activation of Sf1. Genes Dev 16:1839-1851
30. Sakai N, Terami H, Suzuki S, Haga M, Nomoto K, Tsuchida N, Morohashi K, Saito N, Asada M, Hashimoto M, Harada D, Asahara H, Ishikawa T, Shimada F, Sakurada K 2008 Identification of NR5A1 (SF-1/AD4BP) gene expression modulators by large-scale gain and loss of function studies. J Endocrinol 198:489-497
31. Katoh-Fukui Y, Owaki A, Toyama Y, Kusaka M, Shinohara Y, Maekawa M, Toshimori K, Morohashi K 2005 Mouse Polycomb M33 is required for splenic vascular and adrenal gland formation through regulating Ad4BP/SF1 expression. Blood 106:1612-1620
32. Biason-Lauber A, Konrad D, Meyer M, DeBeaufort C, Schoenle EJ 2009 Ovaries and female phenotype in a girl with 46,XY karyotype and mutations in the CBX2 gene. Am J Hum Genet 84:658-663
33. Stallings NR, Hanley NA, Majdic G, Zhao L, Bakke M, Parker KL 2002 Development of a transgenic green fluorescent protein lineage marker for steroidogenic factor 1. Mol Endocrinol 16:2360-2370
34. Karpova T, Presley J, Manimaran RR, Scherrer SP, Tejada L, Peterson KR, Heckert LL 2005 A FTZ-F1-containing yeast artifi- cial chromosome recapitulates expression of steroidogenic factor 1 in vivo. Mol Endocrinol 19:2549-2563
35. Shima Y, Zubair M, Ishihara S, Shinohara Y, Oka S, Kimura S, Okamoto S, Minokoshi Y, Suita S, Morohashi K 2005 Ventrome- dial hypothalamic nucleus-specific enhancer of Ad4BP/SF-1 gene. Mol Endocrinol 19:2812-2823
36. Shima Y, Zubair M, Komatsu T, Oka S, Yokoyama C, Tachibana T, Hjalt TA, Drouin J, Morohashi K 2008 Pituitary homeobox 2 regulates adrenal4 binding protein/steroidogenic factor-1 gene transcription in the pituitary gonadotrope through interaction with the intronic enhancer. Mol Endocrinol 22:1633-1646
37. Zubair M, Ishihara S, Oka S, Okumura K, Morohashi K 2006 Two-step regulation of Ad4BP/SF-1 gene transcription during fe- tal adrenal development: initiation by a Hox-Pbx1-Prep1 complex and maintenance via autoregulation by Ad4BP/SF-1. Mol Cell Biol 26:4111-4121
38. Zubair M, Oka S, Parker KL, Morohashi K 2009 Transgenic ex-
pression of Ad4BP/SF-1 in fetal adrenal progenitor cells leads to ectopic adrenal formation. Mol Endocrinol 23:1657-1667
39. Zubair M, Parker KL, Morohashi K 2008 Developmental links between the fetal and adult zones of the adrenal cortex revealed by lineage tracing. Mol Cell Biol 28:7030-7040
40. Xue Q, Lin Z, Yin P, Milad MP, Cheng YH, Confino E, Reierstad S, Bulun SE 2007 Transcriptional activation of steroidogenic factor-1 by hypomethylation of the 5’ CpG island in endometriosis. J Clin Endocrinol Metab 92:3261-3267
41. Hoivik EA, Aumo L, Aesoy R, Lillefosse H, Lewis AE, Perrett RM, Stallings NR, Hanley NA, Bakke M 2008 Deoxyribonucleic acid methylation controls cell type-specific expression of steroidogenic factor 1. Endocrinology 149:5599-5609
42. Mohn F, Schübeler D 2009 Genetics and epigenetics: stability and plasticity during cellular differentiation. Trends Genet 25:129-136
43. Ishihara SL, Morohashi K 2005 A boundary for histone acetyla- tion allows distinct expression patterns of the Ad4BP/SF-1 and GCNF loci in adrenal cortex cells. Biochem Biophys Res Commun 329:554-562
44. Sewer MB, Dammer EB, Jagarlapudi S 2007 Transcriptional reg- ulation of adrenocortical steroidogenic gene expression. Drug Metab Rev 39:371-388
45. Cao G, Zhao L, Stangl H, Hasegawa T, Richardson JA, Parker KL, Hobbs HH 1999 Developmental and hormonal regulation of murine scavenger receptor, class B, type 1. Mol Endocrinol 13:1460-1473
46. Lopez D, Sandhoff TW, McLean MP 1999 Steroidogenic factor-1 mediates cyclic 3’,5’-adenosine monophosphate regulation of the high density lipoprotein receptor. Endocrinology 140:3034-3044
47. Naville D, Barjhoux L, Jaillard C, Lebrethon MC, Saez JM, Bègeot M 1994 Characterization of the transcription start site of the ACTH receptor gene: presence of an intronic sequence in the 5’- flanking region. Mol Cell Endocrinol 106:131-135
48. Winnay JN, Hammer GD 2006 Adrenocorticotropic-mediated signaling cascades coordinate a cyclic pattern of steroidogenic factor-1-dependent transcriptional activation. Mol Endocrinol 20:147-166
49. Ito M, Park Y, Weck J, Mayo KE, Jameson JL 2000 Synergistic activation of the inhibin a-promoter by steroidogenic factor-1 and cyclic adenosine 3’,5’-monophosphate. Mol Endocrinol 14:66-81
50. Aesøy R, Mellgren G, Morohashi K, Lund J 2002 Activation of cAMP-dependent protein kinase increases the protein level of ste- roidogenic factor-1. Endocrinology 143:295-303
51. Lopez D, Nackley AC, Shea-Eaton W, Xue J, Schimmer BP, McLean MP 2001 Effects of mutating different steroidogenic factor-1 protein regions on gene regulation. Endocrine 14:353-362
52. Li D, Urs AN, Allegood J, Leon A, Merrill Jr AH, Sewer MB 2007 Cyclic AMP-stimulated interaction between steroidogenic factor 1 and diacylglycerol kinase 0 facilitates induction of CYP17. Mol Cell Biol 27:6669-6685
53. Urs AN, Dammer E, Sewer MB 2006 Sphingosine regulates the transcription of CYP17 by binding to steroidogenic factor-1. En- docrinology 147:5249-5258
54. Chen WY, Juan LJ, Chung BC 2005 SF-1 (nuclear receptor 5A1) activity is activated by cyclic AMP via p300-mediated recruitment to active foci, acetylation, and increased DNA binding. Mol Cell Biol 25:10442-10453
55. Fan W, Yanase T, Wu Y, Kawate H, Saitoh M, Oba K, Nomura M, Okabe T, Goto K, Yanagisawa J, Kato S, Takayanagi R, Nawata H 2004 Protein kinase A potentiates adrenal 4 binding protein/steroidogenic factor 1 transactivation by reintegrating the subcellular dynamic interactions of the nuclear receptor with its cofactors, general control nonderepressed-5/transformation/tran- scription domain-associated protein, and suppressor, dosage-sen- sitive sex reversal-1: a laser confocal imaging study in living KGN cells. Mol Endocrinol 18:127-141
56. Chau YM, Crawford PA, Woodson KG, Polish JA, Olson LM, Sadovsky Y 1997 Role of steroidogenic-factor 1 in basal and 3’,5’-
cyclic adenosine monophosphate-mediated regulation of cyto- chrome P450 side-chain cleavage enzyme in the mouse. Biol Re- prod 57:765-771
57. Nomura M, Kawabe K, Matsushita S, Oka S, Hatano O, Harada N, Nawata H, Morohashi K 1998 Adrenocortical and gonadal expression of the mammalian Ftz-F1 gene encoding Ad4BP/SF-1 is independent of pituitary control. J Biochem 124:217-224
58. Mamluk R, Greber Y, Meidan R 1999 Hormonal regulation of messenger ribonucleic acid expression for steroidogenic factor-1, steroidogenic acute regulatory protein, and cytochrome P450 side- chain cleavage in bovine luteal cells. Biol Reprod 60:628-634
59. Zhang P, Mellon SH 1996 The orphan nuclear receptor steroidogenic factor-1 regulates the cyclic adenosine 3’,5’-monophosphate-medi- ated transcriptional activation of rat cytochrome P450c17 (17a-hy- droxylase/c17-20 lyase). Mol Endocrinol 10:147-158
60. Hammer GD, Krylova I, Zhang Y, Darimont BD, Simpson K, Weigel NL, Ingraham HA 1999 Phosphorylation of the nuclear receptor SF-1 modulates cofactor recruitment: integration of hor- mone signaling in reproduction and stress. Mol Cell 3:521-526
61. Desclozeaux M, Krylova IN, Horn F, Fletterick RJ, Ingraham HA 2002 Phosphorylation and intramolecular stabilization of the li- gand binding domain in the nuclear receptor steroidogenic factor 1. Mol Cell Biol 22:7193-7203
62. Fowkes RC, Desclozeaux M, Patel MV, Aylwin SJ, King P, Ingraham HA, Burrin JM 2003 Steroidogenic factor-1 and the gonadotrope- specific element enhance basal and pituitary adenylate cyclase-acti- vating polypeptide-stimulated transcription of the human glycopro- tein hormone «-subunit gene in gonadotropes. Mol Endocrinol 17: 2177-2188
63. Lin BC, Suzawa M, Blind RD, Tobias SC, Bulun SE, Scanlan TS, Ingraham HA 2009 Stimulating the GPR30 estrogen receptor with a novel tamoxifen analogue activates SF-1 and promotes endome- trial cell proliferation. Cancer Res 69:5415-5423
64. Lewis AE, Rusten M, Hoivik EA, Vikse EL, Hansson ML, Wallberg AE, Bakke M 2008 Phosphorylation of steroidogenic factor 1 is me- diated by cyclin-dependent kinase 7. Mol Endocrinol 22:91-104
65. Angers S, Moon RT 2009 Proximal events in Wnt signal trans- duction. Nat Rev Mol Cell Biol 10:468-477
66. Kim AC, Reuter AL, Zubair M, Else T, Serecky K, Bingham NC, Lavery GG, Parker KL, Hammer GD 2008 Targeted disruption of ß-catenin in Sf1-expressing cells impairs development and main- tenance of the adrenal cortex. Development 135:2593-2602
67. Cohen Jr MM 2003 The hedgehog signaling network. Am J Med Genet A 123A:5-28
68. Philipp M, Caron MG 2009 Hedgehog signaling: is Smo a G protein-coupled receptor? Curr Biol 19:R125-R127
69. Yao HH, Whoriskey W, Capel B 2002 Desert Hedgehog/Patched 1 signaling specifies fetal Leydig cell fate in testis organogenesis. Genes Dev 16:1433-1440
70. Barsoum IB, Bingham NC, Parker KL, Jorgensen JS, Yao HH 2009 Activation of the Hedgehog pathway in the mouse fetal ovary leads to ectopic appearance of fetal Leydig cells and female pseudohermaphroditism. Dev Biol 329:96-103
71. Wilson TE, Fahrner TJ, Milbrandt J 1993 The orphan receptors NGFI-B and steroidogenic factor 1 establish monomer binding as a third paradigm of nuclear receptor-DNA interaction. Mol Cell Biol 13:5794-5804
72. Hammer GD, Parker KL, Schimmer BP 2005 Transcriptional regulation of adrenocortical development. Endocrinology 146: 1018-1024
73. Dammer EB, Leon A, Sewer MB 2007 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 21:415-438
74. Parker KL, Schimmer BP 2002 Genes essential for early events in gonadal development. Ann Med 34:171-178
75. Ikeda Y, Swain A, Weber TJ, Hentges KE, Zanaria E, Lalli E,
Tamai KT, Sassone-Corsi P, Lovell-Badge R, Camerino G, Parker KL 1996 Steroidogenic factor 1 and Dax-1 colocalize in multiple cell lineages: potential links in endocrine development. Mol Endo- crinol 10:1261-1272
76. Ito M, Yu R, Jameson JL 1997 DAX-1 inhibits SF-1-mediated transactivation via a carboxy-terminal domain that is deleted in adrenal hypoplasia congenita. Mol Cell Biol 17:1476-1483
77. Suzuki T, Kasahara M, Yoshioka H, Morohashi K, Umesono K 2003 LXXLL-related motifs in Dax-1 have target specificity for the orphan nuclear receptors Ad4BP/SF-1 and LRH-1. Mol Cell Biol 23:238-249
78. Crawford PA, Dorn C, Sadovsky Y, Milbrandt J 1998 Nuclear receptor DAX-1 recruits nuclear receptor corepressor N-CoR to steroidogenic factor 1. Mol Cell Biol 18:2949-2956
79. Gummow BM, Scheys JO, Cancelli VR, Hammer GD 2006 Recip- rocal regulation of a glucocorticoid receptor-steroidogenic factor-1 transcription complex on the Dax-1 promoter by glucocorticoids and adrenocorticotropic hormone in the adrenal cortex. Mol Endocrinol 20:2711-2723
80. Ragazzon B, Lefrançcois-Martinez AM, Val P, Sahut-Barnola I, Tournaire C, Chambon C, Gachancard-Bouya JL, Begue RJ, Veyssière G, Martinez A 2006 Adrenocorticotropin-dependent changes in SF-1/DAX-1 ratio influence steroidogenic genes expres- sion in a novel model of glucocorticoid-producing adrenocortical cell lines derived from targeted tumorigenesis. Endocrinology 147:1805- 1818
81. Xu B, Yang WH, Gerin I, Hu CD, Hammer GD, Koenig RJ 2009 Dax-1 and steroid receptor RNA activator (SRA) function as tran- scriptional coactivators for steroidogenic factor 1 in steroidogen- esis. Mol Cell Biol 29:1719-1734
82. Komatsu T, Mizusaki H, Mukai T, Ogawa H, Baba D, Shirakawa M, Hatakeyama S, Nakayama KI, Yamamoto H, Kikuchi A, Morohashi K 2004 Small ubiquitin-like modifier 1 (SUMO-1) modification of the synergy control motif of Ad4 binding protein/ steroidogenic factor 1 (Ad4BP/SF-1) regulates synergistic tran- scription between Ad4BP/SF-1 and Sox9. Mol Endocrinol 18: 2451-2462
83. Chen WY, Lee WC, Hsu NC, Huang F, Chung BC 2004 SUMO modification of repression domains modulates function of nuclear receptor 5A1 (steroidogenic factor-1). J Biol Chem 279:38730- 38735
84. Lee MB, Lebedeva LA, Suzawa M, Wadekar SA, Desclozeaux M, Ingraham HA 2005 The DEAD-box protein DP103 (Ddx20 or Gemin-3) represses orphan nuclear receptor activity via SUMO modification. Mol Cell Biol 25:1879-1890
85. Jacob AL, Lund J, Martinez P, Hedin L 2001 Acetylation of steroi- dogenic factor 1 protein regulates its transcriptional activity and re- cruits the coactivator GCN5. J Biol Chem 276:37659-37664
86. Garcia-Dominguez M, Reyes JC 2009 SUMO association with repressor complexes, emerging routes for transcriptional control. Biochim Biophys Acta 1789:451-459
87. Campbell LA, Faivre EJ, Show MD, Ingraham JG, Flinders J, Gross JD, Ingraham HA 2008 Decreased recognition of SUMO- sensitive target genes following modification of SF-1 (NR5A1). Mol Cell Biol 28:7476-7486
88. Yang WH, Heaton JH, Brevig H, Mukherjee S, Iñiguez-Lluhí JA, Hammer GD 2009 SUMOylation inhibits SF-1 activity by reduc- ing CDK7-mediated serine 203 phosphorylation. Mol Cell Biol 29:613-625
89. Tremblay JJ, Robert NM, Laguë E 2009 Nuclear receptors, tes- tosterone, and posttranslational modifications in human INSL3 promoter activity in testicular Leydig cells. Ann NY Acad Sci 1160:205-212
90. Lala DS, Syka PM, Lazarchik SB, Mangelsdorf DJ, Parker KL, Heyman RA 1997 Activation of the orphan nuclear receptor ste- roidogenic factor 1 by oxysterols. Proc Natl Acad Sci USA 94: 4895-4900
91. Christenson LK, McAllister JM, Martin KO, Javitt NB, Osborne TF,
Strauss III JF 1998 Oxysterol regulation of steroidogenic acute regu- latory protein gene expression. J Biol Chem 273:30729-30735
92. Mellon SH, Bair SR 1998 25-Hydroxycholesterol is not a ligand for the orphan nuclear receptor steroidogenic factor-1 (SF-1). Endocri- nology 139:3026-3029
93. Krylova IN, Sablin EP, Moore J, Xu RX, Waitt GM, Mackay JA, Juzumiene D, Bynum JM, Madauss K, Montana V, Lebedeva L, Suzawa M, Williams JD, Williams SP, Guy RK, Thornton JW, Fletterick RJ, Willson TM, Ingraham HA 2005 Structural analyses reveal phosphatidyl inositols as ligands for the NR5 orphan recep- tors SF-1 and LRH-1. Cell 120:343-355
94. Li Y, Choi M, Cavey G, Daugherty J, Suino K, Kovach A, Bingham NC, Kliewer SA, Xu HE 2005 Crystallographic identification and functional characterization of phospholipids as ligands for the orphan nuclear receptor steroidogenic factor-1. Mol Cell 17:491-502
95. Sablin EP, Blind RD, Krylova IN, Ingraham JG, Cai F, Williams JD, Fletterick RJ, Ingraham HA 2009 Structure of SF-1 bound by different phospholipids: evidence for regulatory ligands. Mol En- docrinol 23:25-34
96. Wang W, Zhang C, Marimuthu A, Krupka HI, Tabrizizad M, Shelloe R, Mehra U, Eng K, Nguyen H, Settachatgul C, Powell B, Milburn MV, West BL 2005 The crystal structures of human steroidogenic factor-1 and liver receptor homologue-1. Proc Natl Acad Sci USA 102:7505-7510
97. Whitby RJ, Dixon S, Maloney PR, Delerive P, Goodwin BJ, Parks DJ, Willson TM 2006 Identification of small molecule agonists of the orphan nuclear receptors liver receptor homolog-1 and steroi- dogenic factor-1. J Med Chem 49:6652-6655
98. Del Tredici AL, Andersen CB, Currier EA, Ohrmund SR, Fairbain LC, Lund BW, Nash N, Olsson R, Piu F 2008 Identification of the first synthetic steroidogenic factor 1 inverse agonists: pharmaco- logical modulation of steroidogenic enzymes. Mol Pharmacol 73: 900-908
99. Aigueperse C, Val P, Pacot C, Darne C, Lalli E, Sassone-Corsi P, Veyssiere G, Jean C, Martinez A 2001 SF-1 (steroidogenic factor- 1), C/EBPB (CCAAT/enhancer binding protein), and ubiquitous transcription factors NF1 (nuclear factor 1) and Sp1 (selective promoter factor 1) are required for regulation of the mouse aldose reductase-like gene (AKR1B7) expression in adrenocortical cells. Mol Endocrinol 15:93-111
100. Rui X, Tsao J, Scheys JO, Hammer GD, Schimmer BP 2008 Con- tributions of specificity protein-1 and steroidogenic factor 1 to Adcy4 expression in Y1 mouse adrenal cells. Endocrinology 149: 3668-3678
101. Bassett MH, Zhang Y, Clyne C, White PC, Rainey WE 2002 Differential regulation of aldosterone synthase and 11ß-hydrox- ylase transcription by steroidogenic factor-1. J Mol Endocrinol 28:125-135
102. Frigeri C, Tsao J, Czerwinski W, Schimmer BP 2000 Impaired steroidogenic factor 1 (NR5A1) activity in mutant Y1 mouse ad- renocortical tumor cells. Mol Endocrinol 14:535-544
103. Tremblay JJ, Robert NM 2005 Role of nuclear receptors in INSL3 gene transcription in Leydig cells. Ann NY Acad Sci 1061:183-189
104. Zimmermann S, Schwärzler A, Buth S, Engel W, Adham IM 1998 Transcription of the Leydig insulin-like gene is mediated by ste- roidogenic factor-1. Mol Endocrinol 12:706-713
105. Fynn-Thompson E, Cheng H, Teixeira J 2003 Inhibition of ste- roidogenesis in Leydig cells by Mullerian-inhibiting substance. Mol Cell Endocrinol 211:99-104
106. Hinshelwood MM, Repa JJ, Shelton JM, Richardson JA, Mangelsdorf DJ, Mendelson CR 2003 Expression of LRH-1 and SF-1 in the mouse ovary: localization in different cell types correlates with differing function. Mol Cell Endocrinol 207:39-45
107. Peng N, Kim JW, Rainey WE, Carr BR, Attia GR 2003 The role of the orphan nuclear receptor, liver receptor homologue-1, in the regulation of human corpus luteum 3ß-hydroxysteroid dehydro- genase type II. J Clin Endocrinol Metab 88:6020-6028
108. Crawford PA, Sadovsky Y, Milbrandt J 1997 Nuclear receptor steroidogenic factor 1 directs embryonic stem cells toward the steroidogenic lineage. Mol Cell Biol 17:3997-4006
109. Gondo S, Yanase T, Okabe T, Tanaka T, Morinaga H, Nomura M, Goto K, Nawata H 2004 SF-1/Ad4BP transforms primary long-term cultured bone marrow cells into ACTH-responsive ste- roidogenic cells. Genes Cells 9:1239-1247
110. Tanaka T, Gondo S, Okabe T, Ohe K, Shirohzu H, Morinaga H, Nomura M, Tani K, Takayanagi R, Nawata H, Yanase T 2007 Steroidogenic factor 1/adrenal 4 binding protein transforms human bone marrow mesenchymal cells into steroidogenic cells. J Mol Endocrinol 39:343-350
111. Gondo S, Okabe T, Tanaka T, Morinaga H, Nomura M, Takayanagi R, Nawata H, Yanase T 2008 Adipose tissue-de- rived and bone marrow-derived mesenchymal cells develop into different lineage of steroidogenic cells by forced expression of steroidogenic factor 1. Endocrinology 149:4717-4725
112. Giuili G, Shen WH, Ingraham HA 1997 The nuclear receptor SF-1 mediates sexually dimorphic expression of Mullerian inhibiting substance, in vivo. Development 124:1799-1807
113. Arango NA, Lovell-Badge R, Behringer RR 1999 Targeted mu- tagenesis of the endogenous mouse Mis gene promoter: in vivo definition of genetic pathways of vertebrate sexual development. Cell 99:409-419
114. Keri RA, Nilson JH 1996 A steroidogenic factor-1 binding site is required for activity of the luteinizing hormone ß subunit promoter in gonadotropes of transgenic mice. J Biol Chem 271:10782-10785
115. Halvorson LM, Kaiser UB, Chin WW 1996 Stimulation of lutein- izing hormone ß gene promoter activity by the orphan nuclear receptor, steroidogenic factor-1. J Biol Chem 271:6645-6650
116. Duval DL, Nelson SE, Clay CM 1997 A binding site for steroido- genic factor-1 is part of a complex enhancer that mediates expres- sion of the murine gonadotropin-releasing hormone receptor gene. Biol Reprod 56:160-168
117. Jacobs SB, Coss D, McGillivray SM, Mellon PL 2003 Nuclear factor Y and steroidogenic factor 1 physically and functionally inter- act to contribute to cell-specific expression of the mouse follicle- stimulating hormone-ß gene. Mol Endocrinol 17:1470-1483
118. Kurrasch DM, Cheung CC, Lee FY, Tran PV, Hata K, Ingraham HA 2007 The neonatal ventromedial hypothalamus transcrip- tome reveals novel markers with spatially distinct patterning. J Neurosci 27:13624-13634
119. Luo X, Ikeda Y, Parker KL 1994 A cell-specific nuclear receptor is essential for adrenal and gonadal development and sexual differ- entiation. Cell 77:481-490
120. Sadovsky Y, Crawford PA, Woodson KG, Polish JA, Clements MA, Tourtellotte LM, Simburger K, Milbrandt J 1995 Mice defi- cient in the orphan receptor steroidogenic factor 1 lack adrenal glands and gonads but express P450 side-chain-cleavage enzyme in the placenta and have normal embryonic serum levels of corti- costeroids. Proc Natl Acad Sci USA 92:10939-10943
121. Bland ML, Jamieson CA, Akana SF, Bornstein SR, Eisenhofer G, Dallman MF, Ingraham HA 2000 Haploinsufficiency of steroido- genic factor-1 in mice disrupts adrenal development leading to an impaired stress response. Proc Natl Acad Sci USA 97:14488-14493
122. Shinoda K, Lei H, Yoshii H, Nomura M, Nagano M, Shiba H, Sasaki H, Osawa Y, Ninomiya Y, Niwa O, Morohashi KI, Li E 1995 Developmental defects of the ventromedial hypothalamic nucleus and pituitary gonadotroph in the Ftz-F1 disrupted mice. Dev Dyn 204:22-29
123. Ikeda Y, Luo X, Abbud R, Nilson JH, Parker KL 1995 The nuclear receptor steroidogenic factor 1 is essential for the formation of the ventromedial hypothalamic nucleus. Mol Endocrinol 9:478-486
124. Zhao L, Bakke M, Krimkevich Y, Cushman LJ, Parlow AF, Camper SA, Parker KL 2001 Steroidogenic factor 1 (SF1) is essential for pituitary gonadotrope function. Development 128:147-154
125. Jeyasuria P, Ikeda Y, Jamin SP, Zhao L, De Rooij DG, Themmen AP, Behringer RR, Parker KL 2004 Cell-specific knockout of ste-
roidogenic factor 1 reveals its essential roles in gonadal function. Mol Endocrinol 18:1610-1619
126. Pelusi C, Ikeda Y, Zubair M, Parker KL 2008 Impaired follicle development and infertility in female mice lacking steroidogenic factor 1 in ovarian granulosa cells. Biol Reprod 79:1074-1083
127. Zhao L, Kim KW, Ikeda Y, Anderson KK, Beck L, Chase S, Tobet SA, Parker KL 2008 Central nervous system-specific knockout of steroidogenic factor 1 results in increased anxiety-like behavior. Mol Endocrinol 22:1403-1415
128. Canteras NS 2002 The medial hypothalamic defensive system: hodological organization and functional implications. Pharmacol Biochem Behav 71:481-491
129. Tran PV, Akana SF, Malkovska I, Dallman MF, Parada LF, Ingraham HA 2006 Diminished hypothalamic bdnf expression and impaired VMH function are associated with reduced SF-1 gene dosage. J Comp Neurol 498:637-648
130. Kim KW, Zhao L, Parker KL 2009 Central nervous system-spe- cific knockout of steroidogenic factor 1. Mol Cell Endocrinol 300: 132-136
131. Achermann JC, Ito M, Ito M, Hindmarsh PC, Jameson JL 1999 A mutation in the gene encoding steroidogenic factor-1 causes XY sex reversal and adrenal failure in humans. Nat Genet 22:125-126
132. Biason-Lauber A, Schoenle EJ 2000 Apparently normal ovarian differentiation in a prepubertal girl with transcriptionally inactive steroidogenic factor 1 (NR5A1/SF-1) and adrenocortical insuffi- ciency. Am J Hum Genet 67:1563-1568
133. Achermann JC, Ozisik G, Ito M, Orun UA, Harmanci K, Gurakan B, Jameson JL 2002 Gonadal determination and adrenal develop- ment are regulated by the orphan nuclear receptor steroidogenic factor-1, in a dose-dependent manner. J Clin Endocrinol Metab 87:1829-1833
134. Köhler B, Lin L, Ferraz-de-Souza B, Wieacker P, Heidemann P, Schröder V, Biebermann H, Schnabel D, Grüters A, Achermann JC 2008 Five novel mutations in steroidogenic factor 1 (SF1, NR5A1) in 46,XY patients with severe underandrogenization but without adrenal insufficiency. Hum Mutat 29:59-64
135. Lourenço D, Brauner R, Lin L, De Perdigo A, Weryha G, Muresan M, Boudjenah R, Guerra-Junior G, Maciel-Guerra AT, Achermann JC, McElreavey K, Bashamboo A 2009 Mutations in NR5A1 asso- ciated with ovarian insufficiency. N Engl J Med 360:1200-1210
136. Bulun SE, Utsunomiya H, Lin Z, Yin P, Cheng YH, Pavone ME, Tokunaga H, Trukhacheva E, Attar E, Gurates B, Milad MP, Confino E, Su E, Reierstad S, Xue Q 2009 Steroidogenic factor-1 and endometriosis. Mol Cell Endocrinol 300:104-108
137. Figueiredo BC, Cavalli LR, Pianovski MA, Lalli E, Sandrini R, Ribeiro RC, Zambetti G, DeLacerda L, Rodrigues GA, Haddad BR 2005 Amplification of the steroidogenic factor 1 gene in child- hood adrenocortical tumors. J Clin Endocrinol Metab 90:615-619
138. Pianovski MA, Cavalli LR, Figueiredo BC, Santos SC, Doghman M, Ribeiro RC, Oliveira AG, Michalkiewicz E, Rodrigues GA, Zambetti G, Haddad BR, Lalli E 2006 SF-1 overexpression in childhood adrenocortical tumours. Eur J Cancer 42:1040-1043
139. Jorgensen JS, Nilson JH 2001 AR suppresses transcription of the LHß subunit by interacting with steroidogenic factor-1. Mol En- docrinol 15:1505-1516
140. Li LA, Chiang EF, Chen JC, Hsu NC, Chen YJ, Chung BC 1999 Function of steroidogenic factor 1 domains in nuclear localization, transactivation, and interaction with transcription factor TFIIB and c-Jun. Mol Endocrinol 13:1588-1598
141. Nachtigal MW, Hirokawa Y, Enyeart-VanHouten DL, Flanagan JN, Hammer GD, Ingraham HA 1998 Wilms’ tumor 1 and Dax-1 modulate the orphan nuclear receptor SF-1 in sex-specific gene expression. Cell 93:445-454
142. Tremblay JJ, Drouin J 1999 Egr-1 is a downstream effector of GnRH and synergizes by direct interaction with Ptx1 and SF-1 to enhance luteinizing hormone ß gene transcription. Mol Cell Biol 19:2567-2576
143. Wang DS, Kobayashi T, Zhou LY, Paul-Prasanth B, Ijiri S, Sakai
F, Okubo K, Morohashi K, Nagahama Y 2007 Foxl2 up-regulates aromatase gene transcription in a female-specific manner by bind- ing to the promoter as well as interacting with ad4 binding protein/ steroidogenic factor 1. Mol Endocrinol 21:712-725
144. Tremblay JJ, Viger RS 1999 Transcription factor GATA-4 en- hances Mullerian inhibiting substance gene transcription through a direct interaction with the nuclear receptor SF-1. Mol Endocri- nol 13:1388-1401
145. Hong CY, Park JH, Seo KH, Kim JM, Im SY, Lee JW, Choi HS, Lee K 2003 Expression of MIS in the testis is downregulated by tumor necrosis factor « through the negative regulation of SF-1 transactivation by NF-KB. Mol Cell Biol 23:6000-6012
146. Tremblay JJ, Marcil A, Gauthier Y, Drouin J 1999 Ptx1 regulates SF-1 activity by an interaction that mimics the role of the ligand- binding domain. EMBO J 18:3431-3441
147. Schepers G, Wilson M, Wilhelm D, Koopman P 2003 SOX8 is ex- pressed during testis differentiation in mice and synergizes with SF1 to activate the Amh promoter in vitro. J Biol Chem 278:28101- 28108
148. De Santa Barbara P, Bonneaud N, Boizet B, Desclozeaux M, Moniot B, Sudbeck P, Scherer G, Poulat F, Berta P 1998 Direct interaction of SRY-related protein SOX9 and steroidogenic factor 1 regulates transcription of the human anti-Müllerian hormone gene. Mol Cell Biol 18:6653-6665
149. Liu Z, Simpson ER 1997 Steroidogenic factor 1 (SF-1) and SP1 are required for regulation of bovine CYP11A gene expression in bo- vine luteal cells and adrenal Y1 cells. Mol Endocrinol 11:127-137
150. Liu Z, Simpson ER 1999 Molecular mechanism for cooperation between Sp1 and steroidogenic factor-1 (SF-1) to regulate bovine CYP11A gene expression. Mol Cell Endocrinol 153:183-196
151. Monté D, DeWitte F, Hum DW 1998 Regulation of the human P450scc gene by steroidogenic factor 1 is mediated by CBP/p300. J Biol Chem 273:4585-4591
152. Kabe Y, Goto M, Shima D, Imai T, Wada T, Morohashi K, Shirakawa M, Hirose S, Handa H 1999 The role of human MBF1 as a transcriptional coactivator. J Biol Chem 274:34196-34202
153. Børud B, Hoang T, Bakke M, Jacob AL, Lund J, Mellgren G 2002 The nuclear receptor coactivators p300/CBP/cointegrator-associ- ated protein (p/CIP) and transcription intermediary factor 2 (TIF2) differentially regulate PKA-stimulated transcriptional ac- tivity of steroidogenic factor 1. Mol Endocrinol 16:757-773
154. Zhou D, Quach KM, Yang C, Lee SY, Pohajdak B, Chen S 2000 PNRC: a proline-rich nuclear receptor coregulatory protein that modulates transcriptional activation of multiple nuclear receptors including orphan receptors SF1 (steroidogenic factor 1) and ERRa1 (estrogen related receptor @-1). Mol Endocrinol 14:986-998
155. Curtin D, Ferris HA, Häkli M, Gibson M, Jänne OA, Palvimo JJ, Shupnik MA 2004 Small nuclear RING finger protein stimulates the rat luteinizing hormone-beta promoter by interacting with Sp1 and steroidogenic factor-1 and protects from androgen suppres- sion. Mol Endocrinol 18:1263-1276
156. Crawford PA, Polish JA, Ganpule G, Sadovsky Y 1997 The activa- tion function-2 hexamer of steroidogenic factor-1 is required, but not sufficient for potentiation by SRC-1. Mol Endocrinol 11:1626-1635
157. Ito M, Yu RN, Jameson JL 1998 Steroidogenic factor-1 contains a carboxy-terminal transcriptional activation domain that interacts with steroid receptor coactivator-1. Mol Endocrinol 12:290-301
158. Gizard F, Lavallee B, De Witte F, Teissier E, Staels B, Hum DW 2002 The transcriptional regulating protein of 132 kDa (TReP-
132) enhances P450scc gene transcription through interaction with steroidogenic factor-1 in human adrenal cells. J Biol Chem 277:39144-39155
159. Dammer EB, Sewer MB 2008 Phosphorylation of CtBP1 by cAMP- dependent protein kinase modulates induction of CYP17 by stimu- lating partnering of CtBP1 and 2. J Biol Chem 283:6925-6934
160. Ou Q, Mouillet JF, Yan X, Dorn C, Crawford PA, Sadovsky Y 2001 The DEAD box protein DP103 is a regulator of steroido- genic factor-1. Mol Endocrinol 15:69-79
161. Park YY, Park KC, Shong M, Lee SJ, Lee YH, Choi HS 2007 EID-1 interacts with orphan nuclear receptor SF-1 and represses its transactivation. Mol Cells 24:372-377
162. Song KH, Park YY, Kee HJ, Hong CY, Lee YS, Ahn SW, Kim HJ, Lee K, Kook H, Lee IK, Choi HS 2006 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 281:15605-15614
163. Mellgren G, Børud B, Hoang T, Yri OE, Fladeby C, Lien EA, Lund J 2003 Characterization of receptor-interacting protein RIP140 in the regulation of SF-1 responsive target genes. Mol Cell Endocrinol 203:91-103
164. Sugawara T, Abe S, Sakuragi N, Fujimoto Y, Nomura E, Fujieda K, Saito M, Fujimoto S 2001 RIP 140 modulates transcription of the steroidogenic acute regulatory protein gene through interac- tions with both SF-1 and DAX-1. Endocrinology 142:3570-3577
165. Borud B, Mellgren G, Lund J, Bakke M 2003 Cloning and characterization of a novel zinc finger protein that modulates the transcriptional activity of nuclear receptors. Mol Endocri- nol 17:2303-2319
166. Hossain A, Saunders GF 2003 Synergistic cooperation between the B-catenin signaling pathway and steroidogenic factor 1 in the activation of the Mullerian inhibiting substance type II receptor. J Biol Chem 278:26511-26516
167. Gummow BM, Winnay JN, Hammer GD 2003 Convergence of Wnt signaling and steroidogenic factor-1 (SF-1) on transcription of the rat inhibin & gene. J Biol Chem 278:26572-26579
168. Mizusaki H, Kawabe K, Mukai T, Ariyoshi E, Kasahara M, Yoshioka H, Swain A, Morohashi K 2003 Dax-1 (dosage-sensitive sex reversal-adrenal hypoplasia congenita critical region on the X chromosome, gene 1) gene transcription is regulated by wnt4 in the female developing gonad. Mol Endocrinol 17:507-519
169. Parakh TN, Hernandez JA, Grammer JC, Weck J, Hunzicker- Dunn M, Zeleznik AJ, Nilson JH 2006 Follicle-stimulating hor- mone/cAMP regulation of aromatase gene expression requires B-catenin. Proc Natl Acad Sci USA 103:12435-12440
170. Jordan BK, Shen JH, Olaso R, Ingraham HA, Vilain E 2003 Wnt4 overexpression disrupts normal testicular vasculature and inhibits testosterone synthesis by repressing steroidogenic factor 1/B-cate- nin synergy. Proc Natl Acad Sci USA 100:10866-10871
171. Sewer MB, Nguyen VQ, Huang CJ, Tucker PW, Kagawa N, Waterman MR 2002 Transcriptional activation of human CYP17 in H295R adrenocortical cells depends on complex formation among p54(nrb)/NonO, protein-associated splicing factor, and SF-1, a complex that also participates in repression of transcrip- tion. Endocrinology 143:1280-1290
172. Mukai T, Kusaka M, Kawabe K, Goto K, Nawata H, Fujieda K, Morohashi K 2002 Sexually dimorphic expression of Dax-1 in the adrenal cortex. Genes Cells 7:717-729