scholarly journals miR-27b attenuates apoptosis induced by transmissible gastroenteritis virus (TGEV) infection via targeting runt-related transcription factor 1 (RUNX1)

PeerJ ◽  
2016 ◽  
Vol 4 ◽  
pp. e1635 ◽  
Author(s):  
Xiaomin Zhao ◽  
Xiangjun Song ◽  
Xiaoyuan Bai ◽  
Naijiao Fei ◽  
Yong Huang ◽  
...  
Keyword(s):  
Transcription Factor ◽  
Caspase 3 ◽  
Transmissible Gastroenteritis Virus ◽  
Over Expression ◽  
Gastroenteritis Virus ◽  
Related Transcription Factor ◽  
Infected Cells ◽  
Transmissible Gastroenteritis ◽  
Induced Apoptosis ◽  
Transcription Factor 1

Transmissible gastroenteritis virus (TGEV), belonging to the coronaviridae family, is the key cause of the fatal diarrhea of piglets and results in many pathological processes. microRNAs (miRNAs) play a key role in the regulation of virus-induced apoptosis. During the process of apoptosis induced by TGEV infection in PK-15 cells, the miR-27b is notably down-regulated. Thus, we speculate that miR-27b is involved in regulating the process of apoptosis in PK-15 cells. In this study we demonstrated that the over-expression of miR-27b led to the inhibition of TGEV-induced apoptosis, reduction of Bax protein level, and decrease of caspase-3 and −9 activities. Conversely, silencing of miR-27b by miR-27b inhibitors enhanced apoptosis via up-regulating Bax expression and promoting the activities of caspase-3 and −9 in TGEV-infected cells. Subsequently, the runt-related transcription factor 1 (RUNX1) is a candidate target of miR-27b predicted by bioinformatics search. We further identified that the miR-27b directly bound to the 3′ UTR of RUNX1 mRNA and suppressed RUNX1 expression, which indicates RUNX1 is the direct target gene of miR-27b. The over-expression of RUNX1 increased apoptosis and knockdown RUNX1blocked apoptosis of viral-infected cells via regulating Bax expression and the activities of caspase-3 and −9. Our data reveal that miR-27b may repress the mitochondrial pathway of apoptosis by targeting RUNX1, indicating that TGEV may induce apoptosis via down-regulating miR-27b and that miR-27b may act as a target for therapeutic intervention.

scholarly journals Increased Expression of RUNX1 in Liver Correlates with NASH Activity Score in Patients with Non-Alcoholic Steatohepatitis (NASH)

Cells ◽  
2019 ◽  
Vol 8 (10) ◽  
pp. 1277 ◽  
Author(s):  
Kaur ◽  
Rawal ◽  
Siddiqui ◽  
Rohilla ◽  
Sharma ◽  
...  
Keyword(s):  
Transcription Factor ◽  
Activity Score ◽  
Over Expression ◽  
Alcoholic Steatohepatitis ◽  
Related Transcription Factor ◽  
Underlying Mechanisms ◽  
Parenchymal Cells ◽  
Transcription Factor 1

Given the important role of angiogenesis in liver pathology, the current study investigated the role of Runt-related transcription factor 1 (RUNX1), a regulator of developmental angiogenesis, in the pathogenesis of non-alcoholic steatohepatitis (NASH). Quantitative RT-PCRs and a transcription factor analysis of angiogenesis-associated differentially expressed genes in liver tissues of healthy controls, patients with steatosis and NASH, indicated a potential role of RUNX1 in NASH. The gene expression of RUNX1 was correlated with histopathological attributes of patients. The protein expression of RUNX1 in liver was studied by immunohistochemistry. To explore the underlying mechanisms, in vitro studies using RUNX1 siRNA and overexpression plasmids were performed in endothelial cells (ECs). RUNX1 expression was significantly correlated with inflammation, fibrosis and NASH activity score in NASH patients. Its expression was conspicuous in liver non-parenchymal cells. In vitro, factors from steatotic hepatocytes and/or VEGF or TGF- significantly induced the expression of RUNX1 in ECs. RUNX1 regulated the expression of angiogenic and adhesion molecules in ECs, including CCL2, PECAM1 and VCAM1, which was shown by silencing or over-expression of RUNX1. Furthermore, RUNX1 increased the angiogenic activity of ECs. This study reports that steatosis-induced RUNX1 augmented the expression of adhesion and angiogenic molecules and properties in ECs and may be involved in enhancing inflammation and disease severity in NASH.

scholarly journals Overexpression of Myocardin-related transcription factor-A attenuated middle cerebral artery occlusion/reperfusion-induced apoptosis via the Mcl-1/Cyt C/cleaved caspase 3 pathway

2019 ◽  
Vol 12 (06) ◽  
pp. 1950022
Author(s):  
Zhi-Jun Yu ◽  
Jing Du ◽  
Wei Zhang ◽  
Shu-Qi Zhao ◽  
Wei Dong ◽  
...  
Keyword(s):  
Transcription Factor ◽  
Western Blot ◽  
Infarct Size ◽  
Neurological Deficit ◽  
Caspase 3 ◽  
Artery Occlusion ◽  
Related Transcription Factor ◽  
Cyt C ◽  
Induced Apoptosis ◽  
Cerebral Artery Occlusion

To investigate the effect of Myocardin-related transcription factor A (MRTF-A) on apoptosis induced by ischemic/reperfusion (I/R), middle cerebral artery occlusion/reperfusion (MCAO/R) in rats were applied to mimic I/R. The neurological deficit score, cerebral infarct size, cortical neuron apoptosis and cleaved caspase 3 level were evaluated to determine the effect and the level of apoptosis by TTC straining, terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) straining, Western blot and immunofluorescence staining. The myeloid cell leukemia-1 (Mcl-1) expression, release of cytochrome C (Cyt C) and its colocalization with apoptotic protease activating factor-1 (Apaf-1) were analyzed by quantitative real-time PCR (qRT-PCR), Western blot, and immunofluorescence staining. The results showed that MRTF-A overexpression could decrease the neurological deficit score and reduce cerebral infarct size ([Formula: see text] versus Sham). In the MRTF-A-I/R group, TUNEL-positive cells and apoptosis ratio (%) ([Formula: see text]%) were significantly decreased compared to the Neg-I/R group ([Formula: see text]%) at 24[Formula: see text]h reperfusion. Meanwhile, the cleaved caspase 3 expression revealed a similar trend while the expression of Mcl-1 was the opposite. Moreover, MRTF-A overexpression significantly enhanced Mcl-1 fluorescence intensity, which up-regulated the mRNA and protein level ([Formula: see text] or [Formula: see text] versus Neg-I/R). Furthermore, MRTF-A overexpression markedly inhibited the release of Cyt C, and decreased the colocalization with Apaf-1 in the cytoplasm ([Formula: see text] or [Formula: see text] versus Neg-I/R). All the data indicated that MRTF-A overexpression could improve the neurological function against cerebral I/R-induced apoptosis since underlying mechanism might be involved in the Mcl-1/Cyt C/cleaved caspase 3 signaling pathway.

scholarly journals Comparison of a Monoclonal Antibody Capture ELISA (MACELISA) to Indirect ELISA and Virus Neutralization Test for the Serodiagnosis of Transmissible Gastroenteritis Virus

1993 ◽  
Vol 5 (1) ◽  
pp. 21-25 ◽  
Author(s):  
Ignacio Lanza ◽  
Pedro Rubio ◽  
Maria Mufioz ◽  
Pedro Ckmenes
Keyword(s):  
Indirect Elisa ◽  
Neutralization Test ◽  
Virus Neutralization Test ◽  
Virus Neutralization ◽  
Enzyme Linked Immunosorbent Assay ◽  
Transmissible Gastroenteritis Virus ◽  
Gastroenteritis Virus ◽  
Infected Cells ◽  
Transmissible Gastroenteritis ◽  
Sensitivity Specificity

An enzyme-linked immunosorbent assay (ELISA) in which the antigen is captured to the plate by monoclonal antibodies (MACELISA) was developed for the detection of antibodies to transmissible gastroenteritis virus (TGEV). The viral antigen was semipurified from TGEV-infected cells by simple ultracentrifugation. MACELISA results with 258 field sera were compared with those of a standard indirect ELISA and with the virus neutralization test (VNT). Sensitivity, specificity, and kappa values of MACELISA indicated a strong correlation with VNT results, whereas an indirect ELISA was less sensitive and much less specific than VNT. The serologic response of 4 pigs orally inoculated and intraperitoneally boostered with TGEV was compared using the 3 tests. Its sensitivity, specificity, and ability to use unpurified antigen make the MACELISA the advisable first step in TGEV serodiagnosis.

scholarly journals Phosphorylation and subcellular localization of transmissible gastroenteritis virus nucleocapsid protein in infected cells

2005 ◽  
Vol 86 (8) ◽  
pp. 2255-2267 ◽  
Author(s):  
E. Calvo ◽  
D. Escors ◽  
J. A. López ◽  
J. M. González ◽  
A. álvarez ◽  
...  
Keyword(s):  
Mass Spectrometry ◽  
Confocal Microscopy ◽  
Transmissible Gastroenteritis Virus ◽  
Structural Elements ◽  
N Protein ◽  
Gastroenteritis Virus ◽  
Infected Cells ◽  
Transmissible Gastroenteritis ◽  
Secondary Structural Elements ◽  
N Proteins

The nucleocapsid (N) protein is the only phosphorylated structural protein of the coronavirus Transmissible gastroenteritis virus (TGEV). The phosphorylation state and intracellular distribution of TGEV N protein in infected cells were characterized by a combination of techniques including: (i) subcellular fractionation and analysis of tryptic peptides by two-dimensional nano-liquid chromatography, coupled to ion-trap mass spectrometry; (ii) tandem mass-spectrometry analysis of N protein resolved by SDS-PAGE; (iii) Western blotting using two specific antisera for phosphoserine-containing motifs; and (iv) confocal microscopy. A total of four N protein-derived phosphopeptides were detected in mitochondria–Golgi–endoplasmic reticulum–Golgi intermediate compartment (ERGIC)-enriched fractions, including N-protein phosphoserines 9, 156, 254 and 256. Confocal microscopy showed that the N protein found in mitochondria–Golgi–ERGIC fractions localized within the Golgi–ERGIC compartments and not with mitochondria. Phosphorylated N protein was also present in purified virions, containing at least phosphoserines 156 and 256. Coronavirus N proteins showed a conserved pattern of secondary structural elements, including six β-strands and four α-helices. Whilst serine 9 was present in a non-conserved domain, serines 156, 254 and 256 were localized close to highly conserved secondary structural elements within the central domain of coronavirus N proteins. Serine 156 was highly conserved, whereas no clear homologous sites were found for serines 254 and 256 for other coronavirus N proteins.

Sign in / Sign up

Export Citation Format

Share Document