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1.
Nature ; 534(7605): 124-8, 2016 06 02.
Article in English | MEDLINE | ID: mdl-27251289

ABSTRACT

Liver X receptors (LXRs) are transcriptional regulators of cellular and systemic cholesterol homeostasis. Under conditions of excess cholesterol, LXR activation induces the expression of several genes involved in cholesterol efflux, facilitates cholesterol esterification by promoting fatty acid synthesis, and inhibits cholesterol uptake by the low-density lipoprotein receptor. The fact that sterol content is maintained in a narrow range in most cell types and in the organism as a whole suggests that extensive crosstalk between regulatory pathways must exist. However, the molecular mechanisms that integrate LXRs with other lipid metabolic pathways are incompletely understood. Here we show that ligand activation of LXRs in mouse liver not only promotes cholesterol efflux, but also simultaneously inhibits cholesterol biosynthesis. We further identify the long non-coding RNA LeXis as a mediator of this effect. Hepatic LeXis expression is robustly induced in response to a Western diet (high in fat and cholesterol) or to pharmacological LXR activation. Raising or lowering LeXis levels in the liver affects the expression of genes involved in cholesterol biosynthesis and alters the cholesterol levels in the liver and plasma. LeXis interacts with and affects the DNA interactions of RALY, a heterogeneous ribonucleoprotein that acts as a transcriptional cofactor for cholesterol biosynthetic genes in the mouse liver. These findings outline a regulatory role for a non-coding RNA in lipid metabolism and advance our understanding of the mechanisms that coordinate sterol homeostasis.


Subject(s)
Cholesterol/metabolism , Homeostasis/genetics , Lipid Metabolism/genetics , Orphan Nuclear Receptors/metabolism , RNA, Long Noncoding/genetics , Animals , Cholesterol/biosynthesis , Cholesterol/blood , Diet, Western , Dietary Fats/pharmacology , Gene Expression Regulation , Heterogeneous-Nuclear Ribonucleoprotein Group C/metabolism , Homeostasis/drug effects , Ligands , Lipid Metabolism/drug effects , Liver/drug effects , Liver/metabolism , Liver X Receptors , Male , Mice , Mice, Inbred C57BL , Orphan Nuclear Receptors/agonists , RNA, Long Noncoding/biosynthesis , Signal Transduction , Sterol Regulatory Element Binding Proteins/metabolism
2.
FASEB J ; 25(10): 3416-25, 2011 Oct.
Article in English | MEDLINE | ID: mdl-21676946

ABSTRACT

Human endometrial stromal cells (HESCs) exposed to reactive oxygen species (ROS) mount a hypersumoylation response in a c-Jun N-terminal kinase (JNK)-dependent manner. The mechanism that couples JNK signaling to the small ubiquitin-related modifier (SUMO) pathway and its functional consequences are not understood. We show that ROS-dependent JNK activation converges on the SUMO pathway via PIAS1 (protein inhibitor of activated STAT1). Unexpectedly, PIAS1 knockdown not only prevented ROS-dependent hypersumoylation but also enhanced JNK signaling in HESCs. Conversely, PIAS overexpression increased sumoylation of various substrates, including c-Jun, yet inhibited basal and ROS-dependent JNK activity independently of its SUMO ligase function. Expression profiling demonstrated that PIAS1 knockdown enhances and profoundly modifies the transcriptional response to oxidative stress signals. Using a cutoff of 2-fold change or more, a total of 250 ROS-sensitive genes were identified, 97 of which were not dependent on PIAS1. PIAS1 knockdown abolished the regulation of 43 genes but also sensitized 110 other genes to ROS. Importantly, PIAS1 silencing was obligatory for the induction of several cellular defense genes in response to oxidative stress. In agreement, PIAS1 knockdown attenuated ROS-dependent caspase-3/7 activation and subsequent apoptosis. Thus, PIAS1 determines the level of JNK activity in HESCs, couples ROS signaling to the SUMO pathway, and promotes oxidative cell death.


Subject(s)
Cell Death/physiology , Endometrium/cytology , MAP Kinase Kinase 4/metabolism , Protein Inhibitors of Activated STAT/metabolism , Reactive Oxygen Species/metabolism , Small Ubiquitin-Related Modifier Proteins/metabolism , Cells, Cultured , Female , Gene Expression Regulation/physiology , Humans , MAP Kinase Kinase 4/genetics , Protein Inhibitors of Activated STAT/genetics , Proto-Oncogene Proteins c-jun/genetics , Proto-Oncogene Proteins c-jun/metabolism , Small Ubiquitin-Related Modifier Proteins/genetics , Sumoylation
3.
Biochim Biophys Acta ; 1812(8): 919-28, 2011 Aug.
Article in English | MEDLINE | ID: mdl-21193034

ABSTRACT

Obesity and its associated complications, which can lead to the development of metabolic syndrome, are a worldwide major public health concern especially in developed countries where they have a very high prevalence. RIP140 is a nuclear coregulator with a pivotal role in controlling lipid and glucose metabolism. Genetically manipulated mice devoid of RIP140 are lean with increased oxygen consumption and are resistant to high-fat diet-induced obesity and hepatic steatosis with improved insulin sensitivity. Moreover, white adipocytes with targeted disruption of RIP140 express genes characteristic of brown fat including CIDEA and UCP1 while skeletal muscles show a shift in fibre type composition enriched in more oxidative fibres. Thus, RIP140 is a potential therapeutic target in metabolic disorders. In this article we will review the role of RIP140 in tissues relevant to the appearance and progression of the metabolic syndrome and discuss how the manipulation of RIP140 levels or activity might represent a therapeutic approach to combat obesity and associated metabolic disorders. This article is part of a Special Issue entitled: Translating nuclear receptors from health to disease.


Subject(s)
Adaptor Proteins, Signal Transducing/physiology , Metabolic Syndrome/physiopathology , Nuclear Proteins/physiology , Adipose Tissue/physiopathology , Animals , Humans , Inflammation/physiopathology , Muscles/physiopathology , Nuclear Receptor Interacting Protein 1
4.
FASEB J ; 24(5): 1541-51, 2010 May.
Article in English | MEDLINE | ID: mdl-20026682

ABSTRACT

Survival of the conceptus is dependent on continuous progesterone signaling in the maternal decidua but how this is achieved under conditions of oxidative stress that characterize early pregnancy is unknown. Using primary cultures, we show that modest levels of reactive oxygen species (ROS) increase sumoylation in human endometrial stromal cells (HESCs), leading to enhanced modification and transcriptional inhibition of the progesterone receptor (PR). The ability of ROS to induce a sustained hypersumoylation response, or interfere with PR activity, was lost upon differentiation of HESCs into decidual cells. Hypersumoylation in response to modest levels of ROS requires activation of the JNK pathway. Although ROS-dependent JNK signaling is disabled on decidualization, the cells continue to mount a transcriptional response, albeit distinct from that observed in undifferentiated HESCs. We further show that attenuated JNK signaling in decidual cells is a direct consequence of altered expression of key pathway modulators, including induction of MAP kinase phosphatase 1 (MKP1). Overexpression of MKP1 dampens JNK signaling, prevents hypersumoylation, and maintains PR activity in undifferentiated HESCs exposed to ROS. Thus, JNK silencing uncouples ROS signaling from the SUMO conjugation pathway and maintains progesterone responses and cellular homeostasis in decidual cells under oxidative stress conditions imposed by pregnancy.


Subject(s)
Decidua/metabolism , MAP Kinase Kinase 4/metabolism , Oxidative Stress , Receptors, Progesterone/metabolism , Cell Differentiation , Decidua/cytology , Dual Specificity Phosphatase 1/genetics , Dual Specificity Phosphatase 1/metabolism , Endometrium/metabolism , Female , Gene Expression Regulation , Gene Silencing , Humans , Hydrogen Peroxide/metabolism , MAP Kinase Kinase 4/genetics , Pregnancy , RNA, Small Interfering/genetics , SUMO-1 Protein/metabolism , Signal Transduction , Stromal Cells/metabolism
5.
Endocrinology ; 149(9): 4462-74, 2008 Sep.
Article in English | MEDLINE | ID: mdl-18511503

ABSTRACT

Progesterone is indispensable for differentiation of human endometrial stromal cells (HESCs) into decidual cells, a process that critically controls embryo implantation. We now show an important role for androgen receptor (AR) signaling in this differentiation process. Decreased posttranslational modification of the AR by small ubiquitin-like modifier (SUMO)-1 in decidualizing cells accounted for increased responsiveness to androgen. By combining small interfering RNA technology with genome-wide expression profiling, we found that AR and progesterone receptor (PR) regulate the expression of distinct decidual gene networks. Ingenuity pathway analysis implicated a preponderance of AR-induced genes in cytoskeletal organization and cell motility, whereas analysis of AR-repressed genes suggested involvement in cell cycle regulation. Functionally, AR depletion prevented differentiation-dependent stress fiber formation and promoted motility and proliferation of decidualizing cells. In comparison, PR depletion perturbed the expression of many more genes, underscoring the importance of this nuclear receptor in diverse cellular functions. However, several PR-dependent genes encode for signaling intermediates, and knockdown of PR, but not AR, compromised activation of WNT/beta-catenin, TGFbeta/SMAD, and signal transducer and activator of transcription (STAT) pathways in decidualizing cells. Thus, the nonredundant function of the AR in decidualizing HESCs, centered on cytoskeletal organization and cell cycle regulation, implies an important role for androgens in modulating fetal-maternal interactions. Moreover, we show that PR regulates HESC differentiation, at least in part, by reprogramming growth factor and cytokine signal transduction.


Subject(s)
Decidua/physiology , Endometrium/physiology , Gene Expression Regulation , Gene Regulatory Networks , Receptors, Androgen/physiology , Receptors, Progesterone/physiology , Cells, Cultured , Decidua/metabolism , Endometrium/metabolism , Female , Gene Expression Profiling , Humans , Oligonucleotide Array Sequence Analysis , Protein Processing, Post-Translational , Receptors, Androgen/metabolism , SUMO-1 Protein/metabolism
6.
Am J Hum Genet ; 81(4): 700-12, 2007 Oct.
Article in English | MEDLINE | ID: mdl-17846996

ABSTRACT

The T-box transcription factor TBX22 is essential for normal craniofacial development, as demonstrated by the finding of nonsense, frameshift, splice-site, or missense mutations in patients with X-linked cleft palate (CPX) and ankyloglossia. To better understand the function of TBX22, we studied 10 different naturally occurring missense mutations that are phenotypically equivalent to loss-of-function alleles. Since all missense mutations are located in the DNA-binding T-box domain, we first investigated the preferred recognition sequence for TBX22. Typical of T-box proteins, the resulting sequence is a palindrome based around near-perfect copies of AGGTGTGA. DNA-binding assays indicate that missense mutations at or near predicted contact points with the DNA backbone compromise stable DNA-protein interactions. We show that TBX22 functions as a transcriptional repressor and that TBX22 missense mutations result in impaired repression activity. No effect on nuclear localization of TBX22 was observed. We find that TBX22 is a target for the small ubiquitin-like modifier SUMO-1 and that this modification is required for TBX22 repressor activity. Although the site of SUMO attachment at the lysine at position 63 is upstream of the T-box domain, loss of SUMO-1 modification is consistently found in all pathogenic CPX missense mutations. This implies a general mechanism linking the loss of SUMO conjugation to the loss of TBX22 function. Orofacial clefts are well known for their complex etiology and variable penetrance, involving both genetic and environmental risk factors. The sumoylation process is also subject to and profoundly affected by similar environmental stresses. Thus, we suggest that SUMO modification may represent a common pathway that regulates normal craniofacial development and is involved in the pathogenesis of both Mendelian and idiopathic forms of orofacial clefting.


Subject(s)
Cleft Palate/genetics , Cleft Palate/metabolism , Genes, X-Linked , Genetic Diseases, X-Linked/genetics , Genetic Diseases, X-Linked/metabolism , Mutation, Missense , T-Box Domain Proteins/genetics , Amino Acid Sequence , Animals , Base Sequence , DNA/genetics , DNA/metabolism , Humans , In Vitro Techniques , Male , Mice , Molecular Sequence Data , Mutagenesis, Site-Directed , Phenotype , Recombinant Proteins/genetics , Recombinant Proteins/metabolism , SUMO-1 Protein , Sequence Homology, Amino Acid , Small Ubiquitin-Related Modifier Proteins/genetics , Small Ubiquitin-Related Modifier Proteins/metabolism , T-Box Domain Proteins/metabolism , Transcription, Genetic
7.
Proc Natl Acad Sci U S A ; 103(44): 16272-7, 2006 Oct 31.
Article in English | MEDLINE | ID: mdl-17053081

ABSTRACT

cAMP is required for differentiation of human endometrial stromal cells (HESCs) into decidual cells in response to progesterone, although the underlying mechanism is not well understood. We now demonstrate that cAMP signaling attenuates ligand-dependent sumoylation of the progesterone receptor (PR) in HESCs. In fact, decidualization is associated with global hyposumoylation and redistribution of small ubiquitin-like modifier (SUMO)-1 conjugates into distinct nuclear foci. This altered pattern of global sumoylation was not attributable to impaired maturation of SUMO-1 precursor or altered expression of E1 (SAE1/SEA2) or E2 (Ubc9) enzymes but coincided with profound changes in the expression of E3 ligases and SUMO-specific proteases. Down-regulation of several members of the protein inhibitors of activated STAT (PIAS) family upon decidualization pointed toward a role of these E3 ligases in PR sumoylation. We demonstrate that PIAS1 interacts with the PR and serves as its E3 SUMO ligase upon activation of the receptor. Furthermore, we show that silencing of PIAS1 not only enhances PR-dependent transcription but also induces expression of prolactin, a decidual marker gene, in progestin-treated HESCs without the need of simultaneous activation of the cAMP pathway. Our findings demonstrate how dynamic changes in the SUMO pathway mediated by cAMP signaling determine the endometrial response to progesterone.


Subject(s)
Endometrium/cytology , Endometrium/metabolism , Progesterone/metabolism , SUMO-1 Protein/metabolism , Signal Transduction , Stromal Cells/cytology , Stromal Cells/metabolism , Cell Differentiation , Cells, Cultured , Cyclic AMP/metabolism , Female , Humans , Ligases/metabolism , Protein Binding , Protein C Inhibitor/genetics , Protein C Inhibitor/metabolism , Receptors, Progesterone/genetics , Receptors, Progesterone/metabolism , SUMO-1 Protein/genetics , Transcription, Genetic/genetics
8.
Mol Endocrinol ; 20(1): 35-44, 2006 Jan.
Article in English | MEDLINE | ID: mdl-16123151

ABSTRACT

Menstruation, or cyclic shedding of nonpregnant endometrial tissue with associated bleeding, occurs only in humans and a few other species. This breakdown of the endometrium in response to falling ovarian progesterone levels is a complex process, characterized by local leukocyte infiltration, expression and activation of matrix metalloproteinases, and apoptosis. Spontaneous decidualization (differentiation) of the stromal compartment precedes the cyclic shedding of the endometrium in various menstruating species but the mechanisms that link these processes are not understood. In this study, we identified FOXO1 as a key transcription factor responsible for mediating apoptosis of decidualized human endometrial stromal cells (HESCs) in response to progesterone withdrawal. We demonstrate that medroxyprogesterone acetate (MPA, a synthetic progestin) enhances the expression of FOXO1 in differentiating HESCs while simultaneously inducing cytoplasmic retention and inactivation of FOXO1. Withdrawal of MPA from decidualized HESCs results in rapid nuclear accumulation of FOXO1, increased BIM expression, a proapoptotic FOXO1 target gene, and cell death. Conversely, silencing of FOXO1 expression completely abolishes cell death induced by MPA withdrawal. In summary, the observation that differentiating HESCs become dependent on progesterone signaling for survival through induction and reversible inactivation of FOXO1 suggests a novel mechanism that links decidualization of the endometrium to menstruation.


Subject(s)
Endometrium/cytology , Forkhead Transcription Factors/metabolism , Progestins/physiology , Active Transport, Cell Nucleus , Apoptosis Regulatory Proteins/metabolism , Bcl-2-Like Protein 11 , Cell Differentiation , Cells, Cultured , Cyclic AMP/metabolism , Cytoplasm/metabolism , Decidua/cytology , Decidua/metabolism , Endometrium/metabolism , Female , Forkhead Box Protein O1 , Forkhead Transcription Factors/genetics , Gene Expression Regulation , Humans , Medroxyprogesterone Acetate/pharmacology , Membrane Proteins/metabolism , Progesterone/pharmacology , Proto-Oncogene Proteins/metabolism , RNA, Small Interfering/genetics , Stromal Cells/cytology , Stromal Cells/metabolism
9.
Mol Cell Biol ; 24(22): 10058-71, 2004 Nov.
Article in English | MEDLINE | ID: mdl-15509806

ABSTRACT

Cell cycle arrest by FoxO transcription factors involves transcriptional repression of cyclin D, although the exact mechanism remains unclear. In this study, we used the BCR-ABL-expressing cell line BV173 as a model system to investigate the mechanisms whereby FoxO3a regulates cyclin D2 expression. Inhibition of BCR-ABL by STI571 results in down-regulation of cyclin D2 expression, activation of FoxO3a activity, and up-regulation of BCL6 expression. Using reporter gene assays, we demonstrate that STI571, FoxO3a, and BCL6 can repress cyclin D2 transcription through a STAT5/BCL6 site located within the cyclin D2 promoter. We propose that BCR-ABL inhibition leads to FoxO3a activation, which in turn induces the expression of BCL6, culminating in the repression of cyclin D2 transcription through this STAT5/BCL6 site. This process was verified by mobility shift and chromatin immunoprecipitation analyses. We find that conditional activation of FoxO3a leads to accumulation of BCL6 and down-regulation of cyclin D2 at protein and mRNA levels. Furthermore, silencing of FoxO3a and BCL6 in BCR-ABL-expressing cells abolishes STI571-mediated effects on cyclin D2. This report establishes the signaling events whereby BCR-ABL signals are relayed to cyclin D2 to mediate cell cycle progression and defines a potential mechanism by which FoxO proteins regulate cyclin D2 expression.


Subject(s)
Cyclins/genetics , DNA-Binding Proteins/metabolism , Fusion Proteins, bcr-abl/metabolism , Milk Proteins/metabolism , Trans-Activators/metabolism , Transcription Factors/metabolism , Base Sequence , Benzamides , Binding Sites/genetics , Cell Line , Cyclin D2 , Cyclins/metabolism , DNA, Complementary/genetics , DNA-Binding Proteins/genetics , Forkhead Box Protein O1 , Forkhead Transcription Factors , Fusion Proteins, bcr-abl/antagonists & inhibitors , Fusion Proteins, bcr-abl/genetics , Humans , Imatinib Mesylate , Milk Proteins/genetics , Nuclear Proteins/metabolism , Phosphorylation , Piperazines/pharmacology , Promoter Regions, Genetic , Protein Binding , Protein Kinase Inhibitors/pharmacology , Proto-Oncogene Proteins c-bcl-6 , Pyrimidines/pharmacology , RNA, Messenger/genetics , RNA, Messenger/metabolism , RNA, Small Interfering/genetics , STAT5 Transcription Factor , Signal Transduction , Trans-Activators/genetics , Transcription Factors/genetics , Transcription, Genetic/drug effects
10.
Mol Endocrinol ; 18(8): 1988-99, 2004 Aug.
Article in English | MEDLINE | ID: mdl-15155784

ABSTRACT

All cardinal events during the reproductive cycle, including ovulation, implantation, and menstruation, are characterized by a profound tissue remodeling and an associated local inflammatory response. The ovarian hormone progesterone is a key modulator of inflammatory signals in reproductive tissues, but the underlying mechanisms are not well understood. In this study, we report that differentiating human endometrial stromal cells (ESCs) acquire resistance to interferon-gamma (IFNgamma)-dependent signal transducers and activators of transcription (STAT) 1 signaling, although phosphorylation, nuclear translocation, and binding of STAT1 to DNA, are unaffected. These observations prompted an investigation into the role of nuclear repressors of STAT1 signaling. We demonstrate that protein inhibitor of activated STAT-y is complexed to the progesterone receptor (PR) in human ESCs and that its ability to repress STAT1 signaling is dependent upon activation of PR in response to hormone binding. Conversely, IFNgamma and protein inhibitor of activated STAT-y synergistically inhibited PR-dependent transcription, demonstrating that the progesterone and IFNgamma signaling pathways engage in reciprocal transcriptional antagonism in human endometrium.


Subject(s)
Endometrium/drug effects , Interferon-gamma/pharmacology , Intracellular Signaling Peptides and Proteins/metabolism , Progesterone/pharmacology , Signal Transduction/drug effects , Cell Differentiation , Cells, Cultured , DNA/metabolism , DNA-Binding Proteins/metabolism , Endometrium/cytology , Endometrium/metabolism , Female , Gene Expression Regulation/drug effects , Histone Deacetylase Inhibitors , Histone Deacetylases/metabolism , Humans , Hydroxamic Acids/pharmacology , Phosphorylation/drug effects , Poly-ADP-Ribose Binding Proteins , Protein Inhibitors of Activated STAT , Receptors, Progesterone/antagonists & inhibitors , Receptors, Progesterone/metabolism , STAT1 Transcription Factor , Trans-Activators/metabolism , Transcription, Genetic/genetics
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