Synergistic interactions of silkmoth chorion promoter-binding factors

1991 ◽  
Vol 11 (4) ◽  
pp. 1954-1964
Author(s):  
Y A Skeiky ◽  
K Iatrou
Keyword(s):  
Cell Line ◽  
Transcriptional Activation ◽  
Transcriptional Repression ◽  
Specific Binding ◽  
Ovarian Tissue ◽  
Mobility Shift ◽  
Minimum Requirement ◽  
Synergistic Interactions ◽  
Chorion Genes ◽  
Silkmoth Chorion

Two DNA-binding proteins, BCFI and BCFII, that interact with defined promoter sequences of silkmoth chorion genes of late developmental specificity appear in the nuclei of follicular cells at a time that coincides with the transcriptional activation of the corresponding genes. BCFI prebinding is shown to be indispensable for stable binding of BCFII to its cognate sequence. BCFI and BCFII synergism requires a relatively stringent stereospecific alignment and is a prerequisite for the assembly of higher-order protein-promoter DNA complexes containing additional factors, which are neither gene (stage) nor class (chorion) specific. Binding of BCFI to its site correlates with the induction of DNA structural perturbations that may facilitate assembly of additional factors on the promoter. The BCFI-binding domain contains a core hexanucleotide sequence, AGATAA, which represents the major binding determinant of the erythroid-specific transcription factor GATA-1 of higher vertebrates. This sequence is shown to be necessary and sufficient for binding of BCFI, as it is for a factor that is present in induced K562 human erythroleukemic cells, presumably GATA-1. Comparative analyses of mobility shift patterns obtained with partially proteolyzed preparations of these two unrelated factors were used to confirm that a BCFI-like chorion promoter-binding protein, which is present in the nuclei of an established silkmoth cell line derived from ovarian tissue, is in fact BCFI. The transcriptional repression of endogenous chorion genes in this cell line coupled with the documented absence of factor BCFII suggests that the synergistic interactions between these two factors constitute a minimum requirement for late chorion gene expression.

scholarly journals Synergistic interactions of silkmoth chorion promoter-binding factors.

1991 ◽  
Vol 11 (4) ◽  
pp. 1954-1964 ◽  
Author(s):  
Y A Skeiky ◽  
K Iatrou
Keyword(s):  
Cell Line ◽  
Transcriptional Activation ◽  
Transcriptional Repression ◽  
Specific Binding ◽  
Ovarian Tissue ◽  
Mobility Shift ◽  
Minimum Requirement ◽  
Synergistic Interactions ◽  
Chorion Genes ◽  
Silkmoth Chorion

Two DNA-binding proteins, BCFI and BCFII, that interact with defined promoter sequences of silkmoth chorion genes of late developmental specificity appear in the nuclei of follicular cells at a time that coincides with the transcriptional activation of the corresponding genes. BCFI prebinding is shown to be indispensable for stable binding of BCFII to its cognate sequence. BCFI and BCFII synergism requires a relatively stringent stereospecific alignment and is a prerequisite for the assembly of higher-order protein-promoter DNA complexes containing additional factors, which are neither gene (stage) nor class (chorion) specific. Binding of BCFI to its site correlates with the induction of DNA structural perturbations that may facilitate assembly of additional factors on the promoter. The BCFI-binding domain contains a core hexanucleotide sequence, AGATAA, which represents the major binding determinant of the erythroid-specific transcription factor GATA-1 of higher vertebrates. This sequence is shown to be necessary and sufficient for binding of BCFI, as it is for a factor that is present in induced K562 human erythroleukemic cells, presumably GATA-1. Comparative analyses of mobility shift patterns obtained with partially proteolyzed preparations of these two unrelated factors were used to confirm that a BCFI-like chorion promoter-binding protein, which is present in the nuclei of an established silkmoth cell line derived from ovarian tissue, is in fact BCFI. The transcriptional repression of endogenous chorion genes in this cell line coupled with the documented absence of factor BCFII suggests that the synergistic interactions between these two factors constitute a minimum requirement for late chorion gene expression.

scholarly journals Nuclear Respiratory Factor 1 (NRF-1) Controls the Activity Dependent Transcription of the GABA-A Receptor Beta 1 Subunit Gene in Neurons

2018 ◽  
Author(s):  
Zhuting Li ◽  
Meaghan Cogswell ◽  
Kathryn Hixson ◽  
Amy R. Brooks-Kayal ◽  
Shelley J. Russek
Keyword(s):  
Transcriptional Activation ◽  
Specific Binding ◽  
Core Promoter ◽  
Dominant Negative ◽  
Subunit Gene ◽  
Mobility Shift ◽  
Altered Expression ◽  
Gaba A ◽  
Nuclear Respiratory Factor ◽  
Nuclear Respiratory Factor 1

ABSTRACTWhile the exact role of β1 subunit-containing GABA-A receptors (GABARs) in brain function is not well understood, altered expression of the β1 subunit gene (GABRB1) is associated with neurological and neuropsychiatric disorders. In particular, down-regulation of β1 subunit levels is observed in brains of patients with epilepsy, autism, bipolar disorder, and schizophrenia. A pathophysiological feature of these disease states is imbalance in energy metabolism and mitochondrial dysfunction. The transcription factor, nuclear respiratory factor 1 (NRF-1), has been shown to be a key mediator of genes involved in oxidative phosphorylation and mitochondrial biogenesis. Using a variety of molecular approaches (including mobility shift, promoter/reporter assays, and overexpression of dominant negative NRF-1), we now report that NRF-1 regulates transcription of GABRB1 and that its core promoter contains a conserved canonical NRF-1 element responsible for sequence specific binding and transcriptional activation. Our identification of GABRB1 as a new target for NRF-1 in neurons suggests that genes coding for inhibitory neurotransmission may be coupled to cellular metabolism. This is especially meaningful as binding of NRF-1 to its element is sensitive to the kind of epigenetic changes that occur in multiple disorders associated with altered brain inhibition.

scholarly journals Transcriptional Repression of Peri-Implantation EMX2 Expression in Mammalian Reproduction by HOXA10

2003 ◽  
Vol 23 (1) ◽  
pp. 1-13 ◽  
Author(s):  
Patrick J. Troy ◽  
Gaurang S. Daftary ◽  
Catherine N. Bagot ◽  
Hugh S. Taylor
Keyword(s):  
Transcriptional Activation ◽  
Transcriptional Repression ◽  
Regulatory Region ◽  
Constitutive Expression ◽  
Homeobox Gene ◽  
Urogenital Tract ◽  
Site Directed Mutagenesis ◽  
Northern Analysis ◽  
Mobility Shift ◽  
Regulatory Relationship

ABSTRACT HOXA10 is necessary for mammalian reproduction; however, its transcriptional targets are not completely defined. EMX2, a divergent homeobox gene, is necessary for urogenital tract development. In these studies we identify and characterize the regulation of EMX2 by HOXA10. By using Northern analysis and in situ hybridization, we found that EMX2 is expressed in the adult urogenital tract in an inverse temporal pattern from HOXA10, suggestive of a negative regulatory relationship. Constitutive expression of HOXA10 diminished EMX2 mRNA, whereas blocking HOXA10 through the use of antisense resulted in high EMX2 mRNA expression. Deletional analysis of the EMX2 5′ regulatory region revealed that a 150-bp element mediated transcriptional repression when cotransfected with pcDNA3.1/HOXA10 in transient-transfection assays. Binding of HOXA10 protein to this element was demonstrated by electrophoretic mobility shift assay and further localized to a consensus HOXA10 binding site within this element by DNase I footprinting. Site-directed mutagenesis abolished binding, as well as the negative transcriptional regulation. Transcriptional activation of empty spiracles, the Drosophila ortholog of EMX2, by Abdominal-B (HOXA10 ortholog) has been previously demonstrated. These findings demonstrate conservation of the transcription factor-target gene relationship, although the direction of regulation is reversed with possible evolutionary implications.

scholarly journals Interplay between EGR1 and SP1 is critical for 13-cis retinoic acid-mediated transcriptional repression of angiotensin type 1A receptor

2013 ◽  
Vol 50 (3) ◽  
pp. 361-374 ◽  
Author(s):  
Russell Snyder ◽  
Thomas Thekkumkara
Keyword(s):  
Retinoic Acid ◽  
Protein Expression ◽  
Gene Transcription ◽  
Transcriptional Repression ◽  
De Novo ◽  
Specific Binding ◽  
Consensus Sequence ◽  
Mobility Shift ◽  
Nuclear Extracts ◽  
Angiotensin Type

Recently, we have demonstrated that 13-cis retinoic acid (13cRA) downregulates rat angiotensin type 1A receptor (Agtr1a) gene transcription through a MAP kinase (ERK1/2)-dependent mechanism in rat liver epithelial and aortic smooth muscle cells. However, the exact mechanism remained unknown. In this study, we determined the signaling intermediates activated by ERK1/2 involved in 13cRA-mediated Agtr1a downregulation. Rat Agtr1a chloramphenicol acetyltransferase (CAT) promoter construct containing a sequence -2541 and -1836 bp upstream of the start site demonstrated reduced CAT activity; this region possesses a specificity protein 1 (SP1) consensus sequence (5′-TGGGGCGGGGCGGGG-3′). Mobility shift analysis using untreated nuclear extracts in the presence of mithramycin A suggests that the trans-acting factor binding to this cis-acting element is SP1. 13cRA significantly reduced specific binding without any change in SP1 protein expression. Studies showed that 13cRA treatment maximally phosphorylates ERK1/2 within 5–10 min, which translocates to the nucleus, activating early growth response protein 1 (Egr1) mRNA expression at 20 min followed by de novo protein synthesis, leading to an EGR1/SP1 interaction. siRNA silencing of Egr1 restored Agtr1a mRNA and protein expression in 13cRA-treated cells, and Sp1 silencing results in complete loss of Agtr1a expression. Our study suggests that 13cRA-mediated activation of ERK1/2, through EGR1, is capable of disrupting SP1, the requisite trans-activator for Agtr1a expression, providing a novel paradigm in Agtr1a gene transcription.

scholarly journals Friend of GATA-1–independent transcriptional repression: a novel mode of GATA-1 function

Blood ◽  
2007 ◽  
Vol 109 (12) ◽  
pp. 5230-5233 ◽  
Author(s):  
Kirby D. Johnson ◽  
Meghan E. Boyer ◽  
Jeong-Ah Kang ◽  
Amittha Wickrema ◽  
Alan B. Cantor ◽  
...  
Keyword(s):  
Cell Line ◽  
Transcriptional Activation ◽  
Transcriptional Repression ◽  
Target Genes ◽  
Ex Vivo ◽  
Gata Factor ◽  
Null Cell ◽  
Interacting Protein ◽  
Defective Mutant ◽  
Estrogen Receptor Ligand

Abstract The GATA-1–interacting protein Friend Of GATA-1 (FOG-1) is essential for the proper transcriptional activation and repression of numerous GATA-1 target genes. Although FOG-1–independent activation by GATA-1 has been described, all known examples of GATA-1–mediated repression are FOG-1 dependent. In the GATA-1–null G1E cell line, estrogen receptor ligand binding domain (ER) chimeras of either wild-type GATA-1 or a FOG-1–binding defective mutant of GATA-1 repressed several genes similarly upon activation with β-estradiol. Repression also occurred in a FOG-1–null cell line expressing ER–GATA-1 and during ex vivo erythropoiesis. At the Lyl1 and Rgs18 loci, we found highly restricted occupancy by GATA-1 and GATA-2, indicating that these genes are direct targets of GATA factor regulation. The identification of genes repressed by GATA-1 independent of FOG-1 defines a novel mode of GATA-1–mediated transcriptional regulation.

scholarly journals Dynamic histone acetylation in floral volatile synthesis and emission in petunia flowers

2021 ◽  
Author(s):  
Ryan M Patrick ◽  
Xing-Qi Huang ◽  
Natalia Dudareva ◽  
Ying Li
Keyword(s):  
Transcriptional Activation ◽  
Metabolic Pathways ◽  
Transcriptional Repression ◽  
Metabolic Networks ◽  
Underlying Mechanism ◽  
Expression Of Genes ◽  
Genome Wide ◽  
Chemical Inhibitors ◽  
Volatile Organic ◽  
Floral Volatile

Abstract Biosynthesis of secondary metabolites relies on primary metabolic pathways to provide precursors, energy, and cofactors, thus requiring coordinated regulation of primary and secondary metabolic networks. However, to date, it remains largely unknown how this coordination is achieved. Using Petunia hybrida flowers, which emit high levels of phenylpropanoid/benzenoid volatile organic compounds (VOCs), we uncovered genome-wide dynamic deposition of histone H3 lysine 9 acetylation (H3K9ac) during anthesis as an underlying mechanism to coordinate primary and secondary metabolic networks. The observed epigenome reprogramming is accompanied by transcriptional activation at gene loci involved in primary metabolic pathways that provide precursor phenylalanine, as well as secondary metabolic pathways to produce volatile compounds. We also observed transcriptional repression among genes involved in alternative phenylpropanoid branches that compete for metabolic precursors. We show that GNAT family histone acetyltransferase(s) (HATs) are required for the expression of genes involved in VOC biosynthesis and emission, by using chemical inhibitors of HATs, and by knocking down a specific HAT gene, ELP3, through transient RNAi. Together, our study supports that regulatory mechanisms at chromatin level may play an essential role in activating primary and secondary metabolic pathways to regulate VOC synthesis in petunia flowers.

scholarly journals The plasminogen system and cell surfaces: evidence for plasminogen and urokinase receptors on the same cell type.

1986 ◽  
Vol 103 (6) ◽  
pp. 2411-2420 ◽  
Author(s):  
E F Plow ◽  
D E Freaney ◽  
J Plescia ◽  
L A Miles
Keyword(s):  
Cell Line ◽  
Plasminogen Activator ◽  
Cell Lines ◽  
Specific Binding ◽  
High Capacity ◽  
Fetal Lung ◽  
Fibroblast Cell ◽  
Cell Types ◽  
Cell Surfaces ◽  
Catalytically Active

The capacity of cells to interact with the plasminogen activator, urokinase, and the zymogen, plasminogen, was assessed using the promyeloid leukemic U937 cell line and the diploid fetal lung GM1380 fibroblast cell line. Urokinase bound to both cell lines in a time-dependent, specific, and saturable manner (Kd = 0.8-2.0 nM). An active catalytic site was not required for urokinase binding to the cells, and 55,000-mol-wt urokinase was selectively recognized. Plasminogen also bound to the two cell lines in a specific and saturable manner. This interaction occurred with a Kd of 0.8-0.9 microM and was of very high capacity (1.6-3.1 X 10(7) molecules bound/cell). The interaction of plasminogen with both cell types was partially sensitive to trypsinization of the cells and required an unoccupied high affinity lysine-binding site in the ligand. When plasminogen was added to the GM1380 cells, a line with high intrinsic plasminogen activator activity, the bound ligand was comprised of both plasminogen and plasmin. Urokinase, in catalytically active or inactive form, enhanced plasminogen binding to the two cell lines by 1.4-3.3-fold. Plasmin was the predominant form of the bound ligand when active urokinase was added, and preformed plasmin can also bind directly to the cells. Plasmin on the cell surface was also protected from its primary inhibitor, alpha 2-antiplasmin. These results indicate that the two cell lines possess specific binding sites for plasminogen and urokinase, and a family of widely distributed cellular receptors for these components may be considered. Endogenous or exogenous plasminogen activators can generate plasmin on cell surfaces, and such activation may provide a mechanism for arming cell surfaces with the broad proteolytic activity of this enzyme.

Lipid deprivation increases surfactant phosphatidylcholine synthesis via a sterol-sensitive regulatory element within the CTP:phosphocholine cytidylyltransferase promoter

2002 ◽  
Vol 362 (1) ◽  
pp. 81-88 ◽  
Author(s):  
Rama K. MALLAMPALLI ◽  
Alan J. RYAN ◽  
James L. CARROLL ◽  
Timothy F. OSBORNE ◽  
Christie P. THOMAS
Keyword(s):  
Specific Binding ◽  
Core Promoter ◽  
Regulatory Element ◽  
Simian Virus 40 ◽  
Simian Virus ◽  
Luciferase Reporter ◽  
Standard Diet ◽  
Flanking Sequence ◽  
Mobility Shift ◽  
Ctp:Phosphocholine Cytidylyltransferase

Lipid-deprived mice increase alveolar surfactant disaturated phosphatidylcholine (DSPtdCho) synthesis compared with mice fed a standard diet by increasing expression of CTP:phosphocholine cytidylyltransferase (CCT), the rate-limiting enzyme for DSPtdCho synthesis. We previously observed that lipid deprivation increases mRNA synthesis for CCT [Ryan, McCoy, Mathur, Field and Mallampalli (2000) J. Lipid Res. 41, 1268–1277]. To evaluate regulatory mechanisms for this gene, we cloned the proximal ∼ 1900bp of the 5′ flanking sequence of the murine CCT gene, coupled this to a luciferase reporter, and examined transcriptional regulation in a murine alveolar epithelial type II cell line (MLE-12). The core promoter was localized to a region between −169 and +71bp, which exhibited strong basal activity comparable with the simian virus 40 promoter. The full-length construct, from −1867 to +71, was induced 2–3-fold when cells were cultured in lipoprotein-deficient serum (LPDS), similar to the level of induction of the endogenous CCT gene. By deletional analysis the sterol regulatory element (SRE) was localized within a 240bp region. LPDS activation of the CCT promoter was abolished by mutation of this SRE, and gel mobility-shift assays demonstrated specific binding of recombinant SRE-binding protein to this element within the CCT promoter. These observations indicate that sterol-regulated expression of CCT is mediated by an SRE within its 5′ flanking region.

scholarly journals Transcriptional activation of vesicular monoamine transporter 2 in the pre-B cell line Ea3.123

1999 ◽  
Vol 337 (2) ◽  
pp. 193 ◽  
Author(s):  
Fiona WATSON ◽  
Damian G. DEAVALL ◽  
Janet A. MACRO ◽  
Rachel KIERNAN ◽  
Rod DIMALINE
Keyword(s):  
B Cell ◽  
Cell Line ◽  
Transcriptional Activation ◽  
Vesicular Monoamine Transporter ◽  
Monoamine Transporter ◽  
Vesicular Monoamine Transporter 2

scholarly journals A GM-colony-stimulating factor (CSF) activated ribonuclease system transregulates M-CSF receptor expression in the murine FDC-P1/MAC myeloid cell line.

1992 ◽  
Vol 3 (5) ◽  
pp. 535-544 ◽  
Author(s):  
B C Gliniak ◽  
L S Park ◽  
L R Rohrschneider
Keyword(s):  
Protein Synthesis ◽  
Cell Line ◽  
Transcriptional Activation ◽  
Mrna Degradation ◽  
Receptor Expression ◽  
Metabolic Inhibitors ◽  
Colony Stimulating Factor ◽  
Gm Csf ◽  
Stimulating Factor

The murine myeloid precursor cell line FDC-P1/MAC simultaneously expresses receptors for multi-colony-stimulating factor (CSF), granulocyte-macrophage (GM)-CSF, and macrophage (M)-CSF. Growth of FDC-P1/MAC cells in either multi-CSF or GM-CSF results in the posttranscriptional suppression of M-CSF receptor (c-fms proto-oncogene) expression. We use the term transregulation to describe this control of receptor expression and have further characterized this regulatory process. The removal of FDC-P1/MAC cells from GM-CSF stimulation resulted in the re-expression of c-fms mRNA independent of M-CSF stimulation and new protein synthesis. Switching FDC-P1/MAC cells from growth in M-CSF to GM-CSF caused the selective degradation of c-fms mRNA within 6 h after factor switching. Blocking protein synthesis or gene transcription with metabolic inhibitors effectively prevented GM-CSF stimulated degradation of c-fms mRNA. These results suggest that the transregulation of c-fms transcripts by GM-CSF requires the transcriptional activation of a selective mRNA degradation factor. In vitro analysis, the use of cytoplasmic cell extracts, provided evidence that a ribonuclease is preferentially active in GM-CSF stimulated cells, although the specificity for mRNA degradation in vitro is broader than seen in vivo. Together, these data suggest that GM-CSF can dominantly transregulate the level of c-fms transcript through the transcriptional activation of a ribonuclease degradation system.

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