Nuclear receptor 4 group A member 1 determines hepatitis C virus entry efficiency through the regulation of cellular receptor and apolipoprotein E expression

Zhu, Wandi; Pei, Rongjuan; Jin, Rui; Zhou, Yuan; Wang, Yun; Wu, Chunchen; Lu, Mengji; Chen, Xinwen

doi:10.1099/vir.0.065003-0

Cited by 5 publications

(10 citation statements)

References 69 publications

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“…The mRNA and HCV RNA levels were normalized against the copy number of human beta-actin mRNAs. The forward and reverse primers used to amplify PLA1A were 5=-GGAACTGAGAAACAA GGACACC-3= and 5=-AAACTCGGTTGGAAGACTGAAA-3=, and the primers for HCV and actin were previously described (36).…”

Section: Methodsmentioning

confidence: 99%

Phosphatidylserine-Specific Phospholipase A1 Involved in Hepatitis C Virus Assembly through NS2 Complex Formation

Guo

Pei

Yang

et al. 2015

J Virol

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Several members of the phospholipase family have been reported to be involved in hepatitis C virus (HCV) replication. Here, we identified another phospholipase, phosphatidylserine-specific phospholipase A1 (PLA1A), as a host factor involved in HCV assembly. PLA1A was upregulated by HCV infection, and PLA1A knockdown significantly reduced J399EM (genotype 2a) HCV propagation at the assembly step but not the entry, RNA replication, and protein translation steps of the life cycle. Protein localization and interaction analysis further revealed a role of PLA1A in the interaction of NS2-E2 and NS2-NS5A, as the formation of the NS2-E2 and NS2-NS5A complexes was weakened in the absence of PLA1A. In addition, PLA1A stabilized the NS2/NS5A dotted structure during infection. These data suggest that PLA1A plays an important role in bridging the membrane-associated NS2-E2 complex and the NS5A-associated replication complex via its interaction with E2, NS2, and NS5A, which leads to a coordinating interaction between the structural and nonstructural proteins and facilitates viral assembly. IMPORTANCEHepatitis C virus (HCV) genomic replication is driven by the replication complex and occurs at the membranous web, while the lipid droplet is the organelle in which virion assembly is initiated. In this study, we identified phosphatidylserine-specific phospholipase A1 (PLA1A), a member of phospholipase A 1 family, as a novel host factor involved in the assembly process of HCV. PLA1A, which is induced by HCV infection at a late infection stage, interacts with HCV E2, NS2, and NS5A proteins and enhances and stabilizes the NS2-E2 and NS2-NS5A complex formation, which is essential for viral assembly. Thus, PLA1A is an important host factor which is involved in the initiation of the viral assembly in close proximity to Core-decorated lipid droplets through bringing together the HCV replication complex and envelope complex. Hepatitis C virus (HCV) is a major cause of chronic liver disease, affecting approximately 185 million people worldwide (1). HCV is a positive single-stranded RNA virus belonging to the Flaviviridae family. The HCV 9.6-kb genome contains a large open reading frame encoding a single polyprotein that is processed into its structural proteins (Core, E1, and E2) and nonstructural (NS) proteins (p7, NS2, NS3, NS4A, NS4B, NS5A, and NS5B) by host and viral proteinases (2). The structural proteins are components of the virion, while nonstructural proteins NS3 to NS5B compose the minimal viral replicase governing RNA replication (3, 4).The overall HCV life cycle has been well defined since the development of an infectious HCV cell culture system (5-7). HCV genomic replication is driven by the replication complex (RC) and occurs at the membranous web, a rearranged membrane structure induced by virus infection (8-10). Recent progress regarding the study of the assembly process demonstrates that the lipid droplet (LD) is the organelle in which virion assembly is initiated (11) and that, in addition to the structural prote...

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Section: Methodsmentioning

confidence: 99%

Phosphatidylserine-Specific Phospholipase A1 Involved in Hepatitis C Virus Assembly through NS2 Complex Formation

Guo

Pei

Yang

et al. 2015

J Virol

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“…The virus in the supernatant was concentrated and purified by PEG 8,000 and ultracentrifugation sequentially, and then titered by immunofluorescence staining with NS3 antibody 8 G-2 (Abcam®, ab65407) (Jones et al, 2011 ). The J399EM virus which is derived from the JFH-1 virus (kindly provided by Takaji Wakita) by inserting EGFP into the NS5A, was prepared as previously described (Zhu et al, 2014 ). The HCV pseudoparticle (HCVpp) was generated by transfection of 293T cells with pNL4.3.lucR-E- and pcDNA3.1-E1E2 (a gift from Jin Zhong) plasmids (Zhu et al, 2014 ).…”

Section: Methodsmentioning

confidence: 99%

“…The J399EM virus which is derived from the JFH-1 virus (kindly provided by Takaji Wakita) by inserting EGFP into the NS5A, was prepared as previously described (Zhu et al, 2014 ). The HCV pseudoparticle (HCVpp) was generated by transfection of 293T cells with pNL4.3.lucR-E- and pcDNA3.1-E1E2 (a gift from Jin Zhong) plasmids (Zhu et al, 2014 ).…”

Section: Methodsmentioning

confidence: 99%

RNA binding protein 24 regulates the translation and replication of hepatitis C virus

et al. 2018

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The secondary structures of hepatitis C virus (HCV) RNA and the cellular proteins that bind to them are important for modulating both translation and RNA replication. However, the sets of RNA-binding proteins involved in the regulation of HCV translation, replication and encapsidation remain unknown. Here, we identified RNA binding motif protein 24 (RBM24) as a host factor participated in HCV translation and replication. Knockdown of RBM24 reduced HCV propagation in Huh7.5.1 cells. An enhanced translation and delayed RNA synthesis during the early phase of infection was observed in RBM24 silencing cells. However, both overexpression of RBM24 and recombinant human RBM24 protein suppressed HCV IRES-mediated translation. Further analysis revealed that the assembly of the 80S ribosome on the HCV IRES was interrupted by RBM24 protein through binding to the 5′-UTR. RBM24 could also interact with HCV Core and enhance the interaction of Core and 5′-UTR, which suppresses the expression of HCV. Moreover, RBM24 enhanced the interaction between the 5′- and 3′-UTRs in the HCV genome, which probably explained its requirement in HCV genome replication. Therefore, RBM24 is a novel host factor involved in HCV replication and may function at the switch from translation to replication.

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“…Total RNA was isolated with the TRIzol or TRIzol LS reagent (Invitrogen, Carlsbad, California, USA), and real-time RT-PCR was performed using a QuantiFast SYBR green RT-PCR kit (Qiagen, Hilden, Germany) as previously described [ 31 ]. The following primers were used for the real-time RT-PCR: PIAS2 sense: 5′-CTCATCAAGCCCACGAGTTTAG-3′ and antisense: 5′-CCAGGCAAAGTCTCAACTGAA-3′; HCV sense: 5′-ATCACTCCCCTGTGAGGAACT-3′ and antisense: 5′-GCGGGTTGATCCAAGAAAGG-3′; the primers for actin were previously described [ 33 ].…”

Section: Methodsmentioning

confidence: 99%

Protein Inhibitor of Activated STAT2 Restricts HCV Replication by Modulating Viral Proteins Degradation

Guo

Chen

Gao

et al. 2017

Viruses

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Hepatitis C virus (HCV) replication in cells is controlled by many host factors. In this report, we found that protein inhibitor of activated STAT2 (PIAS2), which is a small ubiquitin-like modifier (SUMO) E3 ligase, restricted HCV replication. During infection, HCV core, NS3 and NS5A protein expression, as well as the viral assembly and budding efficiency were enhanced when endogenous PIAS2 was knocked down, whereas exogenous PIAS2 expression decreased HCV core, NS3, and NS5A protein expression and the viral assembly and budding efficiency. PIAS2 did not influence the viral entry, RNA replication, and protein translation steps of the viral life cycle. When expressed together with SUMO1, PIAS2 reduced the HCV core, NS3 and NS5A protein levels expressed from individual plasmids through the proteasome pathway in a ubiquitin-independent manner; the stability of these proteins in the HCV infectious system was enhanced when PIAS2 was knocked down. Furthermore, we found that the core was SUMOylated at amino acid K78, and PIAS2 enhanced the SUMOylation level of the core.

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Nuclear receptor 4 group A member 1 determines hepatitis C virus entry efficiency through the regulation of cellular receptor and apolipoprotein E expression

Cited by 5 publications

References 69 publications

Phosphatidylserine-Specific Phospholipase A1 Involved in Hepatitis C Virus Assembly through NS2 Complex Formation

Phosphatidylserine-Specific Phospholipase A1 Involved in Hepatitis C Virus Assembly through NS2 Complex Formation

RNA binding protein 24 regulates the translation and replication of hepatitis C virus

Protein Inhibitor of Activated STAT2 Restricts HCV Replication by Modulating Viral Proteins Degradation

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