Functional characterization of WT1 binding sites within the human vitamin D receptor gene promoter

2001 ◽  
Vol 7 (2) ◽  
pp. 187-200 ◽  
Author(s):  
TAE HO LEE ◽  
JERRY PELLETIER
Keyword(s):  
Gene Expression ◽  
Vitamin D ◽  
Transcription Factor ◽  
Vitamin D Receptor ◽  
Binding Sites ◽  
Wilms Tumor ◽  
Cdna Array ◽  
Transcriptional Level ◽  
Vdr Gene ◽  
Downstream Target

The Wilms’ tumor suppressor gene, wt1, encodes a zinc finger transcription factor that can regulate gene expression. It plays an essential role in tumorigenesis, kidney differentiation, and urogenital development. To identify WT1 downstream targets, gene expression profiling was conducted using a cDNA array hybridization approach. We confirm herein that the human vitamin D receptor (VDR), a ligand-activated transcription factor, is a WT1 downstream target. Nuclear run on experiments demonstrated that the effect of WT1 on VDR expression is at the transcriptional level. Transient transfection assays, deletion mutagenesis, electrophoretic mobility shift assays, and chromatin immunoprecipitation assays suggest that, although WT1 is presented with a possibility of three binding sites within the VDR promoter, activation of the human VDR gene appears to occur through a single site. This site differs from a previously identified WT1-responsive site in the murine VDR promoter (Maurer U, Jehan F, Englert C, Hübinger G, Weidmann E, DeLucas HF, and Bergmann L. J Biol Chem 276: 3727–3732, 2001). We also show that the products of a Denys-Drash syndrome allele of wt1 inhibit WT1-mediated transactivation of the human VDR promoter. Our results indicate that the human VDR gene is a downstream target of WT1 and may be regulated differently than its murine counterpart.

scholarly journals Pituitary transcription factor Prop-1 stimulates porcine pituitary glycoprotein hormone α subunit gene expression

2006 ◽  
Vol 37 (2) ◽  
pp. 341-352 ◽  
Author(s):  
Takanobu Sato ◽  
Kousuke Kitahara ◽  
Takao Susa ◽  
Takako Kato ◽  
Yukio Kato
Keyword(s):  
Gene Expression ◽  
Transcription Factor ◽  
Binding Sites ◽  
Gh3 Cells ◽  
Pituitary Hormone ◽  
Transcriptional Level ◽  
Β Subunit ◽  
Glycoprotein Hormone ◽  
Α Subunit

Recently, we have reported that a Prophet of Pit-1 homeodomain factor, Prop-1, is a novel transcription factor for the porcine follicle-stimulating hormone β subunit (FSHβ) gene. This study subsequently aimed to examine the role of Prop-1 in the gene expression of two other porcine gonadotropin subunits, pituitary glycoprotein hormone α subunit (αGSU), and luteinizing hormone β subunit (LHβ). A series of deletion mutants of the porcine αGSU (up to −1059 bp) and LHβ (up to −1277 bp) promoters were constructed in the reporter vector, fused with the secreted alkaline phosphatase gene (pSEAP2-Basic). Transient transfection studies using GH3 cells were carried out to estimate the activation of the porcine αGSU and LHβ promoters by Prop-1, which was found to activate the αGSU promoter of −1059/+12 bp up to 11.7-fold but not the LHβ promoter. Electrophoretic mobility shift assay and DNase I footprinting analysis revealed that Prop-1 binds to six positions, −1038/−1026, −942/−928, −495/−479, −338/−326, −153/−146, and −131/−124 bp, that comprise the A/T cluster. Oligonucleotides of six Prop-1 binding sites were directly connected to the minimum promoter of αGSU, fused in the pSEAP2-Basic vector, followed by transfecting GH3 cells to determine the cis-acting activity. Finally, we concluded that at least five Prop-1 binding sites are the cis-acting elements for αGSU gene expression. The present results revealed a notable feature of the proximal region, where three Prop-1-binding sites are close to and/or overlap the pituitary glycoprotein hormone basal element, GATA-binding element, and junctional regulatory element. To our knowledge, this is the first demonstration of the role of Prop-1 in the regulation of αGSU gene expression. These results, taken together with our previous finding that Prop-1 is a transcription factor for FSHβ gene, confirm that Prop-1 modulates the synthesis of FSH at the transcriptional level. On the other hand, the defects of Prop-1 are known to cause dwarfism and combined pituitary hormone deficiency accompanying hypogonadism. Accordingly, the present observations provide a novel view to understand the hypogonadism caused by Prop-1 defects at the molecular level through the regulatory mechanism of αGSU and FSHβ gene expressions.

scholarly journals Immunological analysis of vitamin D receptor gene expression in Egyptian patients with rheumatoid arthritis: relation to disease activity and functional disability

2020 ◽  
Vol 47 (1) ◽  
Author(s):  
Nevine Mohannad ◽  
Eman Saad Nassar ◽  
Mai Moaaz ◽  
Rehab Elnemr ◽  
Eman Anwar Sultan
Keyword(s):  
Rheumatoid Arthritis ◽  
Gene Expression ◽  
Vitamin D ◽  
Disease Activity ◽  
Vitamin D Receptor ◽  
Functional Disability ◽  
Receptor Gene ◽  
Roc Curve Analysis ◽  
Vdr Gene ◽  
Egyptian Patients

Abstract Background Vitamin D (vit D) deficiency has recently been associated with risk of development of rheumatoid arthritis (RA). The aim of this research was to assess vitamin D receptor (VDR) gene expression in Egyptian patients with RA and its relation with the inflammatory state, disease activity, and functional disability. Results RA patients had significantly lower vit D level and VDR gene expression compared to controls (mean ± 17.0 ± 6.65, 20.73 ± 8.42 ng/ml, p < 0.05 and 3.29 ± 5.47, 14.22 ± 12.60, p < 0.001 respectively). Receiver operating characteristic (ROC) curve analysis for VDR gene expression in RA patients revealed (area under the curve 0.826, cutoff value for low VDR expression 1.05 ng/ml). Patients with low VDR expression had significantly higher ESR, CRP, double positive RF+ anti-CCP+, DAS28, and MHAQ (p < 0.001, p = 0.001, p < 0.05, p < 0.001, p < 0.001) respectively. Conclusion Vitamin D and VDR expression are significantly lower in RA patients than controls. Patients with low VDR gene expression had significantly higher disease activity and disability. This may suggest that apart from low vit D levels, low VDR expression is associated with inflammatory process and it has a potential role in RA pathogenesis and prognosis. Further multicenter studies are needed to confirm these findings.

scholarly journals Vitamin D and Vitamin D Receptor in Scleroderma Subtypes

2020 ◽  
pp. 19-24
Author(s):  
Kocak Ayse
Keyword(s):  
Gene Expression ◽  
Vitamin D ◽  
Vitamin D Receptor ◽  
Mrna Levels ◽  
Diffuse Type ◽  
Gene Expressions ◽  
Vdr Gene ◽  
Vitamin D Levels ◽  
Significant Difference ◽  
Tgf Β1

Vitamin D Receptor (VDR) is a member of the nuclear hormone receptor family. 1,25(OH)2D, a form of metabolically active vitamin D3 form, is the ligand of VDR. When VDR and 1,25(OH)2D are connected, many genes start to molecular interaction reactions that will modulate the transcription. VDR has been shown to be a negative regulator of the transforming growth factor beta-1 / Smad (TGF-β1 / Smad) signalling pathway. TGF-β1 / Smad signalling is important in the pathogenesis of scleroderma (SSc). Vitamin D has pleiotropic effects including immunomodulatory and antifibrotic properties in scleroderma pathogenesis. The aim of this study was to investigate the expression of VDR and the levels of vitamin D in scleroderma subtypes and study the possible correlation between the two parameters. 28 SSc patients and 30 healthy controls were included in the study and they were classified according to the 2013 ACR / EULAR criteria and Rodnan Scores were calculated. 14 were of the limited type and 14 were of the diffuse type of scleroderma. Vitamin D levels were determined in serum. Vitamin D level was measured by chemiluminescence immunometric assay. VDR gene expression was determined by quantitative PCR in isolated RNAs from the blood. Changes in mRNA levels were analysed and beta-actin was used as the housekeeping gene. Also, TGF-β1 gene expressions were determined. VDR gene expressions in diffuse type scleroderma patients were significantly decreased compared to the control. TGF-β1 gene expressions were increased in diffuse type scleroderma. It was found that VDR gene expression in limited type scleroderma patients did not show any significant difference when compared to control. Vitamin D levels and VDR gene expressions showed no correlation in scleroderma subtypes. VDR gene expression decreased in patients with diffuse type scleroderma and showed negative correlation with the Rodnan score and TGF-β1 gene expressions. There was no significant difference between vitamin D and VDR levels.

scholarly journals Altered downstream target gene expression of the placental Vitamin D receptor in human idiopathic fetal growth restriction

Cell Cycle ◽  
2018 ◽  
Vol 17 (2) ◽  
pp. 182-190 ◽  
Author(s):  
Thy P. H. Nguyen ◽  
Hannah E. J. Yong ◽  
Tejasvy Chollangi ◽  
Shaun P. Brennecke ◽  
Susan J. Fisher ◽  
...  
Keyword(s):  
Gene Expression ◽  
Vitamin D ◽  
Vitamin D Receptor ◽  
Fetal Growth ◽  
Fetal Growth Restriction ◽  
Target Gene ◽  
Growth Restriction ◽  
Target Gene Expression ◽  
Downstream Target ◽  
Downstream Target Gene

scholarly journals Methylation Status of Vitamin D Receptor Gene Promoter in Benign and Malignant Adrenal Tumors

2015 ◽  
Vol 2015 ◽  
pp. 1-7 ◽  
Author(s):  
Catia Pilon ◽  
Andrea Rebellato ◽  
Riccardo Urbanet ◽  
Vincenza Guzzardo ◽  
Rocco Cappellesso ◽  
...  
Keyword(s):  
Gene Expression ◽  
Vitamin D ◽  
Vitamin D Receptor ◽  
Methylation Status ◽  
Rare Event ◽  
Gene Promoter ◽  
Benign Tumors ◽  
Cancer Type ◽  
Adrenocortical Tumor ◽  
Vdr Gene

We previously showed a decreased expression of vitamin D receptor (VDR) mRNA/protein in a small group of adrenocortical carcinoma (ACC) tissues, suggesting the loss of a protective role of VDR against malignant cell growth in this cancer type. Downregulation of VDR gene expression may result from epigenetics events, that is, methylation of cytosine nucleotide of CpG islands in VDR gene promoter. We analyzed methylation of CpG sites in the VDR gene promoter in normal adrenals and adrenocortical tumor samples. Methylation of CpG-rich 5′ regions was assessed by bisulfite sequencing PCR using bisulfite-treated DNA from archival microdissected paraffin-embedded adrenocortical tissues. Three normal adrenals and 23 various adrenocortical tumor samples (15 adenomas and 8 carcinomas) were studied. Methylation in the promoter region of VDR gene was found in 3/8 ACCs, while no VDR gene methylation was observed in normal adrenals and adrenocortical adenomas. VDR mRNA and protein levels were lower in ACCs than in benign tumors, and VDR immunostaining was weak or negative in ACCs, including all 3 methylated tissue samples. The association between VDR gene promoter methylation and reduced VDR gene expression is not a rare event in ACC, suggesting that VDR epigenetic inactivation may have a role in adrenocortical carcinogenesis.

scholarly journals Mouse and Human BAC Transgenes Recapitulate Tissue-Specific Expression of the Vitamin D Receptor in Mice and Rescue the VDR-Null Phenotype

Endocrinology ◽  
2014 ◽  
Vol 155 (6) ◽  
pp. 2064-2076 ◽  
Author(s):  
Seong Min Lee ◽  
Kathleen A. Bishop ◽  
Joseph J. Goellner ◽  
Charles A. O'Brien ◽  
J. Wesley Pike
Keyword(s):  
Gene Expression ◽  
Vitamin D ◽  
Vitamin D Receptor ◽  
Null Mouse ◽  
Vdr Gene ◽  
Specific Expression ◽  
Tissue Specific ◽  
Artificial Chromosomes ◽  
Tissue Specific Expression

The biological actions of 1,25-dihydroxyvitamin D3 (1,25(OH)2D3) are mediated by the vitamin D receptor (VDR), which is expressed in numerous target tissues in a cell type-selective manner. Recent studies using genomic analyses and recombineered bacterial artificial chromosomes (BACs) have defined the specific features of mouse and human VDR gene loci in vitro. In the current study, we introduced recombineered mouse and human VDR BACs as transgenes into mice and explored their expression capabilities in vivo. Individual transgenic mouse strains selectively expressed BAC-derived mouse or human VDR proteins in appropriate vitamin D target tissues, thereby recapitulating the tissue-specific expression of endogenous mouse VDR. The mouse VDR transgene was also regulated by 1,25(OH)2D3 and dibutyryl-cAMP. When crossed into a VDR-null mouse background, both transgenes restored wild-type basal as well as 1,25(OH)2D3-inducible gene expression patterns in the appropriate tissues. This maneuver resulted in the complete rescue of the aberrant phenotype noted in the VDR-null mouse, including systemic features associated with altered calcium and phosphorus homeostasis and disrupted production of parathyroid hormone and fibroblast growth factor 23, and abnormalities associated with the skeleton, kidney, parathyroid gland, and the skin. This study suggests that both mouse and human VDR transgenes are capable of recapitulating basal and regulated expression of the VDR in the appropriate mouse tissues and restore 1,25(OH)2D3 function. These results provide a baseline for further dissection of mechanisms integral to mouse and human VDR gene expression and offer the potential to explore the consequence of selective mutations in VDR proteins in vivo.

scholarly journals In Silico Analysis of Regulatory Elements of the Vitamin D Receptor

2020 ◽  
Vol 17 (2) ◽  
pp. 0463
Author(s):  
Shirin Farivar ◽  
Roya Amirinejad ◽  
Bahar Naghavi gargari ◽  
Seyedeh Batool Hassani ◽  
Zeinab Shirvani farsani
Keyword(s):  
Vitamin D ◽  
Transcription Factor ◽  
Vitamin D Receptor ◽  
Protein Interactions ◽  
Molecular Mechanisms ◽  
Cpg Islands ◽  
In Silico Analysis ◽  
Regulatory Elements ◽  
Protein Protein Interactions ◽  
Vdr Gene

Vitamin D receptor (VDR) is a nuclear transcription factor that controls gene expression. Its impaired expression was found to be related to different diseases. VDR also acts as a regulator of different pathways including differentiation, inflammation, calcium and phosphate absorption, etc. but there is no sufficient knowledge about the regulation of the gene itself. Therefore, a better understanding of the genetic and epigenetic factors regulating the VDR may facilitate the improvement of strategies for the prevention and treatment of diseases associated with dysregulation of VDR. In the present investigation, a set of databases and methods were used to identify putative functional elements in the VDR locus. Histone modifications, CpG Islands, epigenetic marks at VDR locus were indicated. In addition, repeated sequences, enhancers, insulators, transcription factor binding sites and targets of the VDR gene, as well as protein-protein interactions with bioinformatics tools, were reported. Some of these genetic elements had overlapped with CpG Islands. These results revealed important new insight into the molecular mechanisms of the VDR gene regulation in human cells and tissues.

Vitamin D Receptor Gene Polymorphisms and the Risk of Metabolic Syndrome (MetS): A Meta-Analysis

2020 ◽  
Vol 20 ◽  
Author(s):  
Hamidreza Totonchi ◽  
Ramazan Rezaei ◽  
Shokoofe Noori ◽  
Negar Azarpira ◽  
Pooneh Mokarram ◽  
...  
Keyword(s):  
Metabolic Syndrome ◽  
Vitamin D ◽  
Vitamin D Receptor ◽  
Protective Factor ◽  
Meta Analysis ◽  
Receptor Gene ◽  
Fixed Effect Model ◽  
Fixed Effect ◽  
Vdr Gene ◽  
Effect Model

Introduction: Several studies have assessed the association between the vitamin D receptor (VDR) polymorphism and risk of metabolic syndrome (MetS). However, the results were inconsistent and inconclusive. Therefore, we conducted a meta-analysis to clarify the exact association between the vitamin D receptor (VDR) polymorphisms and the risk of MetS. Methods: All accessible studies reporting the association between the FokI (rs2228570) or / and TaqI (rs731236) or/and BsmI (rs1544410) or/and ApaI (rs7975232 polymorphisms of the Vitamin D Receptor and susceptibility to MetS published prior to February 2019 were systematically searched in Web of Science, Scopus, and PubMed. After that, Odds ratios (ORs) and their corresponding 95% confidence intervals (CIs) were estimated to evaluate the strength of the association in five genetic models. Results: A total of 9 articles based on four gene variations, and comprising 3348 participants with 1779 metabolic syndrome patients were included. The overall results suggested a significant association between BsmI (rs1544410) polymorphism and MetS susceptibility in recessive model (OR, 0.72, 95% CI, 0.55-0.95, fixed effect model), allelic model (OR, 0.83, 95% CI, 0.72-0.95, fixed effect model), and bb vs BB (OR, 0.65, 95% CI, 0.46-0.93, fixed effect). However, no significant association was identified between TaqI (rs731236) polymorphism, ApaI (rs7975232) polymorphism, and FokI (rs2228570) polymorphism and MetS. Conclusion: This meta-analysis suggested an association between the BsmI (rs1544410) polymorphism and MetS. Indeed, BsmI (rs1544410) acts as a protective factor in the MetS. As a result, the VDR gene could be regarded as a promising pharmacological and physiological target in prevention or treatment of the MetS.

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