Hepatitis C Virus Non-structural Proteins in the Probable Membranous Compartment Function in Viral Genome Replication

Hepatitis C virus (HCV) non-structural protein 5A (NS5A) plays an essential role in viral genome replication. A series of transposon-mediated insertion mutants and deletion mutants of NS5A was used to examine the colony-forming ability of HCV subgenomic replicons encoding the mutant proteins. The results reveal that two regions of NS5A can tolerate insertions: one spanning residues 240–314, which contain the interferon sensitivity-determining region (ISDR), and the other spanning residues 349–417 at the carboxy terminus. The majority of these sites also tolerated insertion of enhanced green fluorescent protein. Furthermore, replicons encoding NS5A with deletions in ISDR or in the carboxy-terminal regions were replication-competent, indicating that these regions of NS5A are not necessary for replication. Taken together, the results suggest that the central region spanning the ISDR and the carboxy-terminal region of the molecule are dispensable for the functions of NS5A in viral genome replication.

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Hepatitis C Virus Core-Derived Peptides Inhibit Genotype 1b Viral Genome Replication via Interaction with DDX3X

PLoS ONE ◽

10.1371/journal.pone.0012826 ◽

2010 ◽

Vol 5 (9) ◽

pp. e12826 ◽

Cited By ~ 14

Author(s):

Chaomin Sun ◽

Cara T. Pager ◽

Guangxiang Luo ◽

Peter Sarnow ◽

Jamie H. D. Cate

Keyword(s):

Hepatitis C Virus ◽

Hepatitis C ◽

Viral Genome ◽

Genome Replication ◽

Virus Core ◽

Genotype 1B ◽

Viral Genome Replication

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Involvement of Creatine Kinase B in Hepatitis C Virus Genome Replication through Interaction with the Viral NS4A Protein

Journal of Virology ◽

10.1128/jvi.02179-08 ◽

2009 ◽

Vol 83 (10) ◽

pp. 5137-5147 ◽

Cited By ~ 30

Author(s):

Hiromichi Hara ◽

Hideki Aizaki ◽

Mami Matsuda ◽

Fumiko Shinkai-Ouchi ◽

Yasushi Inoue ◽

...

Keyword(s):

Creatine Kinase ◽

Hepatitis C Virus ◽

Hepatitis C ◽

Viral Genome ◽

Proteome Analysis ◽

Virus Genome ◽

Genome Replication ◽

Creatine Kinase B ◽

Efficient Replication

ABSTRACT Persistent infection with hepatitis C virus (HCV) is a major cause of chronic liver diseases. The aim of this study was to identify host cell factor(s) participating in the HCV replication complex (RC) and to clarify the regulatory mechanisms of viral genome replication dependent on the host-derived factor(s) identified. By comparative proteome analysis of RC-rich membrane fractions and subsequent gene silencing mediated by RNA interference, we identified several candidates for RC components involved in HCV replication. We found that one of these candidates, creatine kinase B (CKB), a key ATP-generating enzyme that regulates ATP in subcellular compartments of nonmuscle cells, is important for efficient replication of the HCV genome and propagation of infectious virus. CKB interacts with HCV NS4A protein and forms a complex with NS3-4A, which possesses multiple enzyme activities. CKB upregulates both NS3-4A-mediated unwinding of RNA and DNA in vitro and replicase activity in permeabilized HCV replicating cells. Our results support a model in which recruitment of CKB to the HCV RC compartment, which has high and fluctuating energy demands, through its interaction with NS4A is important for efficient replication of the viral genome. The CKB-NS4A association is a potential target for the development of a new type of antiviral therapeutic strategy.

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Suppression of Hepatitis C Viral Genome Replication with RNA-Cleaving Deoxyribozyme

From Nucleic Acids Sequences to Molecular Medicine ◽

10.1007/978-3-642-27426-8_17 ◽

2012 ◽

pp. 429-452 ◽

Cited By ~ 2

Author(s):

Dal-Hee Min ◽

Dong-Eun Kim

Keyword(s):

Hepatitis C ◽

Viral Genome ◽

Genome Replication ◽

Viral Genome Replication

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Suppression of Hepatitis C Virus Genome Replication and Particle Production by a Novel Diacylglycerol Acyltransferases Inhibitor

Molecules ◽

10.3390/molecules23082083 ◽

2018 ◽

Vol 23 (8) ◽

pp. 2083

Author(s):

Dahee Kim ◽

Ja-Il Goo ◽

Mi Kim ◽

Sung-Jin Lee ◽

Moonju Choi ◽

...

Keyword(s):

Hepatitis C Virus ◽

Hepatitis C ◽

Lipid Droplets ◽

Particle Production ◽

Critical Role ◽

Antiviral Effect ◽

Virus Genome ◽

Efficient Production ◽

Genome Replication ◽

Viral Genome Replication

Diacylglycerol acyltransferases (DGATs) play a critical role in the biosynthesis of endogenous triglycerides (TGs) and formation of lipid droplets (LDs) in the liver. In particular, one member of DGATs, DGAT-1 was reported to be an essential host factor for the efficient production of hepatitis C virus (HCV) particles. By utilizing our previously characterized three different groups of twelve DGAT inhibitors, we found that one of the DGAT inhibitors, a 2-((4-adamantylphenoxy) methyl)-N-(furan-2-ylmethyl)-1H-benzo[d]imidazole-5-carboxam (10j) is a potent suppressor of both HCV genome replication and particle production. 10j was able to induce inhibition of these two critical viral functions in a mutually separate manner. Abrogation of the viral genome replication by 10j led to a significant reduction in the viral protein expression as well. Interestingly, we found that its antiviral effect did not depend on the reduction of TG biosynthesis by 10j. This suggests that the inhibitory activity of 10j against DGATs may not be directly related with its antiviral action.

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Nucleotide Analogues as Inhibitors of Viral Polymerases

10.1101/2020.01.30.927574 ◽

2020 ◽

Cited By ~ 12

Author(s):

Jingyue Ju ◽

Shiv Kumar ◽

Xiaoxu Li ◽

Steffen Jockusch ◽

James J. Russo

Keyword(s):

Public Health ◽

Hepatitis C ◽

Viral Genome ◽

Genome Replication ◽

Global Public Health ◽

Nucleotide Analogues ◽

Public Health Emergencies ◽

Polymerase Inhibitors ◽

Viral Genome Replication ◽

Novel Strategy

SummaryCoronaviruses such as the newly discovered virus from Wuhan, China, 2019-nCoV, and the viruses that cause SARS and MERS, have resulted in regional and global public health emergencies. Based on our molecular insight that the hepatitis C virus and the coronavirus use a similar viral genome replication mechanism, we reasoned that the FDA-approved drug EPCLUSA (Sofosbuvir/Velpatasvir) for the treatment of hepatitis C will also inhibit the above coronaviruses, including 2019-nCoV. To develop broad spectrum anti-viral agents, we further describe a novel strategy to design and synthesize viral polymerase inhibitors, by combining the ProTide Prodrug approach used in the development of Sofosbuvir with the use of 3’-blocking groups that we have previously built into nucleotide analogues that function as polymerase terminators.

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