Hepatitis C Viral Replication Complex

Li, Huichun; Yang, Chee-Hing; Lo, Shih-Yen

doi:10.3390/v13030520

Cited by 22 publications

(10 citation statements)

References 221 publications

(205 reference statements)

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“…Assembly of new viral particles is initiated at specific sites of rearranged membrane structures. Such membrane vesicles pass through the Golgi apparatus and are released by exocytosis ( Alazard-Dany et al, 2019 ; Dustin et al, 2016 ; Li et al, 2021 ). Many host biomolecules are exploited during the HCV replication cycle, including cyclophilin A, liver-specific miRNAs, phosphatidylinositol 4-kinase IIIα, diacylglycerol O-acyltransferase 1, and endosomal sorting complexes required for transport (ESCRT) proteins ( Manns et al, 2017 ).…”

Section: Hepatitis C Virus (Hcv) and Antiviral Therapymentioning

confidence: 99%

Viral proteases as therapeutic targets

Majerová

Konvalinka

2022

Molecular Aspects of Medicine

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Section: Hepatitis C Virus (Hcv) and Antiviral Therapymentioning

confidence: 99%

Viral proteases as therapeutic targets

Majerová

Konvalinka

2022

Molecular Aspects of Medicine

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“…2 Institute of Human Virology, Zhongshan School of Medicine, and Key Laboratory of Tropical Disease Control of Ministry of Education, Sun Yat-Sen University, Guangzhou 510080, China. 3 Department of Infectious Diseases, Hainan General Hospital, Hainan Affiliated Hospital of Hainan Medical University, Haikou, China. 4 Department of Infectious Diseases, The Fifth Hospital of Sun Yat-Sen University, Zhuhai, China.…”

Section: Declarations Ethics Approval and Consent To Participatementioning

confidence: 99%

“…HCV NS5B is an RNA dependent RNA polymerase required for viral RNA replication [2]. Numerous host factors have been identified to be involved in and regulate HCV RNA replication by different mechanisms, such as acting on viral proteins, RNA, or the replication complex [3,4]. In general, virus propagation in the cell displays a complex landscape of virus-host interaction and uncovering the host factors involved is critical for understanding the mechanism of HCV RNA replication and viral pathogenesis.…”

Section: Introductionmentioning

confidence: 99%

PPP2R5D promotes hepatitis C virus infection by binding to viral NS5B and enhancing viral RNA replication

Anwar

Zhou

et al. 2022

Virol J

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Background Hepatitis C virus (HCV) infection increased the risk of hepatocellular carcinoma. Identification of host factors required for HCV infection will help to unveil the HCV pathogenesis. Adaptive mutations that enable the replication of HCV infectious clones could provide hints that the mutation-carrying viral protein may specifically interact with some cellular factors essential for the HCV life cycle. Previously, we identified D559G mutation in HCV NS5B (RNA dependent RNA polymerase) important for replication of different genotype clones. Here, we searched for the factors that potentially interacted with NS5B and investigated its roles in HCV infection. Methods Wild-type-NS5B and D559G-NS5B of HCV genotype 2a clone, J6cc, were ectopically expressed in hepatoma Huh7.5 cells, and NS5B-binding proteins were pulled down and identified by mass spectrometry. The necessity and mode of action of the selected cellular protein for HCV infection were explored by experiments including gene knockout or knockdown, complementation, co-immunoprecipitation (Co-IP), colocalization, virus infection and replication, and enzymatic activity, etc. Results Mass spectrometry identified a number of cellular proteins, of which protein phosphatase 2 regulatory subunit B’delta (PPP2R5D, the PP2A regulatory B subunit) was one of D559G-NS5B-pulled down proteins and selected for further investigation. Co-IP confirmed that PPP2R5D specifically interacted with HCV NS5B but not HCV Core and NS3 proteins, and D559G slightly enhanced the interaction. NS5B also colocalized with PPP2R5D in the endoplasmic reticulum. Knockdown and knockout of PPP2R5D decreased and abrogated HCV infection in Huh7.5 cells, respectively, while transient and stable expression of PPP2R5D in PPP2R5D-knockout cells restored HCV infection to a level close to that in wild-type Huh7.5 cells. Replicon assay revealed that PPP2R5D promoted HCV replication, but the phosphatase activity and catalytic subunit of PP2A were not affected by NS5B. Conclusions PPP2R5D interactes with HCV NS5B and is required for HCV infection in cultured hepatoma cells through facilitating HCV replication.

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“…HCV NS5B is an RNA dependent RNA polymerase (RdRp) required for viral RNA replication (2). Many host factors have been identi ed to be involved in and regulate HCV RNA replication by different mechanisms, such as disrupting viral proteins, RNA, or the replication complex (3,4). In general, virus propagation in the cell displays a landscape of complex virus-host interaction, and uncovering host factors involved in the interactions is critical to understand the mechanism of HCV RNA replication and viral pathogenesis.…”

Section: Introductionmentioning

confidence: 99%

PPP2R5D Promotes Hepatitis C Virus Infection Through Binding to NS5B Protein

Anwar

Zhou

et al. 2021

Preprint

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Background: Hepatitis C virus (HCV) is an important human pathogen causing chronic hepatitis C, end-stage liver diseases, and hepatocellular carcinoma. The development of infectious HCV cell culture systems primarily relied on the replication enhancement effect of adaptive mutations. Although the mode of action may vary, those adaptive mutations could direct the study of virus-host interactions required for efficient virus infection. We previously identified a substitution D559G in NS5B (RNA dependent RNA polymerase) critical for the replication of HCV genomes. In this study, we set out to study whether D559G-NS5B specifically interacted with some host factors crucial for HCV infection.Methods: Through mass spectrometry analysis of immunoprecipitation mixture of ectopically expressed wild-type and D559G-mutated NS5B, we identified candidate factors showing potential interactions with NS5B and D559G-NS5B. The requirement of selected host factor in HCV infection in vitro was demonstrated by gene knockout, overexpression, virus infection, and co-immunoprecipitation approaches.Results: From the results of immunoprecipitation and mass spectrometry analysis, we selected protein phosphatase 2 regulatory subunit B’delta (PPP2R5D) for further characterization. Co-immunoprecipitation confirmed that both wild-type and D559G NS5B proteins interacted with PPP2R5D, but the interaction between D559G-NS5B and PPP2R5D was more efficient. Silencing of PPP2R5D decreased HCV infection, and knockout of PPP2R5D nearly eliminated HCV infection in Huh7.5 cells. Transient and stably overexpression of PPP2R5D in PPP2R5D-knockout cells restored HCV infection to a level close to that seen for wild-type Huh7.5 cells. Conclusions: PPP2R5D is required for HCV infection in cultured hepatoma cells, and PPP2R5D may function through binding to HCV NS5B. The underlying mechanism of PPP2R5D in the complete HCV life cycle requires further investigation.

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Hepatitis C Viral Replication Complex

Cited by 22 publications

References 221 publications

Viral proteases as therapeutic targets

Viral proteases as therapeutic targets

PPP2R5D promotes hepatitis C virus infection by binding to viral NS5B and enhancing viral RNA replication

PPP2R5D Promotes Hepatitis C Virus Infection Through Binding to NS5B Protein

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