scholarly journals Paralogous Vitamin D Receptors in Teleosts: Transition of Nuclear Receptor Function

Endocrinology ◽  
2008 ◽  
Vol 149 (5) ◽  
pp. 2411-2422 ◽  
Author(s):  
Deanna L. Howarth ◽  
Sheran H. W. Law ◽  
Benjamin Barnes ◽  
Julie M. Hall ◽  
David E. Hinton ◽  
...  
Keyword(s):  
Vitamin D ◽  
Nuclear Receptor ◽  
Functional Divergence ◽  
Functional Characterization ◽  
Binding Domain ◽  
Full Length ◽  
Expression Vectors ◽  
Genome Database ◽  
Vitamin D Receptors ◽  
Medaka Genome

The availability of multiple teleost (bony fish) genomes is providing unprecedented opportunities to understand the diversity and function of gene duplication events using comparative genomics. Here we describe the cloning and functional characterization of two novel vitamin D receptor (VDR) paralogs from the freshwater teleost medaka (Oryzias latipes). VDR sequences were identified through mining of the medaka genome database in which gene organization and structure was determined. Two distinct VDR genes were identified in the medaka genome and mapped to defined loci. Each VDR sequence exhibits unique intronic organization and dissimilar 5′ untranslated regions, suggesting they are not isoforms of the same gene locus. Phylogenetic comparison with additional teleosts and mammalian VDR sequences illustrate that two distinct clusters are formed separating aquatic and terrestrial species. Nested within the teleost cluster are two separate clades for VDRα and VDRβ. The topology of teleost VDR sequences is consistent with the notion of paralogous genes arising from a whole genome duplication event prior to teleost radiation. Functional characterization was conducted through the development of VDR expression vectors including Gal4 chimeras containing the yeast Gal4 DNA binding domain fused to the medaka VDR ligand binding domain and full-length protein. The common VDR ligand 1α,25-dihydroxyvitamin D3 [1α,25(OH)2D3] resulted in significant transactivation activity with both the Gal4 and full-length constructs of medaka (m) VDRβ. Comparatively, transactivation of mVDRα with 1α,25(OH)2D3 was highly attenuated, suggesting a functional divergence between these two nuclear receptor paralogs. We additionally demonstrate through coactivator studies that mVDRα is still functional; however, it exhibits a different sensitivity to 1α,25(OH)2D3, compared with VDRβ. These results suggest that in mVDRα and VDRβ have undergone a functional divergence through a process of sub- and/or neofunctionalization of VDR nuclear receptor gene pairs.

Ligand Binding Domain of Vitamin D Receptors

2006 ◽  
Vol 6 (12) ◽  
pp. 1229-1241 ◽  
Author(s):  
Natacha Rochel ◽  
Dino Moras
Keyword(s):  
Vitamin D ◽  
Ligand Binding ◽  
Binding Domain ◽  
Ligand Binding Domain ◽  
Vitamin D Receptors

Functional characterization of chicken glucocorticoid and mineralocorticoid receptors

2010 ◽  
Vol 298 (5) ◽  
pp. R1257-R1268 ◽  
Author(s):  
Monika Proszkowiec-Weglarz ◽  
Tom E. Porter
Keyword(s):  
Cellular Localization ◽  
Transcriptional Activity ◽  
Functional Characterization ◽  
Nuclear Hormone Receptor ◽  
Avian Species ◽  
Full Length ◽  
Luciferase Assay ◽  
Expression Vectors ◽  
Dependent Manner ◽  
Specific Expression

Glucocorticoid (GR) and mineralocorticoid (MR) receptors are ligand-activated transcription factors that belong to the nuclear hormone receptor superfamily. Little is known about the function of GR and MR in avian species. Recently, the chicken homologue of the GR (cGR) gene was cloned, and its tissue-specific expression was characterized, whereas the full-length sequence of the chicken MR (cMR) gene remains unknown. Therefore, the aims of this project were to clone the full-length cMR and to functionally characterize both chicken receptors. Cos-7 cells were transiently transfected with cGR or cMR expression vectors along with a glucocorticoid response element-luciferase (GRE-Luc) reporter construct. Transfected cells were then treated with increasing doses of corticosterone (CORT) or aldosterone (ALDO) alone and with GR or MR antagonists (ZK98299 and spironolactone, respectively). Transactivation of cGR or cMR was evaluated by luciferase assay. CORT and ALDO induced cGR- and cMR-driven transcriptional activity in a dose-dependent manner. Each receptor responded to both steroids, but cMR transcriptional activity was induced by lower levels of CORT and ALDO than cGR. Coexpression of both chicken corticosteroid receptors in Cos-7 cells had no synergistic or additive effect on CORT- or ALDO-induced transcriptional activity. Corticosteroid-dependent transactivation of cGR and cMR was partially blocked by antagonists. ZK98299 showed high specificity to cGR, while spironolactone had agonist properties toward both receptors. Immunocytochemistry was used to assess the cellular localization of both receptors. Corticosteroids induced translocation of both receptors into the nucleus. The functional properties of cGR and cMR determined in this study will be helpful in defining the physiological roles of GR and MR in avian species.

scholarly journals Polyamines Modulate the Interaction between Nuclear Receptors and Vitamin D Receptor-Interacting Protein 205

2002 ◽  
Vol 16 (7) ◽  
pp. 1502-1510 ◽  
Author(s):  
Yutaka Maeda ◽  
Christophe Rachez ◽  
Leo Hawel ◽  
Craig V. Byus ◽  
Leonard P. Freedman ◽  
...  
Keyword(s):  
Vitamin D ◽  
Ligand Binding ◽  
Nuclear Receptors ◽  
Vitamin D Receptor ◽  
Binding Domain ◽  
Ligand Binding Domain ◽  
Full Length ◽  
Dependent Manner ◽  
Interacting Protein

Abstract Nuclear receptors (NR) activate transcription by interacting with several different coactivator complexes, primarily via LXXLL motifs (NR boxes) of the coactivator that bind a common region in the ligand binding domain of nuclear receptors (activation function-2, AF–2) in a ligand-dependent fashion. However, how nuclear receptors distinguish between different sets of coactivators remains a mystery, as does the mechanism by which orphan receptors such as hepatocyte nuclear factor 4α (HNF4α) activate transcription. In this study, we show that HNF4α interacts with a complex containing vitamin D receptor (VDR)-interacting proteins (DRIPs) in the absence of exogenously added ligand. However, whereas a full-length DRIP205 construct enhanced the activation by HNF4α in vivo, it did not interact well with the HNF4α ligand binding domain in vitro. In investigating this discrepancy, we found that the polyamine spermine significantly enhanced the interaction between HNF4α and full-length DRIP205 in an AF-2, NR-box-dependent manner. Spermine also enhanced the interaction of DRIP205 with the VDR even in the presence of its ligand, but decreased the interaction of both HNF4α and VDR with the p160 coactivator glucocorticoid receptor interacting protein 1 (GR1P1). We also found that GR1P1 and DRIP205 synergistically activated HNF4α-mediated transcription and that a specific inhibitor of polyamine biosynthesis, α-difluoromethylornithine (DFMO), decreased the ability of HNF4α to activate transcription in vivo. These results lead us to propose a model in which polyamines may facilitate the switch between different coactivator complexes binding to NRs.

Assessment of endocrine disruption potential of essential oils of culinary herbs and spices involving glucocorticoid, androgen and vitamin D receptors

2018 ◽  
Vol 9 (4) ◽  
pp. 2136-2144 ◽  
Author(s):  
Iveta Bartoňková ◽  
Zdeněk Dvořák
Keyword(s):  
Vitamin D ◽  
Essential Oils ◽  
Steroid Hormones ◽  
Endocrine Disruption ◽  
Nuclear Receptor ◽  
Vitamin D Receptors ◽  
Culinary Herbs ◽  
Herbs And Spices

Essential oils of culinary herbs and species influence transcriptional activities of nuclear receptor VDR and steroid hormones receptors AR and GR.

scholarly journals The Interaction of TRβ1-N Terminus with Steroid Receptor Coactivator-1 (SRC-1) Serves a Full Transcriptional Activation Function of SRC-1

Endocrinology ◽  
2006 ◽  
Vol 147 (3) ◽  
pp. 1452-1457 ◽  
Author(s):  
Toshiharu Iwasaki ◽  
Akira Takeshita ◽  
Wataru Miyazaki ◽  
William W. Chin ◽  
Noriyuki Koibuchi
Keyword(s):  
Thyroid Hormone ◽  
Nuclear Receptor ◽  
Thyroid Hormone Receptor ◽  
Steroid Receptor ◽  
Activation Function ◽  
Binding Domain ◽  
Expression Vectors ◽  
Response Element ◽  
Steroid Receptor Coactivator ◽  
N Terminus

Steroid receptor coactivator-1 (SRC-1) plays a crucial role in nuclear receptor-mediated transcription including thyroid hormone receptor (TR)-dependent gene expression. Interaction of the TR-ligand binding domain and SRC-1 through LXXLL motifs is required for this action. However, potential interactions between the TRβ1-N terminus (N) and SRC-1 have not been explored and thus are examined in this manuscript. Far-Western studies showed that protein construct containing TRβ1-N + DNA binding domain (DBD) bound to nuclear receptor binding domain (NBD)-1 (amino acid residue, aa 595–780) of SRC-1 without ligand. Mammalian two-hybrid studies showed that NBD-1, as well as SRC-1 (aa 595-1440), bound to TRβ1-N+DBD in the absence of ligand in CV-1 cells. However, NBD-2 (aa 1237–1440) did not bind to this protein. Glutathione-S-transferase pull-down studies showed that TRβ1-N (aa 1–105) bound to the broad region of SRC-1-C terminus. Expression vectors encoding a series of truncations and/or point mutations of TRβ1 were used in transient transfection-based reporter assays in CV-1 cells. N-terminal truncated TRβ1 (ΔN-TRβ1) showed lower activity than that of wild-type in both artificial F2-thyroid hormone response element and native malic enzyme response element. These results suggest that there is the interaction between N terminus of TRβ1 and SRC-1, which may serve a full activation of SRC-1, together with activation function-2 on TRβ1-mediated transcription.

scholarly journals Functional and structural characterization of the insertion region in the ligand binding domain of the vitamin D nuclear receptor

2001 ◽  
Vol 268 (4) ◽  
pp. 971-979 ◽  
Author(s):  
Natachu Rochel ◽  
Guiseppe Tocchini-Valentini ◽  
Pascal F. Egea ◽  
Kari Juntunen ◽  
Jean-Marie Garnier ◽  
...  
Keyword(s):  
Vitamin D ◽  
Ligand Binding ◽  
Nuclear Receptor ◽  
Structural Characterization ◽  
Binding Domain ◽  
Ligand Binding Domain

scholarly journals Molecular Cloning, Functional Characterization, and Evolutionary Analysis of Vitamin D Receptors Isolated from Basal Vertebrates

PLoS ONE ◽  
2015 ◽  
Vol 10 (4) ◽  
pp. e0122853 ◽  
Author(s):  
Erin M. Kollitz ◽  
Guozhu Zhang ◽  
Mary Beth Hawkins ◽  
G. Kerr Whitfield ◽  
David M. Reif ◽  
...  
Keyword(s):  
Vitamin D ◽  
Molecular Cloning ◽  
Functional Characterization ◽  
Evolutionary Analysis ◽  
Vitamin D Receptors

scholarly journals Conformational Changes of RORγ During Response Element Recognition and Coregulator Engagement.

2021 ◽  
Author(s):  
Timothy S Strutzenberg ◽  
Scott J Novick ◽  
Ruben D Garcia-Ordonez ◽  
Christelle Doebelin ◽  
Yuanjun He ◽  
...  
Keyword(s):  
Nuclear Receptor ◽  
Cancer Progression ◽  
Conformational Changes ◽  
Target Gene ◽  
Retinoic Acid Receptor ◽  
Binding Domain ◽  
Full Length ◽  
Structural Proteomics ◽  
Orphan Nuclear Receptor ◽  
Coregulatory Proteins

The retinoic acid receptor-related orphan receptorγ (RORγ) is a ligand-dependent transcription factor of the nuclear receptor super family that underpins metabolic activity, immune function, and cancer progression. Despite being a valuable drug target in health and disease, our understanding of the ligand-dependent activities of RORγ is far from complete. Like most nuclear receptors, RORγ must recruit coregulatory protein to enact the RORγ target gene program. To date, a majority of structural studies have been focused exclusively on the RORγ ligand-binding domain and the ligand-dependent recruitment of small peptide segments of coregulators. Herein, we examine the ligand-dependent assembly of full length RORγ:coregulator complexes on cognate DNA response elements using structural proteomics and small angle x-ray scattering. The results from our studies suggest that RORγ becomes elongated upon DNA recognition, preventing long range interdomain crosstalk. We also determined that the DNA binding domain adopts a sequence-specific conformation, and that coregulatory proteins may be able to sense the ligand- and DNA-bound status of RORγ. We propose a model where ligand-dependent coregulator recruitment may be influenced by the sequence of the DNA to which RORγ is bound. Overall, the efforts described herein will illuminate important aspects of full length RORγ and monomeric orphan nuclear receptor target gene regulation through DNA-dependent conformational changes.

Sign in / Sign up

Export Citation Format

Share Document