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

Phongphaew, Wallaya; Kobayashi, Shintaro; Sasaki, Michihito; Carr, Michael J.; Hall, William W.; Orba, Yasuko; Sawa, Hirofumi

doi:10.1016/j.virusres.2016.11.029

Cited by 30 publications

(32 citation statements)

References 81 publications

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“…Although we have illustrated free nucleocapsids within the cytosol, we cannot exclude the possibility that fusion is tightly coupled to capsid ubiquitylation and disassembly, such that nucleocapsids may be ubiquitylated and disassembled as they are exposed to the cytosol. Consistent with our model, VCP/p97 activity was previously shown to be important for WNV, JEV, and DENV infection, although specific role(s) for VCP/p97 in virus entry were not determined (53,83). Similarly, VCP/p97 is also important for an early step, upstream of N-protein degradation, during the entry of infectious bronchitis virus, a coronavirus (84).…”

Section: Discussionsupporting

confidence: 82%

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: Discussionsupporting

confidence: 82%

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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“…Mutations in the sequences of the C protein or structural proteins of WNV were introduced via inverse PCR. The plasmid expressing envelope protein prME (pCXSN-prME) and the plasmid expressing the subgenomic RNA of WNV (pCMV-WNrep-DsRed) were previously constructed [18,49]. The plasmid expressing the NS proteins (pWNIIrep-GFP, was generously provided by Dr. Robert W. Doms) was previously constructed [50].…”

Section: Plasmid Constructionmentioning

confidence: 99%

“…Rabbit anti-JEV serum that exhibited cross-reactivity with the WNV E protein was produced as previously described [51]. Rabbit anti-NS3 serum was prepared as previously described [49]. The following antibodies were purchased: mouse anti-WNV E protein monoclonal antibody (Merck Millipore, Billerica, MA, USA); mouse anti-ubiquitin monoclonal antibody (Enzo Life Sciences Inc., Farmingdale, NY, USA); mouse anti-actin monoclonal antibody (Wako); mouse anti-LC3 monoclonal antibody, rabbit anti-DDDDK-tag, anti-LC3 and anti-p62 polyclonal antibodies (MBL, Nagoya, Japan); and rabbit anti-AMPK polyclonal and monoclonal antibodies (Cell Signaling Technology, Beverly, MA, USA).…”

Section: Antibodies Reagents and Transfectionmentioning

confidence: 99%

West Nile virus capsid protein inhibits autophagy by AMP-activated protein kinase degradation in neurological disease development

et al. 2020

Self Cite

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West Nile virus (WNV) belongs to the Flaviviridae family and has emerged as a significant cause of viral encephalitis in birds and animals including humans. WNV replication directly induces neuronal injury, followed by neuronal cell death. We previously showed that accumulation of ubiquitinated protein aggregates was involved in neuronal cell death in the WNV-infected mouse brain. In this study, we attempted to elucidate the mechanisms of the accumulation of protein aggregates in the WNV-infected cells. To identify the viral factor inducing the accumulation of ubiquitinated proteins, intracellular accumulation of ubiquitinated proteins was examined in the cells expressing the viral protein. Expression of capsid (C) protein induced the accumulation, while mutations at residues L51 and A52 in C protein abrogated the accumulation. Wild-type (WT) or mutant WNV in which mutations were introduced into the residues was inoculated into human neuroblastoma cells. The expression levels of LC3-II, an autophagy-related protein, and AMP-activated protein kinase (AMPK), an autophagy inducer, were reduced in the cells infected with WT WNV, while the reduction was not observed in the cells infected with WNV with the mutations in C protein. Similarly, ubiquitination and degradation of AMPK were only observed in the cells infected with WT WNV. In the cells expressing C protein, AMPK was co-precipitated with C protein and mutations in L51 and A52 reduced the interaction. Although the viral replication was not affected, the accumulation of ubiquitinated proteins in brain and neurological symptoms were attenuated in the mouse inoculated with WNV with the mutations in C protein as compared with that with WT WNV. Taken together, ubiquitination and degradation of AMPK by C protein resulted in the inhibition of autophagy and the accumulation of protein aggregates, which contributes to the development of neurological disease.

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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%

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

Cited by 30 publications

References 81 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

West Nile virus capsid protein inhibits autophagy by AMP-activated protein kinase degradation in neurological disease development

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

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