Progesterone Stimulates Adipocyte Determination and
Differentiation 1/Sterol Regulatory Element-binding Protein 1c Gene
Expression
POTENTIAL MECHANISM FOR THE LIPOGENIC EFFECT OF PROGESTERONE IN
ADIPOSE TISSUE*
Danièle
Lacasa
,
Xavier
Le Liepvre,
Pascal
Ferre, and
Isabelle
Dugail§
From the
Laboratoire de Biochimie et Biologie
Moléculaire, Faculté de Médecine Paris Ouest,
Université René Descartes, 75270 Paris, France and
INSERM U 465, Nutrition, Métabolisme, Obésité, 15 Rue
de l'École de Médecine, 75006 Paris, France
Received for publication, September 19, 2000, and in revised form, December 14, 2000
 |
ABSTRACT |
Fatty acid synthase (FAS), a
nutritionally regulated lipogenic enzyme, is transcriptionally
controlled by ADD1/SREBP1c (adipocyte determination and differentiation
1/sterol regulatory element-binding protein 1c), through
insulin-mediated stimulation of ADD1/SREBP1c expression.
Progesterone exerts lipogenic effects on adipocytes, and FAS is highly
induced in breast tumor cell lines upon progesterone treatment. We show
here that progesterone up-regulates ADD1/SREBP1c expression in the MCF7
breast cancer cell line and the primary cultured preadipocyte from rat
parametrial adipose tissue. In MCF7, progesterone induced ADD1/SREBP1c
and Metallothionein II (a well known progesterone-regulated gene)
mRNAs, with comparable potency. In preadipocytes, progesterone
increased ADD1/SREBP1c mRNA dose-dependently, but not
SREBP1a or SREBP2. Run-on experiments demonstrated that progesterone
action on ADD1/SREBP1c was primarily at the transcriptional level. The
membrane-bound and mature nuclear forms of ADD1/SREBP1 protein
accumulated in preadipocytes cultured with progesterone, and FAS
induction could be abolished by adenovirus-mediated overexpression of a
dominant negative form of ADD1/SREBP1 in these cells. Finally, in the
presence of insulin, progesterone was unable to up-regulate
ADD1/SREBP1c mRNA in preadipocytes, whereas its effect was restored
after 24 h of insulin deprivation. Together these results
demonstrate that ADD1/SREBP1c is controlled by progesterone, which,
like insulin, acts by increasing ADD1/SREBP1c gene transcription. This
provides a potential mechanism for the lipogenic actions of
progesterone on adipose tissue.
 |
INTRODUCTION |
Fatty acid synthase
(FAS)1 is a multifunctional
enzyme that catalyzes all the steps in the synthesis of long chain
fatty acids from malonyl CoA. As a key lipogenic enzyme, FAS is
expressed mainly in liver and adipose tissue, where it turns dietary
carbohydrates to fat. In these tissues, the transcription of the FAS
gene is under nutritional control, leading to commensurately regulated activity of the enzyme. Briefly, feeding a high carbohydrate diet induces, whereas fasting or consuming a high fat diet decreases, FAS
gene expression. Insulin (1), glucose (2), fatty acids (3-5), and cAMP
(6) are direct effectors of the nutritional regulation of FAS, exerting
coordinated effects on FAS gene transcription at the promoter level.
High levels of FAS expression are also found in some tumor cells of
breast cancer (7-9) and derived cell lines, where it is associated
with a worsened prognosis (10). cDNA for FAS has been cloned
initially as a progestin-responsive mRNA by differential screening
of the MCF7 breast cancer cell line (11), and further studies have
established that FAS expression was induced by progestins in the normal
mammary gland also (12). The mechanism of FAS induction by progestins
relies primarily on transcriptional activation, as shown by run-on
studies (13). However, the direct implication of the progesterone
receptor in the FAS gene-stimulated transcription has not been clearly established.
Recently, significant progress has been made in the elucidation of the
mechanisms of FAS gene regulation. Particularly, the role of a key
transcription factor, ADD1/SREBP1c (adipocyte determination and
differentiation 1/sterol regulatory element-binding protein 1c), has
been uncovered. ADD1/SREBP1c, a member of the basic helix loop helix
family of transcription factors (14, 15), has been identified as a
potent activator of the FAS promoter in cultured cells (16) and in
transgenic mice (17). The ability of ADD1/SREBP1c to transactivate the
FAS gene seems to be physiologically relevant, because ADD1/SREBP1c is
induced by insulin in primary hepatocytes (18) and adipose cell lines
(19), down-regulated by cAMP and glucagon in hepatocytes (18), and
nutritionally regulated in the liver of mice (20). For these reasons,
it has been proposed that ADD1/SREBP1c is the mediator of insulin
action on FAS gene expression (21).
In the light of these new insights, the present study was designed to
investigate the mechanisms of FAS gene regulation by progesterone. We
found that progesterone is able to stimulate ADD1/SREBP1c expression,
making it likely that progesterone-induced stimulation of FAS
expression is exerted through activation of the same transcription
factor as insulin, ADD1/SREBP1c.
 |
EXPERIMENTAL PROCEDURES |
Materials--
Fetal bovine serum was obtained from Life
Technologies, Inc. Phenol red-free DMEM containing 4.5 g/liter
glucose and DMEM, Ham's F12 (50:50 mix) were obtained from Sigma.
Animals--
Procedures with experimental animals were
authorized and followed the guidelines of the Ministry of Agriculture
(France) (authorization 006614). Female Harlan Sprague-Dawley
rats (125-150 g) were killed by decapitation, and parametrial fat pads
were removed aseptically.
Cell Culture and Adenovirus-mediated Gene Transfer--
Cell
preparation and culture were performed as described in Ref. 22.
Briefly, preadipocytes obtained from the stroma-vascular fraction of
adipose tissue by collagenase digestion were plated at a density of
1-2 × 104 cells/cm2 in 8% fetal bovine
serum/DMEM. After 12 h, cultures were washed and fed with 8%
fetal bovine serum/DMEM. Medium was changed every other day. At
confluence (3 days post-plating), cells were allowed to differentiate
in DMEM/Ham's F12 containing 5 µg/ml insulin, 10 µg/ml
transferrin, and 200 pM T3 (ITT medium) in the absence of
serum, as described in Ref. 22. Early differentiating preadipocytes (day 2 post-confluence) were treated with progesterone in serum-free medium for 24 h unless otherwise stated. Progesterone treatment was provided to cell dishes as an ethanol solution. An equivalent volume of ethanol alone (never exceeding 0.1% v/v) was added in untreated controls.
In some experiments, cells in serum-free medium were infected (100 plaque-forming units/cell) with an adenovirus encoding a dominant
negative form of ADD1 under the control of the cytomegalovirus promoter
(ad-DN) or with a control empty virus (ad-null) as described (23).
16 h post-infection, the medium was changed, and progesterone was
added or not for the next 24 h. MCF7 cells were obtained from ATCC
(Manassas, Va) and cultured as recommended by the supplier. Progesterone treatment was performed in serum-free medium 1 day after confluence.
RNA Isolation and Northern Blot Analysis--
Total RNA was
isolated from 3-5 culture dishes (90 mm) by the guanidium thiocyanate
method, as described in Ref. 24. RNAs were then separated on
formaldehyde-agarose gels and transferred onto nylon membranes (Hybond
N+, Amersham Pharmacia Biotech). Hybridization was as described
previously (23), and blots were washed in 0.1% SSC, 0.1% SDS at
60 °C. Hybridization probes were as follows. ADD1/SREBP1c
probe was a rat ADD1 cDNA fragment encompassing the first 403 amino
acids of the ADD1 protein cloned in pSVSPORT1 (provided by B. Spiegelman, Boston, MA). The SREBP1a probe was a polymerase chain
reaction fragment described previously (18). The plasmid encoding
full-length MTII is described in Ref. 25 and was a kind gift of Dr. P. Hainaut (International Agency for Research on Cancer, Lyon, France). An
RNA probe for 18 S was used for normalization of the results.
Run-on Experiments--
The effect of progesterone on
gene transcription was assessed by run-on experiments as described in
Ref. 26.
Western Blot Analysis--
Nuclear extracts and crude membranes
were prepared from cultured preadipocytes as described previously (27),
separated on 8% polyacrylamide-SDS gels, and electrotransferred to
nylon membranes (Amersham Pharmacia Biotech). SREBP1 was probed using a
5 µg/ml dilution of the polyclonal antibody IgG-2A4 (ATCC). A
C/EBP
antibody (SC130, Santa Cruz Biotechnology) was also
used as a control for the specificity of progesterone effect. The blots
were revealed using the ECL system (Pierce), as described by the manufacturer.
Statistical Analysis of the Results--
The effect of
progesterone was evaluated by Dunnet's Post test.
 |
RESULTS |
The stimulatory effect of progesterone on FAS gene expression was
originally described in MCF7, a breast carcinoma cell line. As a first
step, the ability of progesterone to induce FAS mRNA in cultured
preadipocytes was compared with the MCF7 cell system. A model of
cultured preadipocytes isolated from female parametrial adipose tissue
was chosen because of the presence of well characterized progesterone
receptors on these cells (28) and the induction of FAS activity upon
differentiation. MCF7 cells or early differentiating preadipocytes (day 2 post-confluence) were incubated for 24 h in
serum-free medium in the presence of increasing concentrations of
progesterone, and FAS mRNA levels were assessed by Northern blot
analysis. Fig. 1 shows that progesterone
dose-dependently increases FAS mRNA in both MCF7
cells and preadipocytes. We have also observed that induction of
FAS mRNA by progesterone in preadipocytes was accompanied by a
significant increase in lipogenic activity, as assessed by the
conversion of [U-14C]glucose into lipids (data not
shown). This establishes that the system of primary cultured
preadipocytes behaves as the MCF7 cell line and is suitable to study
progesterone regulation of FAS activity.

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Fig. 1.
Northern blot analysis of FAS mRNA in
MCF7 cells (left) or in primary
cultured preadipocytes (right) treated for 24 h
in the presence of increasing concentrations of progesterone, as
described under "Experimental Procedures." For MCF7
cells, a typical dose-response curve obtained from two
experiments is shown. For preadipocytes, each bar represents
the mean value ± S.E. from three independent cultures. The effect
of progesterone is statistically significant at the p < 0.05 level by Dunnet's Post test. C,
control.
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|
In the light of the importance of ADD1/SREBP1c in insulin-mediated
regulation of FAS gene expression, the effect of progesterone on SREBPs
was next investigated. Fig. 2 shows that
using an ADD1/SREBP1 cDNA probe, a dose-dependent
induction of progesterone on ADD1/SREBP1 mRNA levels could be
detected in preadipocytes. The EC50 for the progesterone
effect was 90 nM, in good relation to that observed for the
induction of FAS gene expression (220 nM) by the hormone (see Fig. 1). The ADD1/SREBP1 gene can be transcribed from two alternate promoters, generating two different (ADD1/SREBP1c and SREBP1a) transcripts of approximately the same size. Because the ADD1/SREBP1 probe used did not distinguish between the 1c and the 1a
transcript, we also used a probe specific for SREBP1a to examine the
effect of progesterone. We show in Fig. 2 that this 1a probe generated
very weak hybridization signals that required a long time exposure and
did not reveal any effect of progesterone. This suggests that in
preadipocytes, the expression of the 1a isoform is low and unaffected
by hormone treatment. Thus we concluded that the progesterone-induced
ADD1/SREBP1 mRNA represented mainly the ADD1/SREBP1c transcript.
For these reasons, the signals generated by the ADD1/SREBP1 probe were
identified as ADD1/SREBP1c mRNA in quantitative analysis in Fig.
2B. Because it has been demonstrated that progesterone was
able to modify cholesterol trafficking at the plasma membrane, by
virtue of its amphiphile properties, we also probed the blots for the
mRNA encoding SREBP2, the cholesterol-sensitive isoform of SREBP
that is derived from an independent gene. No hybridization signal for
SREBP2 could be detected by Northern blot analysis, and no induction of
SREBP2 mRNA could be seen upon progesterone treatment (data not
shown). These data indicate that progesterone selectively stimulates
the expression of the ADD1/SREBP1c mRNA, but not SREBP1a or SREBP2.
The time course of the induction of ADD1/SREBP1c by progesterone was
also examined (Fig. 2C). ADD1/SREBP1c mRNA levels
increased between 0 and 24 h in control cells treated with ethanol
only, reflecting the differentiation-dependent expression of ADD1/SREBP1c during adipose conversion. In progesterone-treated cells, ADD1/SREBP1c induction was more marked than in controls, indicating an effect of progesterone beginning after 6 h of
treatment, sustained for at least a 24-h period.

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Fig. 2.
Effect of progesterone on ADD1/SREBP1
mRNA levels in primary cultured preadipocytes. A shows
Northern blots from primary cultured preadipocytes treated for 24 h with progesterone. B presents quantification of the
dose-response curves for ADD1/SREBP1c mRNA. Values were obtained
from at least four independent cultures. The effect of progesterone is
statistically significant at the p < 0.05 level by
Dunnet's Post test. C, control. C shows
time course experiments of the effect of progesterone on ADD1/SREBP1c
mRNA. Progesterone (1 µM) was added as an ethanol
solution, and ethanol only was added in control cells. A typical
experiment of two is shown.
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Among well characterized progesterone target genes is the
Metallothionein IIA (MT-IIA) (29). Thus, we next compared the effects
of progesterone on MT-IIA and ADD1/SREBP1c mRNA levels in the MCF7
cell line. Only background levels of hybridization could be detected in
MCF7 cells with the SREBP1a-specific probe (data not shown), suggesting
that, as in preadipocytes, the signals obtained with the ADD1/SREBP1
probe were generated by ADD1/SREBP1c. Fig.
3 shows that ADD1/SREBP1c mRNA is
expressed, albeit at low levels, in MCF7 cells in the absence of
progesterone. This basal level of ADD1/SREBP1c mRNA can be
dose-dependently induced by progesterone, in a very similar
manner to the induction observed for the mRNA of MT-IIA, a well
known progesterone-responsive gene. EC5O values were 110 and 70 nM for ADD1/SREBP1c and MT-IIA mRNAs, respectively. The time course of progesterone induction of ADD1/SREBP1c mRNA was also very similar to that reported in preadipocytes, with
a stimulatory effect detectable after 6 h (data not shown). Thus
these results demonstrate that ADD1/SREBP1c, like MT-IIA, is a
progesterone-inducible gene. To further investigate the mechanism by
which ADD1/SREBP1c mRNA concentrations are increased by
progesterone, we measured gene transcription rates in run-on
experiments. Fig. 4 shows that the
ADD1/SREBP1 transcription rate is increased by progesterone treatment
in both MCF7 cells and preadipocytes. Moreover, the amplitude of
the progesterone effect on transcription (6.5-fold in MCF7 cells
and 3.2-fold in preadipocytes) closely parallels that observed for
steady state mRNA levels (9.7-fold in MCF7 cells and
3.1-fold in preadipocytes). Thus progesterone acts primarily at the
transcriptional level in the regulation of ADD1/SREBP1c.

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Fig. 3.
Effect of progesterone on ADD1/SREBP1c and
MT-IIA mRNA levels in MCF7 cells. MCF7 cells were treated with
increasing concentrations of progesterone for 24 h as described
under "Experimental Procedures." Representative blots are shown on
the left, and quantification of the signals (normalized to
18 S) is shown on the right. C,
control.
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Fig. 4.
Effect of progesterone on ADD1/SREBP1c
transcription rate. MCF7 cells (upper panel)
and preadipocytes (lower panel) were treated for 24 h
with 10 µM progesterone (Pg) as
described under "Experimental Procedures," and nuclei were
prepared. Labeled RNAs were hybridized to 10 µg of dot-blotted
plasmids. The control plasmid was the pSVsport1 vector in which
ADD1/SREBP1 is cloned. The glyceraldehyde-3-phosphate dehydrogenase
(GAPDH) plasmid (full-length rat glyceraldehyde-3-phosphate
dehydrogenase cDNA in pUC9) was used as a negative control.
Autoradiograms show representative results obtained with two
independent preparations of nuclei. Quantification of the blots is
shown on the right.
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To establish the link between the induction of ADD1/SREBP1c mRNA
expression by progesterone and the induction of the FAS gene, an
ADD1/SREBP1c target, we first investigated whether progesterone was
able to increase the amount of the ADD1/SREBP1 protein in the cells.
Crude membranes and nuclear extracts of preadipocytes were prepared and
probed with a monoclonal anti-SREBP1 antibody in Western blots. As
shown in Fig. 5A, cells
treated with progesterone for 24 h showed increased levels of the
precursor form in membrane fractions and higher contents of the active
nuclear (cleaved) form of the ADD1/SREBP1 protein. As a control for the
specificity of the progesterone effect, nuclear extracts were also
probed with a C/EBP
antibody, which did not reveal any change in
C/EBP
protein content. Collectively, these data demonstrate that
progesterone, by increasing transcription rates of the gene, raises the
levels of ADD1/SREBP1 mRNA in cells, which in turn leads to the
accumulation of the membrane-bound precursor and mature nuclear forms
of the ADD1/SREBP1 protein.

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Fig. 5.
The effect of progesterone on the FAS gene is
mediated through ADD1/SREBP1. A, Western blot analysis of
nuclear extracts and crude membrane fractions from preadipocytes
treated or not with progesterone. Blots were probed with a monoclonal
SREBP1 antibody. A C/EBP antibody was used as a control for the
specificity of the progesterone effect in nuclear extracts. This
antibody recognized several C/EBP isoforms, which were not resolved
on a 8% polyacrylamide-SDS gel. Similar results were obtained in three
independent preparations of nuclear extracts and membranes.
kD, kilodaltons. B, effect of adenovirus-mediated
overexpression of a dominant negative form of ADD1/SREBP1c on
progesterone (Prog)-induced FAS gene expression.
Preadipocytes from parametrial adipose tissue from females (1 day after
confluence) were infected with 100 plaque-forming units/cell of an
adenovirus encoding or not (ad-null) a dominant negative form of ADD1
(ad-DN). 16 h post-infection, the serum-free medium was
supplemented or not with 10 6 M
progesterone. Total RNA was extracted from five pooled dishes 24 h
later. A representative blot of two independent experiments is
shown.
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To definitely establish the link between the induction of the nuclear
mature form of ADD1/SREBP1 and FAS gene expression, we examined whether
overexpression of a dominant negative form of ADD1/SREBP1c in
progesterone-treated cells was able to block FAS gene induction.
Preadipocytes were thus infected with an adenovirus overexpressing a
dominant negative mutant for ADD1, described in Ref. 23, or a control
null vector and then stimulated by progesterone. The dominant negative
form of ADD1 has been demonstrated to sequester endogenous ADD1/SREBP1c
in cells, by dimerizing with the wild type protein and preventing DNA
binding (30). Results in Fig. 5B show that the 4-fold
induction of FAS expression by progesterone that occurred in cells
infected by the null vector was abolished when cells were infected by
the adenovirus encoding the dominant negative form of ADD1. This
demonstrates that ADD1/SREBP1 is required for progesterone-induced FAS
gene expression.
Finally, we investigated the relationship between insulin and
progesterone in the control of ADD1/SREBP1c expression. Preadipocytes, which are known to acquire insulin sensitivity upon differentiation, were induced to fully differentiate in the presence of insulin (day 7 post-confluence), and the ability of progesterone to induce ADD1/SREBP1c expression was tested. Fig.
6 shows that when cells differentiated in
the presence of insulin, the ability of progesterone to induce
ADD1/SREBP1c expression was abolished. The lack of progesterone effect
was not due to a general desensitizing effect of insulin to
progesterone, because MT-IIA gene expression still responded normally
in insulin-differentiated cells. In agreement, Fig. 6 also shows that
progesterone response was restored when cells were allowed to fully
differentiate in the presence of insulin and then deprived of the
hormone 24 h before progesterone treatment. Thus this experiment
shows that insulin and progesterone exert nonadditive effects on
ADD1/SREBP1c gene expression.

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Fig. 6.
Interaction between insulin and progesterone
on ADD1/SREBP1c, MT-IIA, and FAS mRNA levels in differentiated
preadipocytes. Preadipocytes from parametrial adipose tissue from
female rats were differentiated for 7 days post-confluence in the
presence of insulin. At day 7 post-confluence, insulin was withdrawn
for 24 h or not, and progesterone treatment (1 µM)
was then performed as above. A representative of two experiments is
shown.
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 |
DISCUSSION |
In the present study, we provide evidence that the expression of
ADD1/SREBP1c mRNA is stimulated by progesterone. We show that high
levels of ADD1/SREBP1c mRNA can be induced upon progesterone stimulation in both the MCF7 cell line and primary cultured
preadipocytes. We show that the effect of progesterone selectively
involves the ADD1/SREBP1c isoform, which can be functionally
distinguished from other cholesterol-regulated SREBP isoforms (for
review, see Ref. 31) and which preferentially targets the expression of lipogenic genes in transgenic mice (17). We also demonstrate that the
progesterone effect on ADD1/SREBP1c is exerted through increased gene
transcription and leads to increased levels of the protein product,
especially the membrane-bound precursor and the mature nuclear
active form of ADD1/SREBP1c. Changes in the levels of active
SREBPs in the nucleus implicate complex post-translational control, such as regulated proteolytic cleavage of the
membrane-bound precursor (31), and also activation through
phosphorylation (32). Our results showing the accumulation of
both the high molecular weight precursor in membranes and the mature
cleaved forms of ADD1/SREBP1 in nuclei upon progesterone treatment
might suggest that proteolytic cleavage is not rate-limiting in the control of ADD1/SREBP1c. However, we cannot exclude the
possibility that progesterone might also activate (directly or
indirectly) ADD1/SREBP1c transcriptional activity. Our present data
agree with a previous report (33) showing that the induction of several lipogenic genes was paralleled by increased levels of the mRNAs encoding SREBP1 and 2 and showing the accumulation of SREBP1 in the nucleus of human prostate cancer cells upon androgen treatment. These observations suggest that the SREBP factors might be a common control point through which sex hormones might signal for metabolic effects. Here we provide direct evidence that nuclear ADD1/SREBP1c protein is the factor through which the stimulatory effect of progesterone on FAS is exerted. This is supported mainly by the fact
that the progesterone effect on FAS mRNA can be abolished by
adenovirus-mediated overexpression of a dominant negative form of ADD1
within the cells, demonstrating that transcriptionally active
ADD1/SREBP1c is required for the progesterone action on the FAS gene.
This suggests that the effect of the hormone is not mediated through
progesterone receptor target DNA sequences, which have not been found
in the 5' regulatory region of the FAS gene. Moreover, in the context
of the regulation of ADD1/SREBP1c expression by insulin, the present
study points out that the expression of ADD1/SREBP1c might be a key
control point to which several hormone signaling pathways might
converge for the regulation of lipogenesis. This is supported by our
data showing that progesterone can act as insulin, to stimulate
ADD1/SREBP1c expression. The effect of these two hormones appear to be
mutually exclusive, at least in the cultured preadipocyte system, which
exhibits both insulin and progesterone sensitivity. This is in
agreement with the fact that both hormones act with similar kinetics on
ADD1/SREBP1c mRNA, through the same mechanism, i.e.
stimulation of ADD1/SREBP1 gene transcription (present data and 19).
The finding of the present study that progesterone might be able in
some situations to replace insulin for the control of ADD1/SREBP1c
expression might have some physiological significance. During late
pregnancy, an insulin-resistant state (34, 35) develops, and maternal
glucose utilization is reduced, hence sparing carbohydrates for the
rapidly growing fetus. However, lipogenesis in the parametrial adipose
tissue remains active (36). In this context, the induction of
ADD1/SREBP1c by progesterone might serve to maintain lipogenesis in
maternal adipose tissue, to preserve energy fat stores required for
lactation, a highly energy-consuming process. On the other hand, it has
been observed that progesterone treatment of diabetic rats was able to
induce lipogenesis in fat (37), further suggesting that in the absence
of insulin, progesterone can serve as an alternative stimulating factor
of adipose tissue lipogenesis. Thus, our present observation that
ADD1/SREBP1c is a progesterone-regulated transcription factor might
provide a mechanism for the understanding of the physiological
regulation of lipogenesis by progesterone.
 |
FOOTNOTES |
*
This work was supported by grants from the European
community program, FAIR 97/3011 and ARC 5858.The costs of publication of this
article were defrayed in part by the
payment of page charges. The article
must therefore be hereby marked
"advertisement" in accordance with 18 U.S.C. Section
1734 solely to indicate this fact.
§
To whom correspondence should be addressed. Tel.: 33 1 42 34 69 22;
Fax: 33 1 40 51 85 86; E-mail: idugail @bhdc.jussieu.fr.
Published, JBC Papers in Press, January 16, 2001, DOI 10.1074/jbc.M008556200
 |
ABBREVIATIONS |
The abbreviations used are:
FAS, fatty acid
synthase;
ADD1, adipocyte determination and differentiation 1;
SREBP, sterol regulatory element-binding protein;
DMEM, Dulbecco's modified
Eagle's medium;
MT-IIA, Metallothionein IIA;
C/EBP, CAAT
enhancer-binding protein.
 |
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