scholarly journals Development of BacMam Induced Hepatitis E Virus Replication Model in Hepatoma Cells to Study the Polyprotein Processing

2020 ◽  
Vol 11 ◽  
Author(s):  
Manjeet Kumar ◽  
Preeti Hooda ◽  
Madhu Khanna ◽  
Utkarsh Patel ◽  
Deepak Sehgal
Keyword(s):  
Virus Replication ◽  
Hepatitis E Virus ◽  
Hepatitis E ◽  
Hepatoma Cells ◽  
Polyprotein Processing ◽  
Replication Model

scholarly journals Activities of Thrombin and Factor Xa Are Essential for Replication of Hepatitis E Virus and Are Possibly Implicated in ORF1 Polyprotein Processing

2018 ◽  
Vol 92 (6) ◽  
Author(s):  
Gayatri D. Kanade ◽  
Kunal D. Pingale ◽  
Yogesh A. Karpe
Keyword(s):  
Cell Culture ◽  
Cell Line ◽  
Liver Cell ◽  
Serine Proteases ◽  
Hepatitis E Virus ◽  
Factor Xa ◽  
Hepatitis E ◽  
Cleavage Sites ◽  
Polyprotein Processing ◽  
Liver Cell Line

ABSTRACTHepatitis E virus (HEV) is a clinically important positive-sense RNA virus. The ORF1 of HEV encodes a nonstructural polyprotein of 1,693 amino acids. It is not clear whether the ORF1 polyprotein (pORF1) is processed into distinct enzymatic domains. Many researchers have attempted to understand the mechanisms of pORF1 processing. However, these studies gave various results and could never convincingly establish the mechanism of pORF1 processing. In this study, we demonstrated the possible role of thrombin and factor Xa in pORF1 processing. We observed that the HEV pORF1 polyprotein bears conserved cleavage sites of thrombin and factor Xa. Using a reverse genetics approach, we demonstrated that an HEV replicon having mutations in the cleavage sites of either thrombin or factor Xa could not replicate efficiently in cell culture. Further, we demonstratedin vitroprocessing when we incubated recombinant pORF1 fragments with thrombin, and we observed the processing of pORF1 polyprotein. The treatment of a liver cell line with a serine protease inhibitor as well as small interfering RNA (siRNA) knockdown of thrombin and factor Xa resulted in significant reduction in the replication of HEV. Thrombin and factor Xa have been well studied for their roles in blood clotting. Both of these proteins are believed to be present in the active form in the blood plasma. Interestingly, in this report, we demonstrated the presence of biologically active thrombin and factor Xa in a liver cell line. The results suggest that factor Xa and thrombin are essential for the replication of HEV and may be involved in pORF1 polyprotein processing of HEV.IMPORTANCEHepatitis E virus (HEV) causes a liver disorder called hepatitis in humans, which is mostly an acute and self-limiting infection in adults. A high mortality rate of about 30% is observed in HEV-infected pregnant women in developing countries. There is no convincing opinion about HEV ORF1 polyprotein processing owing to the variability of study results obtained so far. HEV pORF1 has cleavage sites for two host cellular serine proteases, thrombin and factor Xa, that are conserved among HEV genotypes. For the first time, this study demonstrated that thrombin and factor Xa cleavage sites on HEV pORF1 are obligatory for HEV replication. Intracellular biochemical activities of the said serine proteases are also essential for efficient HEV replication in cell culture and must be involved in pORF1 processing. This study sheds light on the presence and roles of clotting factors with respect to virus replication in the cells.

scholarly journals Pan-Genotype Hepatitis E Virus Replication in Stem Cell–Derived Hepatocellular Systems

2018 ◽  
Vol 154 (3) ◽  
pp. 663-674.e7 ◽  
Author(s):  
Xianfang Wu ◽  
Viet Loan Dao Thi ◽  
Peng Liu ◽  
Constantin N. Takacs ◽  
Kuanhui Xiang ◽  
...  
Keyword(s):  
Stem Cell ◽  
Virus Replication ◽  
Hepatitis E Virus ◽  
Hepatitis E

scholarly journals Inhibition of hepatitis E virus replication by peptide-conjugated morpholino oligomers

2015 ◽  
Vol 120 ◽  
pp. 134-139 ◽  
Author(s):  
Yuchen Nan ◽  
Zexu Ma ◽  
Harilakshmi Kannan ◽  
David A. Stein ◽  
Patrick I. Iversen ◽  
...  
Keyword(s):  
Virus Replication ◽  
Hepatitis E Virus ◽  
Hepatitis E

scholarly journals Evidence of MAPK–JNK1/2 activation by hepatitis E virus ORF3 protein in cultured hepatoma cells

2014 ◽  
Vol 67 (3) ◽  
pp. 545-550 ◽  
Author(s):  
Mohammad Khalid Parvez ◽  
Mohammed Salem Al-Dosari
Keyword(s):  
Hepatitis E Virus ◽  
Hepatitis E ◽  
Hepatoma Cells ◽  
Orf3 Protein

RIG-I Inhibits Hepatitis E Virus Replication by Simultaneously Activating JAK-STAT and NFKB Pathways

2016 ◽  
Vol 64 (2) ◽  
pp. S518
Author(s):  
L. Xu ◽  
W. Wang ◽  
X. Zhou ◽  
Y. Wang ◽  
Y. Yin ◽  
...  
Keyword(s):  
Virus Replication ◽  
Hepatitis E Virus ◽  
Hepatitis E

scholarly journals Hepatitis E virus replication involves alternating negative- and positive-sense RNA synthesis

2010 ◽  
Vol 92 (3) ◽  
pp. 572-581 ◽  
Author(s):  
S. P. K. Varma ◽  
A. Kumar ◽  
N. Kapur ◽  
H. Durgapal ◽  
S. K. Acharya ◽  
...  
Keyword(s):  
Virus Replication ◽  
Hepatitis E Virus ◽  
Rna Synthesis ◽  
Hepatitis E ◽  
Positive Sense

2’-C-methylcytidine inhibits hepatitis E virus replication but antagonizing ribavirin

2017 ◽  
Vol 66 (1) ◽  
pp. S471-S472
Author(s):  
C. Qu ◽  
W. Wang ◽  
L. Xu ◽  
Y. Yin ◽  
Q. Pan ◽  
...  
Keyword(s):  
Virus Replication ◽  
Hepatitis E Virus ◽  
Hepatitis E

scholarly journals Full-length MAVS Inhibits Hepatitis E Virus Replication Dependent of JAK Signaling

2021 ◽  
Author(s):  
Changbo Qu ◽  
Yang Li ◽  
Yunlong Li ◽  
Yihang Pan
Keyword(s):  
Acute Hepatitis ◽  
Virus Replication ◽  
Hepatitis E Virus ◽  
Hepatitis E ◽  
Antiviral Signaling ◽  
Interferon Stimulated Genes ◽  
Mitochondrial Antiviral Signaling ◽  
Signaling Activation ◽  
Transcriptional Induction ◽  
Mitochondrial Antiviral Signaling Protein

Abstract Hepatitis E virus (HEV) infection is the leading cause of acute hepatitis worldwide. Mitochondrial antiviral signaling protein (MAVS)-mediated interferon (IFN) response plays a pivotal role in the hepatic antiviral immunity. However, little is known about the effects of overexpression of MAVS on HEV infection. Here, we studied the effects of FL-MAVS on HEV. We found that overexpression of FL-MAVS profoundly inhibited HEV replication. The overexpression of FL-MAVS is accompanied by the secretion of functional IFNs and transcriptional induction of interferon stimulated genes (ISGs). Furthermore, we showed that the anti-HEV effect of FL-MAVS is largely dependent of the JAK signaling activation.

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