Affinity Purification of the Hepatitis C Virus Replicase Identifies Valosin-Containing Protein, a Member of the ATPases Associated with Diverse Cellular Activities Family, as an Active Virus Replication Modulator

Yi, Zhigang; Fang, Changming; Zou, Jingyi; Xu, Jun; Song, Wuhui; Du, Xiaoting; Pan, Tingting; Lu, Haojie; Yuan, Zhenghong

doi:10.1128/jvi.01140-16

Cited by 28 publications

(38 citation statements)

References 71 publications

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“…VCP/p97 also contributes to productive trafficking of NRAMP2, a cellular receptor for the entry of Sindbis virus, an alphavirus (85). Thus, VCP/p97 appears to play a general role in the entry of enveloped RNA viruses and may also contribute to RNA replication of alphaviruses (86) and hepaciviruses (87,88). Given these findings, it will be interesting to determine whether VCP/p97 inhibitors have therapeutic potential in viral infections.…”

Section: Discussionmentioning

confidence: 99%

A Sensitive Yellow Fever Virus Entry Reporter Identifies Valosin-Containing Protein (VCP/p97) as an Essential Host Factor for Flavivirus Uncoating

Ramanathan

Zhang

Douam

et al. 2020

mBio

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While the basic mechanisms of flavivirus entry and fusion are understood, little is known about the postfusion events that precede RNA replication, such as nucleocapsid disassembly. We describe here a sensitive, conditionally replicationdefective yellow fever virus (YFV) entry reporter, YFVΔSK/Nluc, to quantitively monitor the translation of incoming, virus particle-delivered genomes. We validated that YFVΔSK/Nluc gene expression can be neutralized by YFV-specific antisera and requires known flavivirus entry pathways and cellular factors, including clathrin-and dynamin-mediated endocytosis, endosomal acidification, YFV E glycoprotein-mediated fusion, and cellular LY6E and RPLP1 expression. The initial round of YFV translation was shown to require cellular ubiquitylation, consistent with recent findings that dengue virus capsid protein must be ubiquitylated in order for nucleocapsid uncoating to occur. Importantly, translation of incoming YFV genomes also required valosin-containing protein (VCP)/p97, a cellular ATPase that unfolds and extracts ubiquitylated client proteins from large complexes. RNA transfection and washout experiments showed that VCP/p97 functions at a postfusion, pretranslation step in YFV entry. Finally, VCP/p97 activity was required by other flaviviruses in mammalian cells and by YFV in mosquito cells. Together, these data support a critical role for VCP/p97 in the disassembly of incoming flavivirus nucleocapsids during a postfusion step in virus entry. IMPORTANCE Flaviviruses are an important group of RNA viruses that cause significant human disease. The mechanisms by which flavivirus nucleocapsids are disassembled during virus entry remain unclear. Here, we used a yellow fever virus entry reporter, which expresses a sensitive reporter enzyme but does not replicate, to show that nucleocapsid disassembly requires the cellular protein-disaggregating enzyme valosin-containing protein, also known as p97.

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Section: Discussionmentioning

confidence: 99%

A Sensitive Yellow Fever Virus Entry Reporter Identifies Valosin-Containing Protein (VCP/p97) as an Essential Host Factor for Flavivirus Uncoating

Ramanathan

Zhang

Douam

et al. 2020

mBio

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“…The possible mechanism(s) of VCP involvement in WNV infection may be based on the localization and physiological function of VCP. Functional roles of VCP in the replication of the WNV-related virus in the family Flaviviridae, hepatitis C virus (HCV) have been reported (Yi et al, 2016). VCP knockdown significantly decreased expression of HCV RNA levels and VCP was found to be colocalized with the HCV replication complex.…”

Section: Discussionmentioning

confidence: 99%

Valosin-containing protein (VCP/p97) plays a role in the replication of West Nile virus

et al. 2017

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Valosin-containing protein (VCP) is classified as a member of the type II AAA(+) ATPase protein family. VCP functions in several cellular processes, including protein degradation, membrane fusion, vesicular trafficking and disassembly of stress granules. Moreover, VCP is considered to play a role in the replication of several viruses, albeit through different mechanisms. In the present study, we have investigated the role of VCP in West Nile virus (WNV) infection. Endogenous VCP expression was inhibited using either VCP inhibitors or by siRNA knockdown. It could be shown that the inhibition of endogenous VCP expression significantly inhibited WNV infection. The entry assay revealed that silencing of endogenous VCP caused a significant reduction in the expression levels of WNV-RNA compared to control siRNA-treated cells. This indicates that VCP may play a role in early steps either the binding or entry steps of the WNV life cycle. Using WNV virus like particles and WNV-DNA-based replicon, it could be demonstrated that perturbation of VCP expression decreased levels of newly synthesized WNV genomic RNA. These findings suggest that VCP is involved in early steps and during genome replication of the WNV life cycle. (C) 2016 Elsevier B.V. All rights reserved

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“…reported that VCP participated in viral genome replication of poliovirus, hepatitis C virus and West Nile virus (Arita et al, 2012;Phongphaew et al, 2017;Yi et al, 2016). In addition, roles of VCP in early stages of viral infection have been reported for Coronavirus and Sindbis virus (Panda et al, 2013;Wong et al, 2015).…”

Section: Discussionmentioning

confidence: 99%

“…VCP contains two ATP-binding (AAA) domains. Double mutation (E305Q/E578Q) of these two domains completely abolishes ATPase activity, as a dominant negative mutant (Yi et al, 2016). To determine whether the ATPase activity of VCP is required for EVA71 replication, we performed a rescue experiment by co-transfection of shRNA against VCP and overexpression of shRNA-resistant VCP (VCP-es) or dominant negative mutant VCP (VCP-es-DN).…”

Section: The Atpase Activity Of Vcp Is Required In Eva71 Infectionmentioning

confidence: 99%

“…Valosin-containing protein (VCP), a hexameric type II AAA ATPase, participates in various cellular activities including protein homeostasis, DNA replication and repair and autophagy (Meyer and Weihl, 2014). VCP has been found to play roles in replication of poliovirus and Hepatitis C virus (Arita et al, 2012;Yi et al, 2016), and originally identified as host factor of Drosophila C virus (Cherry et al, 2006). Recently, VCP was identified to be required in EVA71 replication (Wu et al, 2016) and VCP co-exists with the viral protein and other known replication-related molecules in EVA71-induced ROs (Wang et al, 2017).…”

Section: Introductionmentioning

confidence: 99%

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Involvement of VCP/UFD1/Nucleolin in the viral entry of Enterovirus A species

Yan

Wang

et al. 2020

Virus Research

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A B S T R A C TValosin-containing protein (VCP) plays roles in various cellular activities. Recently, Enterovirus A71 (EVA71) infection was found to hijack the VCP protein. However, the mechanism by which VCP participates in the EVA71 life cycle remains unclear. Using chemical inhibitor, RNA interference and dominant negative mutant, we confirmed that the VCP and its ATPase activity were critical for EVA71 infection. To identify the factors downstream of VCP in enterovirus infection, 31 known VCP-cofactors were screened in the siRNA knockdown experiments. The results showed that UFD1 (ubiquitin recognition factor in ER associated degradation 1), but not NPL4 (NPL4 homolog, ubiquitin recognition factor), played critical roles in infections by EVA71. UFD1 knockdown suppressed the activity of EVA71 pseudovirus (causing single round infection) while it did not affect the viral replication in replicon RNA transfection assays. In addition, knockdown of VCP and UFD1 reduced viral infections by multiple human Enterovirus A serotypes. Mechanistically, we found that knockdown of UFD1 significantly decreased the binding and the subsequent entry of EVA71 to host cells through modulating the levels of nucleolin protein, a coreceptor of EVA71. Together, these data reveal novel roles of VCP and its cofactor UFD1 in the virus entry by EVA71. 3A-3D) . The viral RNA-dependent RNA polymerase https://doi.

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Affinity Purification of the Hepatitis C Virus Replicase Identifies Valosin-Containing Protein, a Member of the ATPases Associated with Diverse Cellular Activities Family, as an Active Virus Replication Modulator

Cited by 28 publications

References 71 publications

A Sensitive Yellow Fever Virus Entry Reporter Identifies Valosin-Containing Protein (VCP/p97) as an Essential Host Factor for Flavivirus Uncoating

A Sensitive Yellow Fever Virus Entry Reporter Identifies Valosin-Containing Protein (VCP/p97) as an Essential Host Factor for Flavivirus Uncoating

Valosin-containing protein (VCP/p97) plays a role in the replication of West Nile virus

Involvement of VCP/UFD1/Nucleolin in the viral entry of Enterovirus A species

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