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Department of Veterinary Physiology and Pharmacology, Texas A & M University, College Station, Texas 77843-4466
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ABSTRACT |
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 TOP ABSTRACT INTRODUCTIONRESULTSDISCUSSIONMATERIALs AND METHODSREFERENCES |
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INTRODUCTION |
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TOPABSTRACTINTRODUCTIONRESULTSDISCUSSIONMATERIALs AND METHODSREFERENCES |
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The molecular mechanism by which nuclear receptors modulate gene
transcription has, in its simplest form, been recognized for some time
(12). For example, 17ß-estradiol (E2) modulates
transcription by binding with the estrogen receptor (ER); the liganded
ER homodimerizes and binds to its cognate sequence, the estrogen
responsive element (ERE), in the promoter of a target gene, resulting
in either transcriptional activation or repression in a cellular and
promoter-specific manner (13, 14, 15). This selectivity by steroid hormone
receptors can be the result of ligand modification, such as differences
in pharmacokinetics and metabolism (16), or changes in the receptor
through splice variants (17, 18), or covalent modifications (2, 19, 20, 21, 22, 23). In the context of complex promoters, EREs are generally found
in multiple copies or encased among binding motifs for other
transcription factors. For example, analysis of 5'-promoter regions in
the c-myc and rat creatine kinase B (CKB) genes
identified estrogen-responsive regions that did not contain classic
palindromic EREs (1, 24). Dubik and Shiu (1) pointed out that both of
these promoter sequences contained Sp1 and ERE-half sites, and it was
suggested that transactivation of the c-myc and
CKB genes may be due to interactions between ER and Sp1
complexes, which are stabilized by interactions with an
Sp1(N)xERE half-site DNA-binding motif. Research in this
laboratory has identified a functional Sp1(N)23ERE
half-site in the noncoding strand of the cathepsin D gene (25, 26).
Using the Sp1(N)23ERE sequence in both gel mobility shift
and functional transient transfection studies in MCF-7 human breast
cancer cells and HeLa cells, it was shown that an ER/Sp1 complex binds
to the Sp1(N)23ERE oligonucleotide, and E2
induces reporter gene activity using a cathepsin D Sp1/ERE-CAT
construct. Although the results clearly demonstrated that a DNA-bound
ER/Sp1 complex was formed, the nature of this interaction and the
involvement of other proteins was not determined. Subsequent
studies have also demonstrated that Sp1(N)xERE motifs also
play a role in estrogen-regulated retinoic acid receptor 
(RAR)
and heat shock protein 27 (Hsp 27) gene expression in human breast
cancer cells (27, 28). Thus, the cooperative interactions of Sp1 and ER
proteins play a role in regulation of at least five estrogen-inducible
genes, including c-myc, CKB, cathepsin D, RAR,
and Hsp 27, and requires the presence of Sp1 and ERE half-site motifs
in which there is considerable variability in the ERE half-site
sequences, their orientation, and the number of intervening nucleotides
between Sp1 and ERE DNA-binding sites. This study reports that estrogen
induces reporter gene activity in MCF-7 cells transiently transfected
with a human ER expression plasmid and constructs containing GC-rich
Sp1-binding sequences linked to a bacterial chloramphenicol acetyl
transferase (CAT) reporter gene. In gel mobility shift assays, ER
enhanced Sp1 binding to 32P-labeled oligonucleotides
containing GC-rich binding sites, and, in the absence of DNA,
[35S]ER and Sp1 proteins could be coimmunoprecipated with
Sp1 antibodies. Thus, the functional synergy between ER and Sp1 is
associated with protein-protein interactions and represents an
estrogen-induced transactivation pathway that does not require ER-DNA
binding.
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RESULTS |
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TOPABSTRACTINTRODUCTIONRESULTSDISCUSSIONMATERIALs AND METHODSREFERENCES |
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compare the
induction of CAT activity by E2 in MCF-7 cells transiently
transfected with wild type Hsp-CATs Sp1/ERE (lanes 1 and 2) and
Hsp-CAT
Sp1/ERE (lanes 7 and 8) plasmids and with the corresponding
mutant plasmids Hsp-CATs Sp1/ERE and Hsp-CAT
Sp1/ERE plasmids, which contain mutations in the ERE
half-sites. The results show that E2 induced CAT activity
using both wild type (lanes 1, 2, 7, and 8) and ERE mutant (lanes 3, 4,
9, and 10) plasmids, suggesting that hormone responsiveness of these
constructs did not require an intact ERE half-site. Moreover,
E2 induced CAT activity in cells transfected with the
Sp1-CAT(Hsp) plasmid, which contains only the Sp1 oligonucleotide
insert from the Hsp 27 gene promoter. The role of Sp1 DNA-binding sites
on E2-responsiveness was confirmed in MCF-7 cells
transiently cotransfected with a construct containing an consensus Sp1
element (Sp1-TATA-CAT) and human ER (hER) expression plasmids.
E2 caused a concentration-dependent induction of CAT
activity (1.4- to 10.6-fold) (Fig. 2, lanes 1 through 4), and the effects were similar to those observed for
wild-type Hsp-CATs Sp1/ERE and Hsp-CAT Sp1/ERE plasmids and
constructs containing mutations in their ERE half-sites (Fig. 1).
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, lanes 1 through 4). In the presence of
in vitro translated ER, the dose-dependent increase in
retarded band formation by the same concentration gradient of Sp1
protein was further enhanced (Fig. 3, lanes 5 through 8). The intensity
of the ER-enhanced retarded band (lane 8) was significantly decreased
by competition with a 100-fold excess of unlabeled Sp1 oligonucleotide
(lane 10), but not by a 100-fold excess of wild type or mutant ERE
oligonucleotides (lanes 9 and 10, respectively). Intensity of the
retarded band was not decreased in competition with 100-fold excess
wild type ERE (lane 9). The results summarized in Fig. 4 also demonstrate that Sp1 binding to
the mutated [32P]Hsp 27-Sp1/ERE short
oligonucleotide (lanes 1 through 4) is enhanced by the ER (lanes 5
through 8), and the retarded band intensity (lane 8) is decreased by
unlabeled Sp1 (lane 10) but not by mutant ERE oligonucleotides (lane
9). The pattern of enhanced Sp1-DNA binding by ER using GC-rich sites
in the Hsp 27 promoter was also observed using a consensus
[32P]Sp1 oligonucleotide (Fig. 5A). The intensity of the bound
Sp1-[32P]Sp1 band (lanes 2 through 4) was enhanced after
incubation with ER (lanes 6 through 8), decreased by competition with
100-fold excess unlabeled Sp1 oligonucleotide (lane 9), but not
affected by unlabeled mutant ERE (lane 10). The time-dependent
formation of the Sp1-DNA retarded band was determined in the presence
or absence of ER (Fig. 5B). The results show that binding was maximal
after 10 min, and the overall on rate was increased 2.0- to 3.2-fold
over the 15-min incubation period. In contrast, the time-dependent loss
of the Sp1-DNA complex was independent of ER, and the off rate for
decomposition of the complex was comparable in the presence or absence
of ER (data not shown). In all of the protein-DNA-binding studies
(Figs. 3 through 5) enhanced Sp1-DNA binding by ER was
hormone-independent, since comparable results were obtained in the
presence or absence of 10-8 M E2
(data not shown).
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compare the effects of wild type hER
and mutant 11C-ER, in which the ER-DNA-binding domain has been deleted,
on Sp1-[32P]Hsp 27-Sp1/ERE short retarded band formation.
Both wild type hER and 11C-ER (lanes 4 and 6, respectively) caused a
2-fold increase in formation of the retarded band, whereas the mutant
15C-ER and 19C-ER (containing C- and N- terminal deletions,
respectively) do not affect retarded band formation (lanes 8 and 10,
respectively). The 11C-ER interactions with Sp1 were observed in the
absence of E2, and addition of the hormone did not affect
the interaction (data not shown). These results indicate that the DNA-
binding domain of the ER is not required for enhancement of Sp1-DNA
complex formation. The binding data also complement transactivation
assays in ER-negative MDA-MBA-231 cells transiently transfected with
Hsp-CAT or Sp1-TATA-CAT (Fig. 7)
plasmids and hER or 11C-ER expression plasmids. CAT activity is not
induced by 10-8 M E2 using either
Hsp-CAT Sp1/ERE or Sp1-TATA-CAT plasmids, whereas hormone
inducibility is restored with both plasmids after cotransfection of
wild type or 11C-ER. In contrast, hormone inducibility was not observed
using 15C-ER or 19C-ER expression plasmids.
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demonstrate that ER can be reversibly
cross-linked to Sp1. The radiolabeled [35S]ER was
immunoprecipitated by both Sp1 and ER antibodies (Fig. 8, lanes 2 and
5, respectively); [35S]ER alone was immunoprecipitated
with ER antibodies (lane 4). Incubation of Sp1 plus
35S-labeled unprogrammed lysate (UPL) followed by
immunoprecipitation with ER or Sp1 antibodies (lanes 3 and 6,
respectively) did not give a radiolabeled protein. The Sp1 antibody
(Ab) did not immunoprecipitate [35S]ER alone (data not
shown). Experiments performed without the utilization of cross-linking
reagents did not co-precipitate Sp1, indicating that the
protein-protein interaction between Sp1 and ER is transient under the
conditions used (data not shown). These data show that the ER and Sp1
proteins physically interact and form a protein-protein complex in the
absence of DNA. Moreover, the interaction of ER and Sp1 proteins was
observed in the presence or absence of E2 and therefore was
hormone-independent.
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, lanes 3 and 5, respectively). As an
additional control, [35S]ER was incubated with GST alone,
and negligible binding was observed (lane 4). Sp1 and ER were shown to
interact through a GST-Sp1 fusion protein incubated with
[35S]ER (lane 6). To further examine interactions between
Sp1 and ER, truncated GST-Sp1 fusion proteins containing various
regions of the Sp1 were incubated with [35S]ER and
analyzed. Incubation of GST-Sp1 [amino acids (aa) 1–293] with
[35S]ER showed undetectable binding to
[35S]ER; however, incubation of GST-Sp1 (aa 1–621) and
GST-Sp1 (aa 622–788) with [35S]ER showed that both
fusion proteins bound to [35S]ER (lanes 8 and 9,
respectively). These results indicate an interaction of
[35S]ER with two different GST-Sp1 fusion proteins;
however, the major site of interaction of ER with Sp1 protein is
associated with the C-terminal DNA-binding domain of Sp1.
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DISCUSSION |
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TOPABSTRACTINTRODUCTIONRESULTSDISCUSSIONMATERIALs AND METHODSREFERENCES |
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, and Hsp 27 genes are also
sequences that bind ER in association with the Sp1 protein (1, 24, 25, 26, 27, 28).
For example, the results illustrated in Fig. 1
confirm the estrogen
inducibility of wild type Hsp-CATs Sp1/ERE and Hsp-CAT Sp1/ERE
plasmids (lanes 1, 2, 7, and 8), which contain the
Sp1(N)10ERE half-site (-108 to -84) identified in the Hsp
27 gene promoter (28). However, estrogen responsiveness was also
observed with the same plasmids containing mutations in the ERE
half-sites (lanes 3, 4, 9, and 10). These results suggested that
E2-induced transactivation may require only GC-rich
Sp1-binding sites, and the results illustrated in Figs. 2 through 5
demonstrate 1) E2-responsiveness of constructs or
oligonucleotides containing consensus Sp1 promoter sequences (Fig. 2),
and 2) enhanced Sp1-[32P]Sp1 oligonucleotide retarded
band formation after coincubation with ER protein (Figs. 3 through 5).
Interestingly, after incubation of Sp1 and ER proteins with
[32P]Sp1, a ternary Sp1/ER-[32P]Sp1 complex
is not observed in gel mobility shift assays; however, ER enhances the
on rate of Sp1-[32P]Sp1 formation (Fig. 5B). This type of
protein (ER) enhancement of protein-DNA complex formation is comparable
to results of other studies showing that HTLV-1 Tax, SREPB, and cyclin
D1 enhance bZIP, Sp1, and ER binding to their respective enhancer
sequences without forming a ternary complex (44, 45, 46). The interactions
of ER with Sp1 are also similar to the functional synergy previously
reported for Sp1 and the erythroid cell transcription factor, GATA, in
which GATA synergistically enhances Sp1-induced transactivation in
Schneider cells using Sp1-dependent promoters (47, 48, 49). Moreover, in
the same cell line, RB protein and E2F1 enhanced Sp1-induced responses
utilizing several different constructs containing functional Sp1-
binding sites (50, 51).
The functional synergy between the ER and Sp1 nuclear transcription
factors was further investigated using mutant ERs containing deletions
in the DNA-binding (11C-ER), N-terminal (19C-ER), and C-terminal
(15C-ER) domains. The latter two ER mutants did not enhance Sp1 binding
in gel mobility shift assays using wild type [32P]Hsp
27-Sp1/ERE, which contains the Sp1(N)10ERE half-site motif
from the Hsp 27 gene promoter (Fig. 6), or transactivation using
Hsp-CAT Sp1/ERE or cSp1-TATA-CAT plasmids in MDA-MB-231 breast
cancer cells (Fig. 7). In contrast, both wild type and 11C-ER enhance
both Sp1 binding and transactivation (Figs. 6 and 7), indicating that
deletion of the DNA-binding domain of ER (11C-ER) does not abolish the
synergy of the ER with Sp1. These results are similar to those reported
for ER-AP1 interactions in which the AP1 complex serves as a tether,
when bound to its cognate DNA element, to target steroid receptors such
as the ER in the absence of a consensus ERE (52). This functional
interaction also occurs independently of ER binding to DNA and
parallels results observed for Sp1-ER interactions (Figs. 6 and 7).
Both ER and Sp1 physically interact with other nuclear proteins
(45, 46, 47, 48, 49, 50, 51, 52), and the results illustrated in Fig. 8 show that an ER/Sp1
protein complex can be coimmunoprecipitated. The interaction of Sp1 and
ER proteins and enhancement of Sp1-DNA binding by ER (Figs. 3 through 5) was observed in the presence or absence of E2, whereas
transactivation was hormone-dependent (Figs. 1, 2, and 7). The effects
of E2 on interactions of ER with other nuclear proteins are
variable; for example, GATA-1 binds ER only in the presence of
E2 (47), whereas binding of cyclin D1 to the ER is
E2-independent (46). Cyclin D1 enhancement of ERE-dependent
reporter gene activity is also observed in the absence of hormone,
whereas ER-mediated inhibition of GATA-induced responses are
E2-dependent. The rationale for the hormone-independent or
dependent responses associated with ER interactions with other nuclear
proteins is unclear and requires further research. In this study,
[35S]ER also bound to GST-Sp1 fusion protein prebound to
GST-Sepharose beads. [35S]ER interacted with truncated
GST-Sp1 fusion proteins containing the C-terminal (aa 622 to 788) but
not the N-terminal region of the protein (Fig. 9). Similar results have
been reported previously for interactions of Sp1 with E2F1 and GATA-1
in which binding is also mediated through the C-terminal DNA-binding
domain of Sp1 (47, 51). Our results also indicated that
[35S]ER interacted with regions of the B (partial) and C
domains (aa 294 to 621) of Sp1 (Fig. 9), suggesting that interactions
of Sp1 with the ER may involve more than one domain of Sp1. Our results
showed that ER primarily interacted with the C-terminal region of Sp1,
and the importance of the weaker interactions with other domains in the
Sp1 protein requires further study.
The Sp1 protein plays a major role in regulating expression of diverse cellular and viral genes, most of which are not affected by hormones. Nevertheless, results of this study demonstrate that GC-rich binding sites are potential targets for ER-mediated transactivation. This suggests that hormone responsiveness via Sp1/ER interactions will be highly promoter- and cell-specific, and current studies in this laboratory are focused on identification of estrogen-responsive Sp1 enhancer sequences and their cell-specific hormone-induced transactivation.
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MATERIALs AND METHODS |
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TOPABSTRACTINTRODUCTIONRESULTSDISCUSSIONMATERIALs AND METHODSREFERENCES |
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The oligonucleotides structures and their descriptors are given below and used throughout the manuscript to identify the specific oligonucleotide. The Sp1 and ERE half-sites are underlined, and mutated bases are indicated with an asterisk. Hsp 27-Sp1/ERE Short Oligo (Sense Strand):
5'-AGCTTGGAGGGGCGGCCCTCAAACGGGTCATTGCG-3' Hsp 27-Sp1/ERE short oligo (sense strand):
5'-AGCTTGGAGGGGCGGCCCTCAAACGA*A*TCATTGCG-3' Hsp 27-Sp1 oligo (sense strand):
5'-AGCTTGGAGGGGCGGCCCTCG-3' consensus Sp1 oligo (sense strand):
5'-AGCTTATTCGATCGGGGCGGGGCGAGCG-3' Hsp 27-Sp1/ERE long oligo (sense strand):
5'-AGCTTGGAGGGGCGGCCCTCAAACGGGTCATTGC-CATTA
ATAGAGACCTCAAACACCGCCTGCTAAAAATACCCGA-CTGG
AGGAGCATAAAAGCGCAGCCGAGCCCAGCGCCCCGC-ACTT
TTCTGAGGT-3' Hsp 27-Sp1/ERE long oligo (sense strand):
5'-AGCTTGGAGGGGCGGCCCTCAAACGA*A*TCATTGC-CATTA
ATAGAGACCTCAAACACCGCCTGCTAAAAATACCCGA-CTGG
AGGAGCATAAAAGCGCAGCCGAGCCCAGCGCCCCG-CACTT
TTCTGAGGT-3' E1B-TATA oligo (sense strand):
5'-GATCCGTCGACGCTGTAGGGGTATATAATGGTTGC-GGATC-3'
Cloning
The pBLTATA-CAT plasmid was made by digesting the
pBLCAT2 vector with BamHI and XhoI to remove the
thymidine kinase promoter; the double-stranded E1B-TATA oligonucleotide
containing complementary 5'-overhangs was then inserted into the
corresponding sites. The wild type Hsp 27-Sp1/ERE short and mutant Hsp
27-Sp1/ERE short and consensus Sp1 oligonucleotides were
cloned into the pBLTATA-CAT at the HindIII and
BamHI sites as previously described (28) to give the
Hsp-CATs Sp1/ERE, Hsp-CATs Sp1/ERE, and Sp1-TATA-CAT
plasmids, respectively. Wild type Hsp-CAT Sp1/ERE and mutant
Hsp-Sp1/ERE-CAT plasmids were constructed using the Hsp
27-Sp1/ERE long and Hsp 27-Sp1/ERE long oligonucleotides
(see above) which were cloned into the
HindIII/BamHI site of pBLCAT2 as previously
reported (28). The thymidine kinase (TK) promoter was then removed by
digesting wild type Hsp-CAT Sp1/ERE and mutant Hsp-CAT
Sp1/ERE plasmids with BamHI and XbaI
and religating the complementary sites. Ligation products were
transformed into DH5 cells, and clones were verified by
sequencing.
Transient Transfection Assay
Cultured MCF-7 and MDA-MB-231 cells were transfected by the
calcium phosphate method with 10 µg reporter plasmid and 5 µg of
either the appropriate hER plasmid or empty construct (pCDNA3-Neo, In
Vitrogen, Inc., Carlsbad, CA) as a control. After 18 h, the media
was changed and the cells were treated with Me2SO (0.2%
total volume) or E2 (10-8 M) in
Me2SO for 44 h. Cells were then washed with PBS and
scraped from the plates. Cell lysates were prepared in 0.16 ml of 0.25
M Tris-HCl, pH 7.5, by three freeze-thaw-sonication cycles
(3 min each). Cell lysates were incubated at 56 C for 7 min to remove
endogenous deacetylase activity. CAT activity was determined using 0.2
mCi
d-threo-[dichloroacetyl-1-14C]chloramphenicol
and 4 mM acetyl-CoA as substrates. The protein
concentrations were determined using BSA as a standard. After TLC,
acetylated products were visualized and quantitated using a Betascope
603 Blot analyzer (Intelligenetics, Mountain View, CA). CAT activity
was calculated as the percentage of that observed in cells treated with
Me2SO alone (arbitrarily set at 100%), and results are
expressed as means ± SD. The experiments were carried
out at least three times for each treatment group.
Electrophoretic Mobility Shift Assays Using in Vitro
Translated Proteins
Plasmids containing the WT-ER, 11C-ER, 15C-ER, and 19C-ER were
used to in vitro transcribe and translate the corresponding
proteins in a rabbit reticulocyte lysate kit (Promega, Madison, WI).
Parallel reactions with [35S]methionine were also
performed to monitor translational efficiency and control for loading.
Gel electromobility shift assays were performed by assembling the
appropriate in vitro translated proteins in 1x binding
buffer (20 mM HEPES, 5% glycerol, 100 mM
potassium chloride, 5 mM magnesium chloride, 0.5
mM dithiothreitol, 1 mM EDTA in a final volume
of 25 µl). E2 was added to the reaction at a final
concentration of 20 nM and then incubated on ice for 15
min. Sp1 and the labeled oligonucleotides (30,000 cpm) were then added
to the reaction mixtures in the presence of 1 µg
poly(deoxyinosinic-deoxycytidylic)acid, and the mixtures were incubated
for 15 min at 25 C. Samples were loaded onto a 4% polyacrylamide gel
(acrylamide-bisacrylamide ratio, 30:0.8) and run at 110 V in 0.09
M Tris-0.09 M borate-2 mM EDTA, pH
8.3. Protein-DNA binding was visualized by autoradiography and
quantitated by densitometry using the Scanalytics Zero-D software
package (Scanalytics, Billerica, MA) and a Sharp JX-330 scanner
(Mahwah, NJ).
Immunoprecipitation and Protein Cross-Linking
35S-labeled ER was synthesized and incubated with
Sp1 as described above. The reaction mixture was diluted with 1x
binding buffer and cross-linked with 5 mM
dithiobis(succinimidyl propionate), a bifunctional, reversible
cross-linker, for 1 h at 25 C, then quenched with 0.22
M lysine as described by Lin et al. (50).
Radioimmunoprecipitation (RIPA) was carried out by adding 500 µl of
RIPA buffer (PBS-1% NP40–0.5% sodium deoxycholate-0.1%DNA-10 mg/ml
phenylmethylsulfonyl fluoride (PMSF)-aprotinin 30 µl/ml-sodium
orthovanadate 10 µl/ml) and 1 µg of antisera. After incubating for
1 h at 4 C, 20 µl of Agarose A (Santa Cruz Biotechnology, Santa
Cruz) was added and incubated (rocking) for 1 h at 4 C. The bound
complex was then washed four times with RIPA buffer containing 2
M urea. The precipitated proteins were then eluted with 2X
SDS sample buffer to reverse the crosslinks and resolved on a 6%
SDS-polyacrylamide gel, dried and visualized by autoradiography.
GST Pulldown Experiment
GST, GST-Sp1, or GST-Sp1 (truncated) fusion proteins were
purified essentially as described by the manufacturer in GST: Gene
Fusion System (Pharmacia Biotech). DH5 bacterial cells transformed
with either pGEX-4T-1 or with plasmids pGEX-2TK-MCS-Sp1,
pGEX-2TK-MCS-Sp1 (1–293), pGEX-2TK-MCS-Sp1 (1 -621), or
pGEX-2TK-MCS-Sp1 (622–788) kindly provided by Professor Erhard
Wintersberger (51) were grown overnight in Lauria broth + 50 µg/ml
ampicillin at 37 C. Cultures were then diluted 1:10 with Lauria broth +
50 µg/ml ampicillin and grown at 37 C until A600 reached
between 0.5 to 0.7 (about 2–3 h). Protein expression was induced with
0.05 mM isopropyl-D-thiogalactopyranoside, and
cultures were allowed to grow for an additional 1.5 h; 1.5 ml were
transferred to an eppitube and centrifuged, and the pellet was
resuspended in 300 µl of sonication buffer [150 mM KCl,
40 mM HEPES (pH 7.5), 0.5 mM EDTA, 5.0
mM MgCl2, 1.0 mM dithiothreitol,
0.05% Nonidet P-40] supplemented with 1.0 mM PMSF, and 10
µg/µl aprotinin. Cells were lysed by sonication, and the crude
bacterial extract was either frozen at -80 C or used immediately. Ten
microliters of a 50% slurry of glutathione-Sepharose 4B beads were
added to bacterial extracts containing either GST, wild type, or
truncated GST-Sp1 fusion proteins and incubated at room
temperature for 30 min with shaking; the beads were then washed twice
with sonication buffer and once with hER wash buffer (250
mM NaCl, 0.1% Nonidet P-40, 50 mM HEPES (pH
7.5), 5.0 mM EDTA]. After the final wash, 80 µl of hER
binding buffer (hER wash buffer supplemented with 0.5 mM
dithiothreitol, 1.0 mM PMSF, 10 µg/µl aprotinin) and 3
µl of transcription and translation rabbit reticulocyte lysate system
(Promega) and in vitro translated hER were added to the
beads. This reaction was incubated at 4 C for 2 h and the beads
were washed four times with hER wash buffer. Ten microliters of SDS
loading buffer were added to the beads and heated at 100 C for 3 min,
and samples were analyzed on a 10% SDS polyacrylamide gel. Proteins
were fixed to the gel with 30% methanol/10% acetic acid solution for
20 min. The gel was then treated with EN3HANCE (DuPont),
dried and exposed to film.
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FOOTNOTES |
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This work was supported by NIH Grant ES-04176, the Robert A. Welch Foundation, and the Texas Agricultural Experiment Station.
Received for publication January 6, 1997. Revision received June 24, 1997. Accepted for publication July 17, 1997.
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TOPABSTRACTINTRODUCTIONRESULTSDISCUSSIONMATERIALs AND METHODSREFERENCES |
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 P. Hu, H. K. Kinyamu, L. Wang, J. Martin, T. K. Archer, and C. Teng Estrogen Induces Estrogen-related Receptor {alpha} Gene Expression and Chromatin Structural Changes in Estrogen Receptor (ER)-positive and ER-negative Breast Cancer Cells J. Biol. Chem., March 14, 2008; 283(11): 6752 - 6763. [Abstract] [Full Text] [PDF]
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K. J. Higgins, S. Liu, M. Abdelrahim, K. Vanderlaag, X. Liu, W. Porter, R. Metz, and S. Safe Vascular Endothelial Growth Factor Receptor-2 Expression Is Down-Regulated by 17{beta}-Estradiol in MCF-7 Breast Cancer Cells by Estrogen Receptor {alpha}/Sp Proteins Mol. Endocrinol., February 1, 2008; 22(2): 388 - 402. [Abstract] [Full Text] [PDF]
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P. Bagamasbad, K. L. Howdeshell, L. M. Sachs, B. A. Demeneix, and R. J. Denver A Role for Basic Transcription Element-binding Protein 1 (BTEB1) in the Autoinduction of Thyroid Hormone Receptor J. Biol. Chem., January 25, 2008; 283(4): 2275 - 2285. [Abstract] [Full Text] [PDF]
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C. D. DuSell, M. Umetani, P. W. Shaul, D. J. Mangelsdorf, and D. P. McDonnell 27-Hydroxycholesterol Is an Endogenous Selective Estrogen Receptor Modulator Mol. Endocrinol., January 1, 2008; 22(1): 65 - 77. [Abstract] [Full Text] [PDF]
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C. R. Cederroth, O. Schaad, P. Descombes, P. Chambon, J.-D. Vassalli, and S. Nef Estrogen Receptor {alpha} Is a Major Contributor to Estrogen-Mediated Fetal Testis Dysgenesis and Cryptorchidism Endocrinology, November 1, 2007; 148(11): 5507 - 5519. [Abstract] [Full Text] [PDF]
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S. Khan, F. Wu, S. Liu, Q. Wu, and S. Safe Role of specificity protein transcription factors in estrogen-induced gene expression in MCF-7 breast cancer cells J. Mol. Endocrinol., October 1, 2007; 39(4): 289 - 304. [Abstract] [Full Text] [PDF]
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D. Li, D. Mitchell, J. Luo, Z. Yi, S.-G. Cho, J. Guo, X. Li, G. Ning, X. Wu, and M. Liu Estrogen Regulates KiSS1 Gene Expression through Estrogen Receptor {alpha} and SP Protein Complexes Endocrinology, October 1, 2007; 148(10): 4821 - 4828. [Abstract] [Full Text] [PDF]
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 R. Martin, M. B. Taylor, G. Krikun, C. Lockwood, G. E. Akbas, and H. S. Taylor Differential Cell-Specific Modulation of HOXA10 by Estrogen and Specificity Protein 1 Response Elements J. Clin. Endocrinol. Metab., May 1, 2007; 92(5): 1920 - 1926. [Abstract] [Full Text] [PDF]
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 L. Mauro, S. Catalano, G. Bossi, M. Pellegrino, I. Barone, S. Morales, C. Giordano, V. Bartella, I. Casaburi, and S. Ando Evidences that Leptin Up-regulates E-Cadherin Expression in Breast Cancer: Effects on Tumor Growth and Progression Cancer Res., April 1, 2007; 67(7): 3412 - 3421. [Abstract] [Full Text] [PDF]
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 M. A. Stoner, S. S. Auerbach, S. M. Zamule, S. C. Strom, and C. J. Omiecinski Transactivation of a DR-1 PPRE by a human constitutive androstane receptor variant expressed from internal protein translation start sites Nucleic Acids Res., April 1, 2007; 35(7): 2177 - 2190. [Abstract] [Full Text] [PDF]
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 P. Galluzzo, F. Caiazza, S. Moreno, and M. Marino Role of ER{beta} palmitoylation in the inhibition of human colon cancer cell proliferation Endocr. Relat. Cancer, March 1, 2007; 14(1): 153 - 167. [Abstract] [Full Text] [PDF]
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D. G. Monroe, F. J. Secreto, J. R. Hawse, M. Subramaniam, S. Khosla, and T. C. Spelsberg Estrogen Receptor Isoform-specific Regulation of the Retinoblastoma-binding Protein 1 (RBBP1) Gene: ROLES OF AF1 AND ENHANCER ELEMENTS J. Biol. Chem., September 29, 2006; 281(39): 28596 - 28604. [Abstract] [Full Text] [PDF]
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J. S. Carroll and M. Brown Estrogen Receptor Target Gene: An Evolving Concept Mol. Endocrinol., August 1, 2006; 20(8): 1707 - 1714. [Abstract] [Full Text] [PDF]
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K. J. Higgins, S. Liu, M. Abdelrahim, K. Yoon, K. Vanderlaag, W. Porter, R. P. Metz, and S. Safe Vascular Endothelial Growth Factor Receptor-2 Expression Is Induced by 17{beta}-Estradiol in ZR-75 Breast Cancer Cells by Estrogen Receptor {alpha}/Sp Proteins Endocrinology, July 1, 2006; 147(7): 3285 - 3295. [Abstract] [Full Text] [PDF]
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F. Stossi, V. S. Likhite, J. A. Katzenellenbogen, and B. S. Katzenellenbogen Estrogen-occupied Estrogen Receptor Represses Cyclin G2 Gene Expression and Recruits a Repressor Complex at the Cyclin G2 Promoter J. Biol. Chem., June 16, 2006; 281(24): 16272 - 16278. [Abstract] [Full Text] [PDF]
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 P. Haeger, M. E. Andres, M. I. Forray, C. Daza, S. Araneda, and K. Gysling Estrogen receptors alpha and beta differentially regulate the transcriptional activity of the Urocortin gene. J. Neurosci., May 3, 2006; 26(18): 4908 - 4916. [Abstract] [Full Text] [PDF]
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M L Panno, L Mauro, S Marsico, D Bellizzi, P Rizza, C Morelli, M Salerno, F Giordano, and S Ando' Evidence that the mouse insulin receptor substrate-1 belongs to the gene family on which the promoter is activated by estrogen receptor {alpha} through its interaction with Sp1 J. Mol. Endocrinol., February 1, 2006; 36(1): 91 - 105. [Abstract] [Full Text] [PDF]
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 J. Malakooti, R. Sandoval, V. C. Memark, P. K. Dudeja, and K. Ramaswamy Zinc finger transcription factor Egr-1 is involved in stimulation of NHE2 gene expression by phorbol 12-myristate 13-acetate Am J Physiol Gastrointest Liver Physiol, October 1, 2005; 289(4): G653 - G663. [Abstract] [Full Text] [PDF]
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V. Gburcik, N. Bot, M. Maggiolini, and D. Picard SPBP Is a Phosphoserine-Specific Repressor of Estrogen Receptor {alpha} Mol. Cell. Biol., May 1, 2005; 25(9): 3421 - 3430. [Abstract] [Full Text] [PDF]
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A. Droit, G. G Poirier, and J. M Hunter Experimental and bioinformatic approaches for interrogating protein-protein interactions to determine protein function J. Mol. Endocrinol., April 1, 2005; 34(2): 263 - 280. [Abstract] [Full Text] [PDF]
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K. Kim, R. Barhoumi, R. Burghardt, and S. Safe Analysis of Estrogen Receptor {alpha}-Sp1 Interactions in Breast Cancer Cells by Fluorescence Resonance Energy Transfer Mol. Endocrinol., April 1, 2005; 19(4): 843 - 854. [Abstract] [Full Text] [PDF]
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L. Bjornstrom and M. Sjoberg Mechanisms of Estrogen Receptor Signaling: Convergence of Genomic and Nongenomic Actions on Target Genes Mol. Endocrinol., April 1, 2005; 19(4): 833 - 842. [Abstract] [Full Text] [PDF]
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 N. Kanda and S. Watanabe 17{beta}-Estradiol enhances heparin-binding epidermal growth factor-like growth factor production in human keratinocytes Am J Physiol Cell Physiol, April 1, 2005; 288(4): C813 - C823. [Abstract] [Full Text] [PDF]
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 H. Yuan, A. Gong, and C. Y.F. Young Involvement of transcription factor Sp1 in quercetin-mediated inhibitory effect on the androgen receptor in human prostate cancer cells Carcinogenesis, April 1, 2005; 26(4): 793 - 801. [Abstract] [Full Text] [PDF]
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J. E. Lee, K. Kim, J. C. Sacchettini, C. V. Smith, and S. Safe DRIP150 Coactivation of Estrogen Receptor {alpha} in ZR-75 Breast Cancer Cells Is Independent of LXXLL Motifs J. Biol. Chem., March 11, 2005; 280(10): 8819 - 8830. [Abstract] [Full Text] [PDF]
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Y-L Zhao, W-D Han, Q Li, Y-M Mu, X-C Lu, L Yu, H-J Song, X Li, J-M Lu, and C-Y Pan Mechanism of transcriptional regulation of LRP16 gene expression by 17-{beta} estradiol in MCF-7 human breast cancer cells J. Mol. Endocrinol., February 1, 2005; 34(1): 77 - 89. [Abstract] [Full Text] [PDF]
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A. Bardin, F. Moll, R. Margueron, C. Delfour, M. L. Chu, T. Maudelonde, V. Cavailles, and P. Pujol Transcriptional and Posttranscriptional Regulation of Fibulin-1 by Estrogens Leads to Differential Induction of Messenger Ribonucleic Acid Variants in Ovarian and Breast Cancer Cells Endocrinology, February 1, 2005; 146(2): 760 - 768. [Abstract] [Full Text] [PDF]
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N. Fujita, M. Kajita, P. Taysavang, and P. A. Wade Hormonal Regulation of Metastasis-Associated Protein 3 Transcription in Breast Cancer Cells Mol. Endocrinol., December 1, 2004; 18(12): 2937 - 2949. [Abstract] [Full Text] [PDF]
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J. Hong, I. Samudio, S. Liu, M. Abdelrahim, and S. Safe Peroxisome Proliferator-Activated Receptor {gamma}-Dependent Activation of p21 in Panc-28 Pancreatic Cancer Cells Involves Sp1 and Sp4 Proteins Endocrinology, December 1, 2004; 145(12): 5774 - 5785. [Abstract] [Full Text] [PDF]
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J. Ahn, M. D. Gammon, R. M. Santella, M. M. Gaudet, J. A. Britton, S. L. Teitelbaum, M. B. Terry, A. I. Neugut, P. D. Josephy, and C. B. Ambrosone Myeloperoxidase Genotype, Fruit and Vegetable Consumption, and Breast Cancer Risk Cancer Res., October 15, 2004; 64(20): 7634 - 7639. [Abstract] [Full Text] [PDF]
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A. E. Damdimopoulos, A. Miranda-Vizuete, E. Treuter, J.-A. Gustafsson, and G. Spyrou An Alternative Splicing Variant of the Selenoprotein Thioredoxin Reductase Is a Modulator of Estrogen Signaling J. Biol. Chem., September 10, 2004; 279(37): 38721 - 38729. [Abstract] [Full Text] [PDF]
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A Bardin, N Boulle, G Lazennec, F Vignon, and P Pujol Loss of ER{beta} expression as a common step in estrogen-dependent tumor progression Endocr. Relat. Cancer, September 1, 2004; 11(3): 537 - 551. [Abstract] [Full Text] [PDF]
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R. O'Lone, M. C. Frith, E. K. Karlsson, and U. Hansen Genomic Targets of Nuclear Estrogen Receptors Mol. Endocrinol., August 1, 2004; 18(8): 1859 - 1875. [Abstract] [Full Text] [PDF]
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 R. Grummer, S.W. Hewitt, O. Traub, K.S. Korach, and E. Winterhager Different Regulatory Pathways of Endometrial Connexin Expression: Preimplantation Hormonal-Mediated Pathway Versus Embryo Implantation-Initiated Pathway Biol Reprod, July 1, 2004; 71(1): 273 - 281. [Abstract] [Full Text] [PDF]
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M. N. Dieudonne, M. C. Leneveu, Y. Giudicelli, and R. Pecquery Evidence for functional estrogen receptors {alpha} and {beta} in human adipose cells: regional specificities and regulation by estrogens Am J Physiol Cell Physiol, March 1, 2004; 286(3): C655 - C661. [Abstract] [Full Text] [PDF]
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A. Huang, D. Sun, Z. Wu, C. Yan, M. A. Carroll, H. Jiang, J. R. Falck, and G. Kaley Estrogen Elicits Cytochrome P450--Mediated Flow-Induced Dilation of Arterioles in NO Deficiency: Role of PI3K-Akt Phosphorylation in Genomic Regulation Circ. Res., February 6, 2004; 94(2): 245 - 252. [Abstract] [Full Text] [PDF]
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 J. Spychala, E. Lazarowski, A. Ostapkowicz, L. H. Ayscue, A. Jin, and B. S. Mitchell Role of Estrogen Receptor in the Regulation of Ecto-5'-Nucleotidase and Adenosine in Breast Cancer Clin. Cancer Res., January 15, 2004; 10(2): 708 - 717. [Abstract] [Full Text] [PDF]
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 M. E. Colosimo, S. Tran, and P. Sengupta The Divergent Orphan Nuclear Receptor ODR-7 Regulates Olfactory Neuron Gene Expression via Multiple Mechanisms in Caenorhabditis elegans Genetics, December 1, 2003; 165(4): 1779 - 1791. [Abstract] [Full Text] [PDF]
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D. C. Spink, B. H. Katz, M. M. Hussain, B. T. Pentecost, Z. Cao, and B. C. Spink Estrogen regulates Ah responsiveness in MCF-7 breast cancer cells Carcinogenesis, December 1, 2003; 24(12): 1941 - 1950. [Abstract] [Full Text] [PDF]
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S. L. Zheng, W. Zheng, B.-l. Chang, X.-O. Shu, Q. Cai, H. Yu, Q. Dai, J. Xu, and Y.-T. Gao Joint Effect of Estrogen Receptor {beta} Sequence Variants and Endogenous Estrogen Exposure on Breast Cancer Risk in Chinese Women Cancer Res., November 15, 2003; 63(22): 7624 - 7629. [Abstract] [Full Text] [PDF]
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D. Liu, Z. Zhang, W. Gladwell, and C. T. Teng Estrogen Stimulates Estrogen-Related Receptor {alpha} Gene Expression through Conserved Hormone Response Elements Endocrinology, November 1, 2003; 144(11): 4894 - 4904. [Abstract] [Full Text] [PDF]
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Y. Kinoshita and S. Chen Induction of Aromatase (CYP19) Expression in Breast Cancer Cells through a Nongenomic Action of Estrogen Receptor {alpha} Cancer Res., July 1, 2003; 63(13): 3546 - 3555. [Abstract] [Full Text] [PDF]
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S. Khan, M. Abdelrahim, I. Samudio, and S. Safe Estrogen Receptor/Sp1 Complexes Are Required for Induction of cad Gene Expression by 17{beta}-Estradiol in Breast Cancer Cells Endocrinology, June 1, 2003; 144(6): 2325 - 2335. [Abstract] [Full Text] [PDF]
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S. Harendza, D. H. Lovett, U. Panzer, Z. Lukacs, P. Kuhnl, and R. A. K. Stahl Linked Common Polymorphisms in the Gelatinase A Promoter Are Associated with Diminished Transcriptional Response to Estrogen and Genetic Fitness J. Biol. Chem., May 30, 2003; 278(23): 20490 - 20499. [Abstract] [Full Text] [PDF]
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S. Ngwenya and S. Safe Cell Context-Dependent Differences in the Induction of E2F-1 Gene Expression by 17{beta}-Estradiol in MCF-7 and ZR-75 Cells Endocrinology, May 1, 2003; 144(5): 1675 - 1685. [Abstract] [Full Text] [PDF]
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K. Kim, N. Thu, B. Saville, and S. Safe Domains of Estrogen Receptor {alpha} (ER{alpha}) Required for ER{alpha}/Sp1-Mediated Activation of GC-Rich Promoters by Estrogens and Antiestrogens in Breast Cancer Cells Mol. Endocrinol., May 1, 2003; 17(5): 804 - 817. [Abstract] [Full Text] [PDF]
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M. Wormke, M. Stoner, B. Saville, K. Walker, M. Abdelrahim, R. Burghardt, and S. Safe The Aryl Hydrocarbon Receptor Mediates Degradation of Estrogen Receptor {alpha} through Activation of Proteasomes Mol. Cell. Biol., March 15, 2003; 23(6): 1843 - 1855. [Abstract] [Full Text] [PDF]
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V. Sriraman, S. C. Sharma, and J. S. Richards Transactivation of the Progesterone Receptor Gene in Granulosa Cells: Evidence that Sp1/Sp3 Binding Sites in the Proximal Promoter Play a Key Role in Luteinizing Hormone Inducibility Mol. Endocrinol., March 1, 2003; 17(3): 436 - 449. [Abstract] [Full Text] [PDF]
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A. Pedram, M. Razandi, M. Aitkenhead, C. C. W. Hughes, and E. R. Levin Integration of the Non-genomic and Genomic Actions of Estrogen. MEMBRANE-INITIATED SIGNALING BY STEROID TO TRANSCRIPTION AND CELL BIOLOGY J. Biol. Chem., December 20, 2002; 277(52): 50768 - 50775. [Abstract] [Full Text] [PDF]
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L. Bjornstrom and M. Sjoberg Mutations in the Estrogen Receptor DNA-binding Domain Discriminate between the Classical Mechanism of Action and Cross-talk with Stat5b and Activating Protein 1 (AP-1) J. Biol. Chem., December 6, 2002; 277(50): 48479 - 48483. [Abstract] [Full Text] [PDF]
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M. Jakacka, M. Ito, F. Martinson, T. Ishikawa, E. J. Lee, and J. L. Jameson An Estrogen Receptor (ER){alpha} Deoxyribonucleic Acid-Binding Domain Knock-In Mutation Provides Evidence for Nonclassical ER Pathway Signaling in Vivo Mol. Endocrinol., October 1, 2002; 16(10): 2188 - 2201. [Abstract] [Full Text] [PDF]
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 L. Molero, M. Garcia-Duran, J. Diaz-Recasens, L. Rico, S. Casado, and A. Lopez-Farre Expression of estrogen receptor subtypes and neuronal nitric oxide synthase in neutrophils from women and men: Regulation by estrogen Cardiovasc Res, October 1, 2002; 56(1): 43 - 51. [Abstract] [Full Text] [PDF]
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M. Abdelrahim, I. Samudio, R. Smith III, R. Burghardt, and S. Safe Small Inhibitory RNA Duplexes for Sp1 mRNA Block Basal and Estrogen-induced Gene Expression and Cell Cycle Progression in MCF-7 Breast Cancer Cells J. Biol. Chem., August 2, 2002; 277(32): 28815 - 28822. [Abstract] [Full Text] [PDF]
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C. Lethimonier, G. Flouriot, O. Kah, and B. Ducouret The Glucocorticoid Receptor Represses the Positive Autoregulation of the Trout Estrogen Receptor Gene by Preventing the Enhancer Effect of a C/EBP{beta}-Like Protein Endocrinology, August 1, 2002; 143(8): 2961 - 2974. [Abstract] [Full Text] [PDF]
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C. Qin, T. Nguyen, J. Stewart, I. Samudio, R. Burghardt, and S. Safe Estrogen Up-Regulation of p53 Gene Expression in MCF-7 Breast Cancer Cells Is Mediated by Calmodulin Kinase IV-Dependent Activation of a Nuclear Factor {kappa}B/CCAAT-Binding Transcription Factor-1 Complex Mol. Endocrinol., August 1, 2002; 16(8): 1793 - 1809. [Abstract] [Full Text] [PDF]
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T. R. Ediger, S.-E. Park, and B. S. Katzenellenbogen Estrogen Receptor Inducibility of the Human Na+/H+ Exchanger Regulatory Factor/Ezrin-Radixin-Moesin Binding Protein 50 (NHE-RF/EBP50) Gene Involving Multiple Half-Estrogen Response Elements Mol. Endocrinol., August 1, 2002; 16(8): 1828 - 1839. [Abstract] [Full Text] [PDF]
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P. Meisel Cancer, genes and gender Carcinogenesis, June 1, 2002; 23(6): 1087 - 1088. [Full Text] [PDF]
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 E. R. Levin Genome and Hormones: Gender Differences in Physiology: Invited Review: Cell localization, physiology, and nongenomic actions of estrogen receptors J Appl Physiol, October 1, 2001; 91(4): 1860 - 1867. [Abstract] [Full Text] [PDF]
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P. G. V. Martini and B. S. Katzenellenbogen Regulation of Prothymosin {alpha} Gene Expression by Estrogen in Estrogen Receptor-Containing Breast Cancer Cells via Upstream Half-Palindromic Estrogen Response Element Motifs Endocrinology, August 1, 2001; 142(8): 3493 - 3501. [Abstract] [Full Text] [PDF]
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C. M. Klinge Estrogen receptor interaction with estrogen response elements Nucleic Acids Res., July 15, 2001; 29(14): 2905 - 2919. [Abstract] [Full Text] [PDF]
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C. Li, M. R. Briggs, T. E. Ahlborn, F. B. Kraemer, and J. Liu Requirement of Sp1 and Estrogen Receptor {{alpha}} Interaction in 17{beta}-Estradiol-Mediated Transcriptional Activation of the Low Density Lipoprotein Receptor Gene Expression Endocrinology, April 1, 2001; 142(4): 1546 - 1553. [Abstract] [Full Text]
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B. Morin, G. R. Woodcock, L. A. Nichols, and L. J. Holland Heterodimerization between the Glucocorticoid Receptor and the Unrelated DNA-Binding Protein, Xenopus Glucocorticoid Receptor Accessory Factor Mol. Endocrinol., March 1, 2001; 15(3): 458 - 466. [Abstract] [Full Text]
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I. Samudio, C. Vyhlidal, F. Wang, M. Stoner, I. Chen, M. Kladde, R. Barhoumi, R. Burghardt, and S. Safe Transcriptional Activation of Deoxyribonucleic Acid Polymerase {{alpha}} Gene Expression in MCF-7 Cells by 17{{beta}}-Estradiol Endocrinology, March 1, 2001; 142(3): 1000 - 1008. [Abstract] [Full Text] [PDF]
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J. E. Compston Sex Steroids and Bone Physiol Rev, January 1, 2001; 81(1): 419 - 447. [Abstract] [Full Text] [PDF]
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M. Razandi, A. Pedram, and E. R. Levin Plasma Membrane Estrogen Receptors Signal to Antiapoptosis in Breast Cancer Mol. Endocrinol., September 1, 2000; 14(9): 1434 - 1447. [Abstract] [Full Text]
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O. Loudig, C. Babichuk, J. White, S. Abu-Abed, C. Mueller, and M. Petkovich Cytochrome P450RAI(CYP26) Promoter: A Distinct Composite Retinoic Acid Response Element Underlies the Complex Regulation of Retinoic Acid Metabolism Mol. Endocrinol., September 1, 2000; 14(9): 1483 - 1497. [Abstract] [Full Text]
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T. Sugawara, M. Saito, and S. Fujimoto Sp1 and SF-1 Interact and Cooperate in the Regulation of Human Steroidogenic Acute Regulatory Protein Gene Expression Endocrinology, August 1, 2000; 141(8): 2895 - 2903. [Abstract] [Full Text] [PDF]
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L. N. Petz and A. M. Nardulli Sp1 Binding Sites and An Estrogen Response Element Half-Site Are Involved in Regulation of the Human Progesterone Receptor A Promoter Mol. Endocrinol., July 1, 2000; 14(7): 972 - 985. [Abstract] [Full Text]
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W. Xie, R. Duan, I. Chen, I. Samudio, and S. Safe Transcriptional Activation of Thymidylate Synthase by 17{beta}-Estradiol in MCF-7 Human Breast Cancer Cells Endocrinology, July 1, 2000; 141(7): 2439 - 2449. [Abstract] [Full Text] [PDF]
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S. Misiti, S. Nanni, G. Fontemaggi, Y.-S. Cong, J. Wen, H. W. Hirte, G. Piaggio, A. Sacchi, A. Pontecorvi, S. Bacchetti, et al. Induction of hTERT Expression and Telomerase Activity by Estrogens in Human Ovary Epithelium Cells Mol. Cell. Biol., June 1, 2000; 20(11): 3764 - 3771. [Abstract] [Full Text]
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L. Salvatori, L. Ravenna, M. P. Felli, M. R. Cardillo, M. A. Russo, L. Frati, A. Gulino, and E. Petrangeli Identification of an Estrogen-Mediated Deoxyribonucleic Acid-Binding Independent Transactivation Pathway on the Epidermal Growth Factor Receptor Gene Promoter Endocrinology, June 1, 2000; 141(6): 2266 - 2274. [Abstract] [Full Text] [PDF]
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C. Lethimonier, G. Flouriot, Y. Valotaire, O. Kah, and B. Ducouret Transcriptional Interference Between Glucocorticoid Receptor and Estradiol Receptor Mediates the Inhibitory Effect of Cortisol on Fish Vitellogenesis Biol Reprod, June 1, 2000; 62(6): 1763 - 1771. [Abstract] [Full Text]
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A. van den Wijngaard, W. R. Mulder, R. Dijkema, C. J .C. Boersma, S. Mosselman, E. J. J. van Zoelen, and W. Olijve Antiestrogens Specifically Up-Regulate Bone Morphogenetic Protein-4 Promoter Activity in Human Osteoblastic Cells Mol. Endocrinol., May 1, 2000; 14(5): 623 - 633. [Abstract] [Full Text]
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S. Oesterreich, Q. Zhang, T. Hopp, S. A. W. Fuqua, M. Michaelis, H. H. Zhao, J. R. Davie, C. K. Osborne, and A. V. Lee Tamoxifen-Bound Estrogen Receptor (ER) Strongly Interacts with the Nuclear Matrix Protein HET/SAF-B, a Novel Inhibitor of ER-Mediated Transactivation Mol. Endocrinol., March 1, 2000; 14(3): 369 - 381. [Abstract] [Full Text]
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B. Saville, M. Wormke, F. Wang, T. Nguyen, E. Enmark, G. Kuiper, J.-A. Gustafsson, and S. Safe Ligand-, Cell-, and Estrogen Receptor Subtype (alpha /beta )-dependent Activation at GC-rich (Sp1) Promoter Elements J. Biol. Chem., February 25, 2000; 275(8): 5379 - 5387. [Abstract] [Full Text] [PDF]
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H. Poukka, P. Aarnisalo, H. Santti, O. A. Janne, and J. J. Palvimo Coregulator Small Nuclear RING Finger Protein (SNURF) Enhances Sp1- and Steroid Receptor-mediated Transcription by Different Mechanisms J. Biol. Chem., January 7, 2000; 275(1): 571 - 579. [Abstract] [Full Text] [PDF]
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S. Kyo, M. Takakura, T. Kanaya, W. Zhuo, K. Fujimoto, Y. Nishio, A. Orimo, and M. Inoue Estrogen Activates Telomerase Cancer Res., December 1, 1999; 59(23): 5917 - 5921. [Abstract] [Full Text] [PDF]
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P. S. Jones, E. Parrott, and I. N. H. White Activation of Transcription by Estrogen Receptor alpha and beta Is Cell Type- and Promoter-dependent J. Biol. Chem., November 5, 1999; 274(45): 32008 - 32014. [Abstract] [Full Text] [PDF]
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L. Dong, W. Wang, F. Wang, M. Stoner, J. C. Reed, M. Harigai, I. Samudio, M. P. Kladde, C. Vyhlidal, and S. Safe Mechanisms of Transcriptional Activation of bcl-2 Gene Expression by 17beta -Estradiol in Breast Cancer Cells J. Biol. Chem., November 5, 1999; 274(45): 32099 - 32107. [Abstract] [Full Text] [PDF]
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R. Duan, W. Porter, I. Samudio, C. Vyhlidal, M. Kladde, and S. Safe Transcriptional Activation of c-fos Protooncogene by 17{beta}-Estradiol: Mechanism of Aryl Hydrocarbon Receptor-Mediated Inhibition Mol. Endocrinol., September 1, 1999; 13(9): 1511 - 1521. [Abstract] [Full Text]
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S. C. Sharma, J. W. Clemens, M. D. Pisarska, and J. S. Richards Expression and Function of Estrogen Receptor Subtypes in Granulosa Cells: Regulation by Estradiol and Forskolin Endocrinology, September 1, 1999; 140(9): 4320 - 4334. [Abstract] [Full Text]
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W. Wang, L. Dong, B. Saville, and S. Safe Transcriptional Activation of E2F1 Gene Expression by 17{beta}-Estradiol in MCF-7 Cells Is Regulated by NF-Y-Sp1/Estrogen Receptor Interactions Mol. Endocrinol., August 1, 1999; 13(8): 1373 - 1387. [Abstract] [Full Text]
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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]
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D. S. Hunter, L. C. Hodges, P. M. Vonier, R. Fuchs-Young, M. M. Gottardis, and C. L. Walker Estrogen Receptor Activation via Activation Function 2 Predicts Agonism of Xenoestrogens in Normal and Neoplastic Cells of the Uterine Myometrium Cancer Res., July 1, 1999; 59(13): 3090 - 3099. [Abstract] [Full Text] [PDF]
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C. Qin, P. Singh, and S. Safe Transcriptional Activation of Insulin-Like Growth Factor-Binding Protein-4 by 17{beta}-Estradiol in MCF-7 Cells: Role of Estrogen Receptor-Sp1 Complexes Endocrinology, June 1, 1999; 140(6): 2501 - 2508. [Abstract] [Full Text]
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F. Machinal, M.-N. Dieudonne, M.-C. Leneveu, R. Pecquery, and Y. Giudicelli In Vivo and in Vitro ob Gene Expression and Leptin Secretion in Rat Adipocytes: Evidence for a Regional Specific Regulation by Sex Steroid Hormones Endocrinology, April 1, 1999; 140(4): 1567 - 1574. [Abstract] [Full Text]
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