scholarly journals CNBP controls c-Rel dependent IL-12β gene transcription and Th1 immunity

2018 ◽  
Author(s):  
Yongzhi Chen ◽  
Shruti Sharma ◽  
Patricia A. Assis ◽  
Zhaozhao Jiang ◽  
Andrew J. Olive ◽  
...  
Keyword(s):  
Gene Transcription ◽  
Nuclear Translocation ◽  
Binding Activity ◽  
Nucleic Acid Binding ◽  
Th1 Cell ◽  
Inflammatory Gene Expression ◽  
Dna Binding Activity ◽  
Parasite Replication ◽  
Acute Toxoplasmosis ◽  
Selective Impairment

SummaryAn inducible program of inflammatory gene expression is a hallmark of antimicrobial defenses. Herein, we identified Cellular nucleic acid binding protein (Cnbp) as a specific regulator of interleukin-12β gene transcription and Th1 immunity. Cnbp resides in the cytosol of macrophages and translocates to the nucleus in response to a broad range of microbial ligands. Cnbp-deficient macrophages had a selective impairment in their ability to induce IL12β gene transcription. Cnbp interacted with c-Rel, an NFκB/Rel family member that controls IL12β gene transcription. c-Rel nuclear translocation and DNA binding activity were dependent on Cnbp. Furthermore, Cnbp itself bound the IL12β promoter. Lastly, Cnbp-deficient mice were more susceptible to acute toxoplasmosis associated with reduced production of IL12β, as well as a reduced Th1 cell IFNγ response essential to control parasite replication. Collectively, these findings identify Cnbp as a key regulator of c-Rel dependent IL12β gene transcription and Th1 immunity.

scholarly journals CNBP controls IL-12 gene transcription and Th1 immunity

2018 ◽  
Vol 215 (12) ◽  
pp. 3136-3150 ◽  
Author(s):  
Yongzhi Chen ◽  
Shruti Sharma ◽  
Patricia A. Assis ◽  
Zhaozhao Jiang ◽  
Roland Elling ◽  
...  
Keyword(s):  
Gene Expression ◽  
Gene Transcription ◽  
Nuclear Translocation ◽  
Gene Signature ◽  
Binding Activity ◽  
Microbial Pathogens ◽  
Cytokine Gene Expression ◽  
Nucleic Acid Binding ◽  
Th1 Cell ◽  
Dna Binding Activity

An inducible program of inflammatory gene expression is a hallmark of antimicrobial defenses. Recently, cellular nucleic acid–binding protein (CNBP) was identified as a regulator of nuclear factor-kappaB (NF-κB)–dependent proinflammatory cytokine gene expression. Here, we generated mice lacking CNBP and found that CNBP regulates a very restricted gene signature that includes IL-12β. CNBP resides in the cytosol of macrophages and translocates to the nucleus in response to diverse microbial pathogens and pathogen-derived products. Cnbp-deficient macrophages induced canonical NF-κB/Rel signaling normally but were impaired in their ability to control the activation of c-Rel, a key driver of IL-12β gene transcription. The nuclear translocation and DNA-binding activity of c-Rel required CNBP. Lastly, Cnbp-deficient mice were more susceptible to acute toxoplasmosis associated with reduced production of IL-12β, as well as a reduced T helper type 1 (Th1) cell IFN-γ response essential to controlling parasite replication. Collectively, these findings identify CNBP as important regulator of c-Rel–dependent IL-12β gene transcription and Th1 immunity.

scholarly journals 16K prolactin induces NF-kappaB activation in pulmonary fibroblasts

2002 ◽  
Vol 175 (3) ◽  
pp. R13-R18 ◽  
Author(s):  
Y Macotela ◽  
C Mendoza ◽  
AM Corbacho ◽  
G Cosio ◽  
JP Eiserich ◽  
...  
Keyword(s):  
Nitric Oxide ◽  
Nuclear Translocation ◽  
Signal Transduction Pathway ◽  
Binding Activity ◽  
Inos Expression ◽  
Pulmonary Fibroblasts ◽  
Amino Terminal ◽  
Dna Binding Activity ◽  
Nf Kappab ◽  
Oxide Synthase

The amino-terminal 16 kDa fragment of prolactin (16K PRL) promotes the expression of the inducible isoform of nitric oxide synthase (iNOS) accompanied by the production of nitric oxide (NO) by rat pulmonary fibroblasts. The present study was designed to elucidate whether the mechanism by which 16K PRL promotes iNOS expression involves the activation of nuclear factor-kappa B (NF-kappaB), a key transcription factor for iNOS induction. 16K PRL stimulated DNA-binding activity of NF-kappaB in pulmonary fibroblasts as demonstrated by gel shift assays. Likewise, fluorescence immunocytochemistry showed that 16K PRL promotes nuclear translocation of the p65 subunit of NF-kappaB. Finally, treatment with 16K PRL induced the degradation of the NF-kappaB inhibitor kappaB-beta (IkappaB-beta), and such degradation was prevented by blocking IkappaB-beta phosphorylation. Altogether, these results show that 16K PRL activates NF-kappaB nuclear translocation via the phosphorylation and degradation of IkappaB-beta. These findings are consistent with NF-kappaB being part of the signal transduction pathway activated by 16K PRL to induce iNOS expression.

scholarly journals Methylation of STAT6 Modulates STAT6 Phosphorylation, Nuclear Translocation, and DNA-Binding Activity

2004 ◽  
Vol 172 (11) ◽  
pp. 6744-6750 ◽  
Author(s):  
Weiguo Chen ◽  
Michael O. Daines ◽  
Gurjit K. Khurana Hershey
Keyword(s):  
Dna Binding ◽  
Nuclear Translocation ◽  
Binding Activity ◽  
Dna Binding Activity

Silencing of the DEK gene induces apoptosis and senescence in CaSki cervical carcinoma cells via the up-regulation of NF-κB p65

2012 ◽  
Vol 32 (3) ◽  
pp. 323-332 ◽  
Author(s):  
Kuiran Liu ◽  
Tianda Feng ◽  
Jie Liu ◽  
Ming Zhong ◽  
Shulan Zhang
Keyword(s):  
Cell Cycle ◽  
Nuclear Translocation ◽  
Annexin V ◽  
Nuclear Factor Κb ◽  
Binding Activity ◽  
Cell Senescence ◽  
Stable Cell Line ◽  
Reverse Transcription Pcr ◽  
Dna Binding Activity ◽  
Nuclear Extracts

The human DEK proto-oncogene has been found to play an important role in autoimmune disease, viral infection and human carcinogenesis. Although it is transcriptionally up-regulated in cervical cancer, its intracellular function and regulation is still unexplored. In the present study, DEK and IκBα [inhibitor of NF-κB (nuclear factor κB) α] shRNAs (short hairpin RNAs) were constructed and transfected into CaSki cells using Lipofectamine™. The stable cell line CaSki–DEK was obtained after G418 selection. CaSki–IκB cells were observed at 48 h after psiRNA-IκB transfection. The inhibitory efficiency of shRNAs were detected by RT (reverse transcription)–PCR and Western blot analysis. The proliferation activity of cells were measured using an MTT [3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-2H-tetrazolium bromide] assay, cell apoptosis was measured using an Annexin V/PI (propidium iodide) kit, the cell cycle was analysed by flow cytometry and cell senescence was detected using senescence β-galactosidase staining. The intracellular expression of NF-κB p65 protein was studied by cytochemistry. The expression levels of NF-κB p65, p50, c-Rel, IκBα and phospho-IκBα protein were analysed by immunoblotting in whole-cell lysates, cytosolic fractions and nuclear extracts. The protein expression and activity of p38 and JNK (c-Jun N-terminal kinase) were also assayed. In addition, the NF-κB p65 DNA-binding activity was measured by ELISA. Following the silencing of DEK and IκBα, cell proliferation was inhibited, apoptosis was increased, the cell cycle was blocked in the G0/G1-phase with a corresponding decrease in the G2/M-phase, and cell senescence was induced. All of these effects may be related to the up-regulation of NF-κB p65 expression and its nuclear translocation.

scholarly journals Fatty acid signals in Bacillus megaterium are attenuated by cytochromeP-450-mediated hydroxylation

1997 ◽  
Vol 327 (2) ◽  
pp. 363-368 ◽  
Author(s):  
Neil ENGLISH ◽  
N. A. Colin PALMER ◽  
L. William ALWORTH ◽  
Lan KANG ◽  
Valerie HUGHES ◽  
...  
Keyword(s):  
Fatty Acid ◽  
Gene Transcription ◽  
Bacillus Megaterium ◽  
Phytanic Acid ◽  
Binding Activity ◽  
Protein Accumulation ◽  
Peroxisome Proliferators ◽  
Potent Inducer ◽  
Dna Binding Activity

In previous publications [English, Hughes and Wolf (1994) J. Biol. Chem. 269, 26836-26841; English, Hughes and Wolf (1996) Biochem. J. 316, 279-283], we have demonstrated that peroxisome proliferators and non-steroidal anti-inflammatory drugs are inducers of the cytochrome P-450BM-3 gene in Bacillus megaterium ATCC14581. Their mechanism of action involves binding to and subsequent displacement of the transcriptional repressor, Bm3R1, from its operator site, which results in the activation of cytochrome P-450BM-3 gene transcription. We now present evidence that the branched-chain fatty acid, phytanic acid, is a potent inducer of cytochrome P-450BM-3. We have also observed that phytanic acid and peroxisome proliferators are inducers of Bm3R1 protein accumulation and associated DNA-binding activity. In contrast, several barbiturates, although capable of inducing cytochrome P-450BM-3 and Bm3R1 gene transcription, were unable to induce the Bm3R1 protein. We also demonstrate that cytochrome P-450BM-3 readily oxidizes phytanic acid, and provide evidence that, although the ω-1 hydroxy acid derivatives of phytanic acid can associate with Bm3R1, they do so with an affinity two orders of magnitude lower than the unmodified fatty acid. As a consequence, the ability of the hydroxylated product to induce cytochrome P-450BM-3 gene expression in vivo is markedly reduced. These data collectively suggest that metabolism of fatty acids by cytochrome P-450BM-3 leads to an attenuation of their ability to activate the transcription of the BM-3 operon. This work places the action of bacterial fatty acid hydroxylases in an autoregulatory loop where they may be responsible for the inactivation or clearance of the inducing fatty acid signal.

scholarly journals DNA Damage Induces MDMX Nuclear Translocation by p53-Dependent and -Independent Mechanisms

2002 ◽  
Vol 22 (21) ◽  
pp. 7562-7571 ◽  
Author(s):  
Changgong Li ◽  
Lihong Chen ◽  
Jiandong Chen
Keyword(s):  
Dna Damage ◽  
Stress Response ◽  
Embryonic Development ◽  
Nuclear Translocation ◽  
Binding Activity ◽  
Stable Transfection ◽  
Reading Frame ◽  
Dna Binding Activity ◽  
Mdm2 Expression ◽  
Important Regulator

ABSTRACT The MDM2 homolog MDMX is an important regulator of p53 activity during embryonic development. MDMX inactivation in mice results in embryonic lethality in a p53-dependent fashion. The expression level of MDMX is not induced by DNA damage, and its role in stress response is unclear. We show here that ectopically expressed MDMX is mainly localized in the cytoplasm. DNA damage promotes nuclear translocation of MDMX in cells with or without p53. Coexpression of MDM2 or p53 is sufficient to induce MDMX nuclear translocation, suggesting that activation of p53 and induction of MDM2 expression can contribute to this process. Stable transfection of MDMX into U2OS cells does not alter p53 level but results in reduced p53 DNA-binding activity and reduced MDM2 expression. The ability of ARF (alternate reading frame of INK4a) to activate p53 is also significantly inhibited by expression of MDMX. These results suggest that MDMX function may be regulated by DNA damage. Furthermore, MDMX may complement MDM2 in regulating p53 during embryonic development due to its ability to inhibit p53 in the presence of ARF.

scholarly journals α-MSH inhibits induction of C/EBPβ-DNA binding activity and NOS2 gene transcription in macrophages

2000 ◽  
Vol 57 (6) ◽  
pp. 2239-2248 ◽  
Author(s):  
Ashish K. Gupta ◽  
Rebecca A. Diaz ◽  
Sandra Higham ◽  
Bruce C. Kone
Keyword(s):  
Dna Binding ◽  
Gene Transcription ◽  
Binding Activity ◽  
Dna Binding Activity

Abstract 121: GRK5-NT Regulates the Activity of Calcium-Calmodulin-Dependent Transcription Factors

2012 ◽  
Vol 111 (suppl_1) ◽  
Author(s):  
Daniela Sorriento ◽  
Anna Fusco ◽  
Gaetano Santulli ◽  
Bruno Trimarco ◽  
Guido Iaccarino
Keyword(s):  
Transcription Factors ◽  
Cardiac Hypertrophy ◽  
Nuclear Translocation ◽  
Binding Activity ◽  
Activated T Cells ◽  
Calmodulin Binding ◽  
Amino Terminal ◽  
Calcium Calmodulin Dependent ◽  
Dna Binding Activity

We have demonstrated that the amino terminal domain (GRK5-NT), which includes the RH domain and the region of binding to calmodulin, is able to regulate cardiac hypertrophy through inhibition of NFκB. Several studies indicate that mechanical stress and neuro-hormonal activation (Angiotensin, phenylephrine, endothelin) increase the levels of intracellular calcium, therefore activating calcineurin, a serine/threonine phosphatase that dephosphorylates nuclear factor of activated T cells (NFAT). Aim of this study is to evaluate the possibility that GRK5-NT regulates calcium-calmodulin dependent transcription factors. To this aim, we infected cardiomyoblasts in culture with an adenovirus encoding for GRK5-NT (AdGRK5-NT) or treated with a synthetic protein (TAT-RH), which reproduce the only RH domain of GRK5. Hypertrophic responses were induced by chronic stimulation of α 1 adrenergic receptors with phenylephrine (PE 10 -7 M,24h) and the levels of GATA4 and NFAT3 were assessed by western blot. PE induces activation and nuclear translocation of GATA4 and NFAT3 (NFAT:+38±3%;GATA4:+46±3% vs control), the treatment with AdGRK5-NT, but TAT-RH, reduces this effect (NFAT:-68±5%;GATA4:-56±2% vs PE). These data suggest that GRK5-NT using the NT domain regulates the activation of calcium-calmodulin dependent transcription factors through a mechanism of competition for binding to calmodulin. To confirm these data, we evaluated the effect of GRK5-NT in vivo in spontaneously hypertensive and hypertrophic rats (SHR). The overexpression of GRK5-NT in the heart was induced by intracardiac injection of 10 10 pfu/ml of AdGRK5-NT. The treatment inhibits nuclear translocation of NFAT3 and GATA4 (NFAT:-55±1%;GATA4:-34±3% vs PE) and is associated with a reduction in their DNA binding activity, as assessed by EMSA (NFAT:-47±1.5%;GATA4:-33±1% vs EP). In conclusion, our data indicate that GRK5-NT is able to regulate cardiac hypertrophy in two ways: inhibition of NFκB through binding and stabilization of IκB nuclear and inhibition activity of calcium- calmodulin dependent transcription factors, possibly by competing with calmodulin binding.

High glucose-induced NF-κB activation occurs via tyrosine phosphorylation of IκBα in human glomerular endothelial cells: involvement of Syk tyrosine kinase

2008 ◽  
Vol 294 (5) ◽  
pp. F1065-F1075 ◽  
Author(s):  
Won Seok Yang ◽  
Jang Won Seo ◽  
Nam Jeong Han ◽  
Jung Choi ◽  
Ki-Up Lee ◽  
...  
Keyword(s):  
Protein Kinase C ◽  
Protein Kinase ◽  
Tyrosine Phosphorylation ◽  
High Glucose ◽  
Nuclear Translocation ◽  
Binding Activity ◽  
Serine Phosphorylation ◽  
Dna Binding Activity ◽  
Kinase C ◽  
Human Glomerular Endothelial Cells

Activation of nuclear factor-κB (NF-κB) occurs by dissociation from IκB after serine or tyrosine phosphorylation of IκBα, but the way of NF-κB activation by high glucose has not been defined. High glucose is known to activate NF-κB via protein kinase C and reactive oxygen species (ROS). In this study, we investigated how high glucose activates NF-κB for CC chemokine ligand 2 production in cultured human glomerular endothelial cells. High glucose increased nuclear translocation of p65 and also increased NF-κB DNA binding activity. High glucose-induced NF-κB activation occurred without degradation of IκBα. In agreement with this, there was no increase in serine phosphorylation of IκBα, while tyrosine phosphorylation of IκBα was increased by high glucose. High glucose increased the generation of ROS, whereas both α-lipoic acid and N-acetylcysteine scavenged the ROS and decreased high glucose-induced tyrosine phosphorylation of IκBα, nuclear translocation of p65, and NF-κB DNA binding activity. Protein kinase C pseudosubstrate inhibited high glucose-induced ROS production, tyrosine phosphorylation of IκBα, and nuclear translocation of p65. Both BAY 61-3606, a specific inhibitor of Syk protein-tyrosine kinase, and small interfering RNA directed against Syk inhibited high glucose-induced tyrosine phosphorylation of IκBα as well as p65 nuclear translocation. High glucose increased tyrosine phosphorylation of Syk, while it was inhibited by α-lipoic acid and protein kinase C pseudosubstrate. In summary, high glucose-induced NF-κB activation occurred not by serine phosphorylation of IκBα. Our data suggest that ROS-mediated tyrosine phosphorylation of IκBα is the mechanism for high glucose-induced NF-κB activation, and Syk may play a role in tyrosine phosphorylation of IκBα.

Sign in / Sign up

Export Citation Format

Share Document