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

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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Insertion and deletion analyses identify regions of non-structural protein 5A of Hepatitis C virus that are dispensable for viral genome replication

Journal of General Virology ◽

10.1099/vir.0.81407-0 ◽

2006 ◽

Vol 87 (2) ◽

pp. 323-327 ◽

Cited By ~ 23

Author(s):

Shuanghu Liu ◽

Israrul H. Ansari ◽

Subash C. Das ◽

Asit K. Pattnaik

Keyword(s):

Hepatitis C Virus ◽

Hepatitis C ◽

Viral Genome ◽

Fluorescent Protein ◽

Genome Replication ◽

Structural Protein ◽

Mutant Proteins ◽

Viral Genome Replication ◽

Green Fluorescent ◽

Carboxy Terminal

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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Alternative translation and alternative RNA processing mechanism in parvovirus RNA processing

10.32469/10355/45905 ◽

2014 ◽

Author(s):

◽

Olufemi Fasina

Keyword(s):

Gene Expression ◽

Rna Processing ◽

Viral Genome ◽

Transcription Initiation ◽

Alternative Polyadenylation ◽

Open Reading Frames ◽

Genome Replication ◽

University Of Missouri ◽

Diverse Range ◽

Viral Genome Replication

[ACCESS RESTRICTED TO THE UNIVERSITY OF MISSOURI AT AUTHOR'S REQUEST.] Viruses as obligate intracellular metabolic parasite require the capacity to orchestrate and modulate the host environment either in the nucleus or cytoplasm for their efficient reproductive life cycle. This warrants the use of diverse range of proteins expressed from the viral genome with the ability of regulating viral genome replication, transcription and translation, in addition antagonizing host factors inhibitory to the virus. Therefore, in order to achieve these goals, viruses utilizes gene expression strategies to expand their coding capacity. Gene expression mechanism such as transcription initiation, capping, splicing and 3ï¿½-end processing afford viruses the opportunities to utilize the eukaryotic metabolic machineries for generating proteome diversity. Parvoviruses and other DNA viruses effectively capitalize on their use of nuclear eukaryotic metabolic machineries to co-opt host cell factors for optimal replication and gene expression. Parvoviruses with small genome size and overlapping open reading frames utilize alternative transcription initiation, alternative splicing and alternative polyadenylation to co-ordinate the expression of its non-structural and structural proteins. In this work, we have characterized how two parvoviruses; Dependovirus AAV5 and Bocavirus Minute virus of canine (MVC) utilize alternative gene expression mechanisms and strategies to optimize expression of viral proteins from their genome.

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Dimerisation of the influenza virus RNA polymerase during viral genome replication

Access Microbiology ◽

10.1099/acmi.ac2019.po0145 ◽

2019 ◽

Vol 1 (1A) ◽

Author(s):

Alexander Walker ◽

Haitian Fan ◽

Loic Carrique ◽

Jeremy Keown ◽

David Bauer ◽

...

Keyword(s):

Influenza Virus ◽

Rna Polymerase ◽

Viral Genome ◽

Genome Replication ◽

Virus Rna ◽

Viral Genome Replication

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The formation and modification of chromatin-like structure of human parvovirus B19 regulate viral genome replication and RNA processing

Virus Research ◽

10.1016/j.virusres.2017.03.001 ◽

2017 ◽

Vol 232 ◽

pp. 134-138 ◽

Cited By ~ 1

Author(s):

Huanzhou Xu ◽

Sujuan Hao ◽

Junmei Zhang ◽

Zhen Chen ◽

Hanzhong Wang ◽

...

Keyword(s):

Rna Processing ◽

Viral Genome ◽

Parvovirus B19 ◽

Genome Replication ◽

Human Parvovirus B19 ◽

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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Mosquito metabolomics reveal that dengue virus replication requires phospholipid reconfiguration via the remodeling cycle

Proceedings of the National Academy of Sciences ◽

10.1073/pnas.2015095117 ◽

2020 ◽

Vol 117 (44) ◽

pp. 27627-27636

Author(s):

Thomas Vial ◽

Wei-Lian Tan ◽

Eric Deharo ◽

Dorothée Missé ◽

Guillaume Marti ◽

...

Keyword(s):

Dengue Virus ◽

Viral Genome ◽

De Novo ◽

Genome Replication ◽

Mosquito Vector ◽

Phospholipid Biosynthesis ◽

Replication Complex ◽

Mosquito Cells ◽

Viral Genome Replication ◽

Blood Meals

Dengue virus (DENV) subdues cell membranes for its cellular cycle by reconfiguring phospholipids in humans and mosquitoes. Here, we determined how and why DENV reconfigures phospholipids in the mosquito vector. By inhibiting and activating the de novo phospholipid biosynthesis, we demonstrated the antiviral impact of de novo–produced phospholipids. In line with the virus hijacking lipids for its benefit, metabolomics analyses indicated that DENV actively inhibited the de novo phospholipid pathway and instead triggered phospholipid remodeling. We demonstrated the early induction of remodeling during infection by using isotope tracing in mosquito cells. We then confirmed in mosquitoes the antiviral impact of de novo phospholipids by supplementing infectious blood meals with a de novo phospholipid precursor. Eventually, we determined that phospholipid reconfiguration was required for viral genome replication but not for the other steps of the virus cellular cycle. Overall, we now propose that DENV reconfigures phospholipids through the remodeling cycle to modify the endomembrane and facilitate formation of the replication complex. Furthermore, our study identified de novo phospholipid precursor as a blood determinant of DENV human-to-mosquito transmission.

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