scholarly journals EPCR-dependent PAR2 activation by the blood coagulation initiation complex regulates LPS-triggered interferon responses in mice

Blood ◽  
2015 ◽  
Vol 125 (18) ◽  
pp. 2845-2854 ◽  
Author(s):  
Hai Po H. Liang ◽  
Edward J. Kerschen ◽  
Irene Hernandez ◽  
Sreemanti Basu ◽  
Mark Zogg ◽  
...  
Keyword(s):  
Gene Expression ◽  
Blood Coagulation ◽  
Target Genes ◽  
Adaptor Protein ◽  
Messenger Rnas ◽  
Initiation Complex ◽  
Induced Expression ◽  
Regulated Gene Expression

Abstract Infection and inflammation are invariably associated with activation of the blood coagulation mechanism, secondary to the inflammation-induced expression of the coagulation initiator tissue factor (TF) on innate immune cells. By investigating the role of cell-surface receptors for coagulation factors in mouse endotoxemia, we found that the protein C receptor (ProcR; EPCR) was required for the normal in vivo and in vitro induction of lipopolysaccharide (LPS)-regulated gene expression. In cultured bone marrow–derived myeloid cells and in monocytic RAW264.7 cells, the LPS-induced expression of functionally active TF, assembly of the ternary TF-VIIa-Xa initiation complex of blood coagulation, and the EPCR-dependent activation of protease-activated receptor 2 (PAR2) by the ternary TF-VIIa-Xa complex were required for the normal LPS induction of messenger RNAs encoding the TLR3/4 signaling adaptor protein Pellino-1 and the transcription factor interferon regulatory factor 8. In response to in vivo challenge with LPS, mice lacking EPCR or PAR2 failed to fully initiate an interferon-regulated gene expression program that included the Irf8 target genes Lif, Iigp1, Gbp2, Gbp3, and Gbp6. The inflammation-induced expression of TF and crosstalk with EPCR, PAR2, and TLR4 therefore appear necessary for the normal evolution of interferon-regulated host responses.

scholarly journals The TEA Transcription Factor Tec1 Confers Promoter-Specific Gene Regulation by Ste12-Dependent and -Independent Mechanisms

2010 ◽  
Vol 9 (4) ◽  
pp. 514-531 ◽  
Author(s):  
Barbara Heise ◽  
Julia van der Felden ◽  
Sandra Kern ◽  
Mario Malcher ◽  
Stefan Brückner ◽  
...  
Keyword(s):  
Gene Expression ◽  
Transcription Factor ◽  
Transcriptional Activation ◽  
Binding Sites ◽  
Target Genes ◽  
Specific Gene ◽  
Dependent Manner ◽  
A Genome ◽  
Regulated Gene Expression

ABSTRACT In Saccharomyces cerevisiae, the TEA transcription factor Tec1 is known to regulate target genes together with a second transcription factor, Ste12. Tec1-Ste12 complexes can activate transcription through Tec1 binding sites (TCSs), which can be further combined with Ste12 binding sites (PREs) for cooperative DNA binding. However, previous studies have hinted that Tec1 might regulate transcription also without Ste12. Here, we show that in vivo, physiological amounts of Tec1 are sufficient to stimulate TCS-mediated gene expression and transcription of the FLO11 gene in the absence of Ste12. In vitro, Tec1 is able to bind TCS elements with high affinity and specificity without Ste12. Furthermore, Tec1 contains a C-terminal transcriptional activation domain that confers Ste12-independent activation of TCS-regulated gene expression. On a genome-wide scale, we identified 302 Tec1 target genes that constitute two distinct classes. A first class of 254 genes is regulated by Tec1 in a Ste12-dependent manner and is enriched for genes that are bound by Tec1 and Ste12 in vivo. In contrast, a second class of 48 genes can be regulated by Tec1 independently of Ste12 and is enriched for genes that are bound by the stress transcription factors Yap6, Nrg1, Cin5, Skn7, Hsf1, and Msn4. Finally, we find that combinatorial control by Tec1-Ste12 complexes stabilizes Tec1 against degradation. Our study suggests that Tec1 is able to regulate TCS-mediated gene expression by Ste12-dependent and Ste12-independent mechanisms that enable promoter-specific transcriptional control.

scholarly journals Epigenetic regulation of neural cell differentiation plasticity in the adult mammalian brain

2008 ◽  
Vol 105 (46) ◽  
pp. 18012-18017 ◽  
Author(s):  
Jun Kohyama ◽  
Takuro Kojima ◽  
Eriko Takatsuka ◽  
Toru Yamashita ◽  
Jun Namiki ◽  
...  
Keyword(s):  
Gene Expression ◽  
Target Genes ◽  
Epigenetic Modification ◽  
Mammalian Brain ◽  
Cytokine Signaling ◽  
Specific Gene ◽  
Neural Cells ◽  
Specific Gene Expression

Neural stem/progenitor cells (NSCs/NPCs) give rise to neurons, astrocytes, and oligodendrocytes. It has become apparent that intracellular epigenetic modification including DNA methylation, in concert with extracellular cues such as cytokine signaling, is deeply involved in fate specification of NSCs/NPCs by defining cell-type specific gene expression. However, it is still unclear how differentiated neural cells retain their specific attributes by repressing cellular properties characteristic of other lineages. In previous work we have shown that methyl-CpG binding protein transcriptional repressors (MBDs), which are expressed predominantly in neurons in the central nervous system, inhibit astrocyte-specific gene expression by binding to highly methylated regions of their target genes. Here we report that oligodendrocytes, which do not express MBDs, can transdifferentiate into astrocytes both in vitro (cytokine stimulation) and in vivo (ischemic injury) through the activation of the JAK/STAT signaling pathway. These findings suggest that differentiation plasticity in neural cells is regulated by cell-intrinsic epigenetic mechanisms in collaboration with ambient cell-extrinsic cues.

scholarly journals 238.Determination of differentially displayed oxygen-sensitive genes in bovine blastocysts

2004 ◽  
Vol 16 (9) ◽  
pp. 238
Author(s):  
A. J. Harvey ◽  
M. Kirstein ◽  
A. Navarrete-Santos ◽  
K. L. Kind ◽  
B. Fischer ◽  
...  
Keyword(s):  
Gene Expression ◽  
Differential Display ◽  
Target Genes ◽  
Hypoxia Inducible Factor ◽  
Developmental Competence ◽  
Myelogenous Leukemia ◽  
Bovine Embryo ◽  
Nadh Dehydrogenase Subunit 2 ◽  
Regulated Gene Expression

Oxygen-regulated gene expression in the bovine embryo contrasts markedly with that observed in the mouse. Under low (2%) oxygen moderate changes in gene expression are observed in the bovine blastocyst, compared with 3- to 4-fold increases in the mouse. We have determined that these moderate gene expression changes are most likely regulated by Hypoxia-Inducible Factor (HIF)-2 transcription factor activity in the bovine, in the absence of HIF1, although HIF2 target genes are largely unknown. The aim of this study was to screen, by differential display RT-PCR, for putative oxygen-regulated transcripts that might confer developmental competence in blastocysts cultured under varying oxygen atmospheres post compaction. In vitro-produced bovine blastocysts were generated using standard protocols. Compact morulae were randomly allocated to treatments under either 2%, 7% or 20% oxygen for 72 h from Day 5. Blastocyst RNA was isolated using TriReagent and samples were reverse transcribed using Superscript II. cDNA was amplified using 10-mer primers in reactions containing 32Pα-labelled dCTP. Resulting bands were detected by autoradiography, excised, purified and ligated into pGEMT vectors for transformation and sequencing. Seven clones were identified as having high homology with known sequences in GenBank. Real-time PCR was undertaken to confirm oxygen-regulation using Sybr green master mix. Myotrophin mRNA was significantly increased following 2% oxygen culture, compared with 20% cultured blastocysts (P�<�0.01), as was GLUT1 (P�<�0.01). The expression of anaphase-promoting complex showed a significant association with oxygen, being higher in 2% cultured blastocysts (P�<�0.05). Acetyl-coA-acetyltransferase I, chronic myelogenous leukemia tumor antigen (CML66), cyclin I, NADH dehydrogenase subunit 2 and ribonucleotide reductase M1, genes identified using differential display, were not altered by post compaction oxygen concentration. This study has identified potentially HIF2-specific regulated genes, and supports the hypothesis that reduced oxygen concentrations post-compaction may influence bovine embryo development through oxygen-regulated changes in gene expression.

scholarly journals Tissue Distribution of Doxycycline in Animal Models of Tuberculosis

2020 ◽  
Vol 64 (5) ◽  
Author(s):  
Martin Gengenbacher ◽  
Matthew D. Zimmerman ◽  
Jansy P. Sarathy ◽  
Firat Kaya ◽  
Han Wang ◽  
...  
Keyword(s):  
Gene Expression ◽  
Transcriptional Control ◽  
Target Genes ◽  
Calcium Content ◽  
Tissue Penetration ◽  
Lung Lesions ◽  
Content Type ◽  
Clearance Kinetics

ABSTRACT Doxycycline, an FDA-approved tetracycline, is used in tuberculosis in vivo models for the temporal control of mycobacterial gene expression. In these models, animals are infected with recombinant Mycobacterium tuberculosis carrying genes of interest under transcriptional control of the doxycycline-responsive TetR-tetO unit. To minimize fluctuations of plasma levels, doxycycline is usually administered in the diet. However, tissue penetration studies to identify the minimum doxycycline content in food achieving complete repression of TetR-controlled genes in tuberculosis (TB)-infected organs and lesions have not been conducted. Here, we first determined the tetracycline concentrations required to achieve silencing of M. tuberculosis target genes in vitro. Next, we measured doxycycline concentrations in plasma, major organs, and lung lesions in TB-infected mice and rabbits and compared these values to silencing concentrations measured in vitro. We found that 2,000 ppm doxycycline supplemented in mouse and rabbit feed is sufficient to reach target concentrations in TB lesions. In rabbit chow, the calcium content had to be reduced 5-fold to minimize chelation of doxycycline and deliver adequate oral bioavailability. Clearance kinetics from major organs and lung lesions revealed that doxycycline levels fall below concentrations that repress tet promoters within 7 to 14 days after doxycycline is removed from the diet. In summary, we have shown that 2,000 ppm doxycycline supplemented in standard mouse diet and in low-calcium rabbit diet delivers concentrations adequate to achieve full repression of tet promoters in infected tissues of mice and rabbits.

scholarly journals Versatile Retrovirus Vector Systems for Regulated Gene Expression In Vitro and In Vivo

1997 ◽  
Vol 3 (7) ◽  
pp. 466-476 ◽  
Author(s):  
Dirk Lindemann ◽  
Edward Patriquin ◽  
Sandy Feng ◽  
Richard C. Mulligan
Keyword(s):  
Gene Expression ◽  
Vector Systems ◽  
Retrovirus Vector ◽  
Regulated Gene Expression

Abstract 625: Targeting of Heparin Binding EGF-like Growth Factor (HBEGF) Suppresses Hyperlipidemia and Atherosclerosis in LDL Receptor Deficient Mice

2016 ◽  
Vol 36 (suppl_1) ◽  
Author(s):  
Sangderk Lee ◽  
Richard Lee ◽  
Mark Graham ◽  
Lihua Yang ◽  
Seonwook Kim ◽  
...  
Keyword(s):  
Gene Expression ◽  
Growth Factor ◽  
Target Genes ◽  
Regulatory Element ◽  
Atherosclerotic Lesion ◽  
Cell System ◽  
Heparin Binding ◽  
Regulated Gene Expression ◽  
Deficient Mice

Objective: Elevation of apoB-containing lipoproteins is a well-established risk factor for the development of atherosclerosis. Previous reports showed that expression of heparin-binding EGF-like growth factor (HBEGF), a ligand of epidermal growth factor receptor (EGFR), is associated with atherosclerosis development. In this study, we examined in vivo effects of HBEGF targeting on hyperlipidemia-induced atherosclerosis by suppressing HBEGF expression using antisense oligonucleotide (ASO). Methods and Results: Female and male LDLR deficient mice were fed a high fat diet (HFD; 21% fat, 0.2% cholesterol) throughout the study. After 8 weeks of HFD feeding, mice were injected intraperitoneally with either control or HBEGF ASOs weekly for 12 weeks. At termination, we measured circulating lipid concentrations and atherosclerotic lesion size in the aorta. Compared to control ASO group, HBEGF ASO group had a significant reduction of circulating total cholesterol, triglyceride, and apoB-containing lipoprotein concentrations but no change of high-density lipoprotein (HDL) concentration. Importantly, HBEGF ASO injection significantly suppressed atherosclerosis in the aortic arch, thoracic, and abdominal aorta. HBEGF ASO suppressed sterol synthetic gene expression in the liver but elevated lipid contents in the liver. HBEGF gene silencing in a liver cell system induced downregulation of sterol regulatory element binding protein (SREBP) target genes including LDLR and Insig1. Conclusion: Targeting HBEGF using ASOs is an efficient approach to suppress dyslipidemia and hyperlipidemia-induced atherosclerosis. The differential gene expression analysis suggests that HBEGF ASO administration suppresses SREBP-regulated gene expression in the liver leading to downregulation of circulating cholesterol and TG concentrations.

scholarly journals Biosynthetic Enzyme GMP Synthetase Cooperates with Ubiquitin-Specific Protease 7 in Transcriptional Regulation of Ecdysteroid Target Genes

2009 ◽  
Vol 30 (3) ◽  
pp. 736-744 ◽  
Author(s):  
Jan A. van der Knaap ◽  
Elena Kozhevnikova ◽  
Karin Langenberg ◽  
Yuri M. Moshkin ◽  
C. Peter Verrijzer
Keyword(s):  
Gene Expression ◽  
Hormone Receptors ◽  
Target Genes ◽  
Biosynthetic Enzyme ◽  
Histone H2b ◽  
Specific Protease ◽  
Ubiquitin Specific Protease ◽  
Regulated Gene Expression ◽  
Gmp Synthetase

ABSTRACT Drosophila GMP synthetase binds ubiquitin-specific protease 7 (USP7) and is required for its ability to deubiquitylate histone H2B. Previously, we showed that the GMPS/USP7 complex cooperates with the Polycomb silencing system through removal of the active ubiquitin mark from histone H2B (H2Bub). Here, we explored the interplay between GMPS and USP7 further and assessed their role in hormone-regulated gene expression. Genetic analysis established a strong cooperation between GMPS and USP7, which is counteracted by the histone H2B ubiquitin ligase BRE1. Loss of either GMPS or USP7 led to increased levels of histone H2Bub in mutant animals. These in vivo analyses complement our earlier biochemical results, establishing that GMPS/USP7 mediates histone H2B deubiquitylation. We found that GMPS/USP7 binds ecdysone-regulated loci and that mutants display severe misregulation of ecdysone target genes. Ecdysone receptor (EcR) interacts biochemically and genetically with GMPS/USP7. Genetic and gene expression analyses suggested that GMPS/USP7 acts as a transcriptional corepressor. These results revealed the cooperation between a biosynthetic enzyme and a ubiquitin protease in developmental gene control by hormone receptors.

scholarly journals Bile acids inhibit duodenal secretin expression via orphan nuclear receptor small heterodimer partner (SHP)

2009 ◽  
Vol 297 (1) ◽  
pp. G90-G97 ◽  
Author(s):  
Ian P. Y. Lam ◽  
Leo T. O. Lee ◽  
Hueng-Sik Choi ◽  
Gianfranco Alpini ◽  
Billy K. C. Chow
Keyword(s):  
Gene Expression ◽  
Bile Acids ◽  
Nuclear Receptor ◽  
Target Genes ◽  
In Vivo Studies ◽  
Control Group ◽  
Orphan Nuclear Receptor ◽  
Small Heterodimer Partner

Small heterodimer partner (SHP) is an orphan nuclear receptor in which gene expression can be upregulated by bile acids. It regulates its target genes by repressing the transcriptional activities of other nuclear receptors including NeuroD, which has been shown to regulate secretin gene expression. Here, we evaluated the regulation on duodenal secretin gene expression by SHP and selected bile acids, cholic acid (CA) and chenodeoxycholic acid (CDCA). In vitro treatment of CDCA or fexaramine elevated the SHP transcript level and occupancy on secretin promoter. The increase in the SHP level, induced by bile acid treatment or overexpression, reduced secretin gene expression, whereas this gene inhibitory effect was reversed by silencing of endogenous SHP. In in vivo studies, double-immunofluorescence staining demonstrated the coexpression of secretin and SHP in mouse duodenum. Feeding mice with 1% CA-enriched rodent chow resulted in upregulation of SHP and a concomitant decrease in secretin transcript and protein levels in duodenum compared with the control group fed with normal chow. A diet enriched with 5% cholestyramine led to a decrease in SHP level and a corresponding increase in secretin expression. Overall, this study showed that bile acids via SHP inhibit duodenal secretin gene expression. Because secretin is a key hormone that stimulates bile flow in cholangiocytes, this pathway thus provides a novel means to modulate secretin-stimulated choleresis in response to intraduodenal bile acids.

scholarly journals Identification and Verification of the Effect of miR-143 on the Cardioprotective Role of Phosphocreatine in Doxorubicin- Induced Cardiotoxicity by Integrated Bioinformatics Analysis

2021 ◽  
Author(s):  
Chi Zhou ◽  
Zi-Mo Zhou ◽  
Ling Hu ◽  
Ya-Yuan Yang ◽  
Xiang-Wen Meng ◽  
...  
Keyword(s):  
Oxidative Stress ◽  
Gene Expression ◽  
Target Genes ◽  
Bioinformatics Analysis ◽  
Mrna Translation ◽  
Rat Cardiomyocytes ◽  
Adult Rat

Abstract Purpose MicroRNAs (miRNAs) have been reported to play pivotal role in drugs-induced cardiotoxicity act as biomarkes, diagnostic tools and endogenous repressors of gene expression by lowering mRNA stability and interfering with mRNA translation. However, the effect of miRNAs on doxorubicin-induced cardiotoxicity still not clear. In the present study, we identified several key candidate miRNAs involving doxorubicin (DOX)-induced cardiotoxicity in rat myocardial tissues and adult rat cardiomyocytes from the Gene Expression Omnibus (GEO) database via integrated bioinformatics analysis, and the possible effect of miR-143 in the protection of DOX-induced cardiotoxicity by phosphocreatine was subsequently investigated in vivo and in vitro. Methods GSE36239 miRNA expression profiles of DOX-induced cardiotoxicity in rat myocardial tissues and adult rat cardiomyocytes (ARC) were extracted fromGEO datasets. |log2FC| > 1 and P < 0.05 were set as screening criteria, miRNAs expressed in myocardial tissues or ARC were selected as different expression miRNA (DEMs), and subsequently the key miRNAs were obtained from candidate DEMs between myocardial tissues and ARC with Venny 2.1 software. Target genes of miR-143 were predicted with Targetscan and miRBase in the species of homo sapiens, and candidate genes were obtained with Venny 2.1. The gene ontology (GO) and Kyoto Encyclopedia of Genes and Genomes pathway (KEGG) enrichment analyses were carried out. Final, the expression and potential role of miR-143 were verified in DOX-induced cardiotoxicity of rat and cardiomyocytes H9c2. Results A total 24 DEMs were captured , including 15 up-regulated and 9 down-regulated genes in rat myocardial tissues and 42 DEMs were discovered, including 13 up-regulated and 29 down-regulated in ARC. Ultimately, 6 DEMs were determined in rat myocardial tissues and ARC by venny 2.1 software. 46 target genes of miR-143, one of the 6 DEMs, were captured from the predict results of Targetscan and miRBase with venny 2.1. The target genes were notably enriched in biological processes (BP) such as cell proliferation and migration. KEGG pathway analysis showed the target genes were enriched in HIF-1 and PI3K-Akt signaling pathway, which closely related to the oxidative stress and cardiomyocytes apoptosis. Further, western blot and RT-PCR results showed DOX-induced oxidative stress down-regulated the expression of miR-143 and Nrf2, SOD and BCL2, and up-regulated Bax and Cleaved caspase 3, while they could been reversed by the intervention of phosphocreatine (PCr) or N-acetyl-L-cystine (NAC) in DOX-induced cardiotoxicity in vivo and in vitro.Conclusion Our data showed that DOX-induced oxidative stress could decrease the expression of miR-143, promote apoptosis of cardiomyocytes, while PCr or NAC mediated antioxidation could reverse the expression down-regulation of miR-143, alleviated apoptosis in DOX-induced cardiotoxicity. Our findings elucidated the regulatory network involving miR-143 in DOX-induced cardiotoxicity, and might unveiled a potential biomarker and molecular mechanisms, which could be helpful to the diagnosis and treatment of DOX-induced cardiotoxicity.

scholarly journals The m6A demethylase FTO promotes esophageal cancer progression through YTHDF1-dependent posttranscriptional silencing of AKT3

2021 ◽  
Author(s):  
Chunbao Zang ◽  
Fangfang Zhao ◽  
Dabing Huang ◽  
Lingsuo Kong ◽  
Minghua Xie ◽  
...  
Keyword(s):  
Esophageal Cancer ◽  
Cancer Progression ◽  
Target Genes ◽  
Specific Binding ◽  
Messenger Rnas ◽  
Rna Seq ◽  
Functional Studies ◽  
Rna Immunoprecipitation

Abstract Background : N 6 -methyladenosine (m 6 A) is the most abundant modification in eukaryotic messenger RNAs (mRNAs), and plays important roles in many bioprocesses. However, its functions in esophageal cancer remain elusive. Methods : Methylated RNA immunoprecipitation sequencing (MeRIP-seq) and transcriptomic RNA sequencing (RNA-seq) were used to screen the target genes of FTO. Western blot, quantitative real-time PCR (RT-qPCR) and immunohistochemical (IHC) were used to detect FTO expression in cell lines and patient tissues. The biological functions of FTO were investigated in vitro and in vivo . RNA pull-down and RNA immunoprecipitation assays were conducted to explore the specific binding of target genes. Results : We discovered that the RNA demethylase FTO was significantly up-regulated in esophageal cancer patients. Knockdown of FTO drastically reduced esophageal cancer cells (ESCCs) proliferation, migration, invasion, and apoptosis. On the other hand, overexpression of FTO significantly promoted ESCCs growth and invasion. Moreover, we found that the m 6 A methyltransferase METTL14 negatively correlates with FTO function on esophageal cancer progression. By using transcriptome-wide m 6 A-Seq and RNA-Seq assays, we identified AKT3 is the target of FTO, which acts in concert in esophageal cancer tumorigenesis and metastasis. Moreover, loss and gain functional studies confirm that YTHDF1 mediates m 6 A-increased translation of AKT3 mRNA. Conclusion : Our results uncovered an METTL14/FTO/YTHDF1/AKT3 signaling network that regulates the esophageal cancer progression.

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