A Map of the Arenavirus Nucleoprotein-Host Protein Interactome Reveals that Junín Virus Selectively Impairs the Antiviral Activity of Double-Stranded RNA-Activated Protein Kinase (PKR)

King, Benjamin R.; Hershkowitz, Dylan C; Eisenhauer, Philip; Weir, Marion E.; Ziegler, Christopher M.; Russo, Joanne; Bruce, Emily A.; Ballif, Bryan A.; Botten, Jason

doi:10.1128/jvi.00763-17

Cited by 42 publications

(87 citation statements)

References 56 publications

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“…In conclusion, we have provided a detailed map of JUNV virus interactions with the human proteome and identified several host factors required for JUNV infection. This work, combined with our previous proteomics studies (74,96), represents a crucial step in understanding the protein networks hijacked by arenaviruses to drive the virus life cycle. This study highlights the important role that the ESCRT pathway plays in driving infectious JUNV particle release, while further demonstrating the diversity exhibited among arenaviruses in their use of ESCRT machinery for the production of different classes of virus particles.…”

Section: Discussionmentioning

confidence: 89%

A Proteomics Survey of Junín Virus Interactions with Human Proteins Reveals Host Factors Required for Arenavirus Replication

Ziegler

Eisenhauer

Kelly

et al. 2018

J Virol

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Arenaviruses are negative-strand, enveloped RNA viruses that cause significant human disease. In particular, Junín mammarenvirus (JUNV) is the etiologic agent of Argentine hemorrhagic fever. At present, little is known about the cellular proteins that the arenavirus matrix protein (Z) hijacks to accomplish its various functions, including driving the process of virus release. Further, there is a little knowledge regarding host proteins incorporated into arenavirus particles and their importance for virion function. To address these deficiencies, we used mass spectrometry to identify human proteins that (i) interact with the JUNV matrix protein inside of cells or within virus-like particles (VLPs) and/or (ii) are incorporated into JUNV strain Candid #1 particles. Bioinformatic analyses revealed that multiple classes of human proteins were overrepresented in the datasets, including ribosomal proteins, Ras superfamily proteins, and endosomal sorting complex required for transport (ESCRT) proteins. Several of these proteins were required for the propagation of JUNV (ARF1, ATP6V0D1 and PRDX3), lymphocytic choriomeningitis mammarenavirus (LCMV) (Rab5c), or both viruses (ATP5B, IMPDH2). Further, we show that release of infectious JUNV particles, but not LCMV particles, requires a functional ESCRT pathway and that ATP5B and IMPDH2 are required for JUNV budding. In summary, we have provided a large-scale map of host machinery that associates with JUNV and identified key human proteins required for its propagation. This dataset provides a resource for the field to guide antiviral target discovery and to better understand the biology of the arenavirus matrix protein and the importance of host proteins for virion function. Arenaviruses are deadly human pathogens for which there are no United States Food and Drug Administration-approved vaccines and only limited treatment options. Little is known about the host proteins that are incorporated into arenavirus particles or that associate with its multifunctional matrix protein. Using Junín mammarenavirus (JUNV), the causative agent of Argentine hemorrhagic fever, as a model organism, we mapped the human proteins that are incorporated into JUNV particles or that associate with the JUNV matrix protein. Functional analysis revealed host machinery that is required for JUNV propagation, including the cellular ESCRT pathway. This study improves our understanding of critical arenavirus-host interactions and provides a dataset that will guide future studies to better understand arenavirus pathogenesis and identify novel host proteins that can be therapeutically targeted.

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Section: Discussionmentioning

confidence: 89%

A Proteomics Survey of Junín Virus Interactions with Human Proteins Reveals Host Factors Required for Arenavirus Replication

Ziegler

Eisenhauer

Kelly

et al. 2018

J Virol

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“…While our manuscript was in preparation, King et al reported that infection with the Candid#1 vaccine strain of JUNV activated the PKR response in infected cells and that small interfering RNA (siRNA)-mediated knockdown of PKR had no impact on viral propagation (50). Furthermore, JUNV NP is shown to interact with PKR in infected cells, suggesting that JUNV NP might locally inhibit PKR activity, thereby facilitating viral protein synthesis.…”

Section: Discussionmentioning

confidence: 93%

“…These results suggest a phenotypic difference in the impact of virus infection on protein synthesis between the vaccine and pathogenic strains of JUNV, which remains to be investigated in future studies. Another potential way to explain the observed differences is that we prepared and used recombinant virus stocks that are likely less heterogeneous than the vaccine Candid#1 strain used in the aforementioned study (50).…”

Section: Discussionmentioning

confidence: 99%

Highly Pathogenic New World Arenavirus Infection Activates the Pattern Recognition Receptor Protein Kinase R without Attenuating Virus Replication in Human Cells

Huang

Kolokoltsova

Mateer

et al. 2017

J Virol

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The arenavirus family consists of several highly pathogenic viruses, including the Old World (OW) arenavirus Lassa fever virus (LASV) and the New World (NW) Junin virus (JUNV) and Machupo virus (MACV). Host response to infection by these pathogenic arenaviruses is distinct in many aspects. JUNV and MACV infections readily induce an interferon (IFN) response in human cells, while LASV infection usually triggers an undetectable or weak IFN response. JUNV induces an IFN response through RIG-I, suggesting that the host non-self RNA sensor readily detects JUNV viral RNAs (vRNAs) during infection and activates IFN response. Double-stranded-RNA (dsRNA)-activated protein kinase R (PKR) is another host non-self RNA sensor classically known for its vRNA recognition activity. Here we report that infection with NW arenaviruses JUNV and MACV, but not OW LASV, activated PKR, concomitant with elevated phosphorylation of the translation initiation factor α subunit of eukaryotic initiation factor 2 (eIF2α). Host protein synthesis was substantially suppressed in MACV- and JUNV-infected cells but was only marginally reduced in LASV-infected cells. Despite the antiviral activity known for PKR against many other viruses, the replication of JUNV and MACV was not impaired but was slightly augmented in wild-type (wt) cells compared to that in PKR-deficient cells, suggesting that PKR or PKR activation did not negatively affect JUNV and MACV infection. Additionally, we found an enhanced IFN response in JUNV- or MACV-infected PKR-deficient cells, which was inversely correlated with virus replication. The detection of viral RNA by host non-self RNA sensors, including RIG-I and MDA5, is critical to the initiation of the innate immune response to RNA virus infection. Among pathogenic arenaviruses, the OW LASV usually does not elicit an interferon response. However, the NW arenaviruses JUNV and MACV readily trigger an IFN response in a RIG-I-dependent manner. Here, we demonstrate for the first time that pathogenic NW arenaviruses JUNV and MACV, but not the OW arenavirus LASV, activated the dsRNA-dependent PKR, another host non-self RNA sensor, during infection. Interestingly, the replication of JUNV and MACV was not restricted but was rather slightly augmented in the presence of PKR. Our data provide new evidence for a distinct interplay between host non-self RNA sensors and pathogenic arenaviruses, which also provides insights into the pathogenesis of arenaviruses and may facilitate the design of vaccines and treatments against arenavirus-caused diseases.

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“…To date, lists of host-cell proteins associated with various arenavirus infections have been established, providing new bases to investigate the potential roles of such interactions during the course of infection. With the exception of one study, which focused on the proteins of cellular origin associated with the Z protein of JUNV, most reports have focused on the NP and L interactomes during JUNV and LCMV infection [44][45][46][47][48]. However, these reports were based on the expression of tagged viral proteins through transfection or infection with genetically modified viruses and the identification of associated host-cell partners after purification and mass spectrometry analysis.…”

Section: Discussionmentioning

confidence: 99%

E3 Ligase ITCH Interacts with the Z Matrix Protein of Lassa and Mopeia Viruses and Is Required for the Release of Infectious Particles

Baillet

Krieger

Carnec

et al. 2019

Viruses

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Lassa virus (LASV) and Mopeia virus (MOPV) are two closely related, rodent-born mammarenaviruses. LASV is the causative agent of Lassa fever, a deadly hemorrhagic fever endemic in West Africa, whereas MOPV is non-pathogenic in humans. The Z matrix protein of arenaviruses is essential to virus assembly and budding by recruiting host factors, a mechanism that remains partially defined. To better characterize the interactions involved, a yeast two-hybrid screen was conducted using the Z proteins from LASV and MOPV as a bait. The cellular proteins ITCH and WWP1, two members of the Nedd4 family of HECT E3 ubiquitin ligases, were found to bind the Z proteins of LASV, MOPV and other arenaviruses. The PPxY late-domain motif of the Z proteins is required for the interaction with ITCH, although the E3 ubiquitin-ligase activity of ITCH is not involved in Z ubiquitination. The silencing of ITCH was shown to affect the replication of the old-world mammarenaviruses LASV, MOPV, Lymphocytic choriomeningitis virus (LCMV) and to a lesser extent Lujo virus (LUJV). More precisely, ITCH was involved in the egress of virus-like particles and the release of infectious progeny viruses. Thus, ITCH constitutes a novel interactor of LASV and MOPV Z proteins that is involved in virus assembly and release.Viruses 2020, 12, 49 2 of 20 arenavirus complex known to cause VHF includes Lassa virus (LASV) and the recently discovered Lujo virus (LUJV) [3,4]. The prototypic arenavirus, lymphocytic choriomeningitis virus (LCMV), which is distributed worldwide, is also of clinical significance [5]. The most prevalent pathogen among the arenaviruses is LASV, the causative agent of Lassa fever (LF), which is a significant cause of morbidity and mortality, with tens of thousands of cases annually and thousands of fatalities in West Africa [6]. The disease is characterized by acute forms associated with fever, myalgia, abdominal pain, nausea, diarrhea, cough, sore throat, and facial edema and evolves toward a shock syndrome in the terminal stage for severe cases. The tendency of LASV to cause outbreaks has steadily risen in Nigeria over the last three years, with laboratory-confirmed cases increasing from 106 in 2016 to 143 in 2017 and reaching 562 by November 2018 [7]. Additionally, there is currently no licensed vaccine or efficient antiviral drug available in the field against this disease, for which the area of endemicity is expanding [8]. The particularity of arenaviruses is the presence of non-pathogenic agents that are not associated with human disease, despite their isolation from the same host species. This is true for Mopeia virus (MOPV), which is closely related to LASV but has never been associated to human infection [9]. MOPV has even been shown to protect non-human primates against a challenge with LASV, showing that both viruses are antigenically related. Comparing LASV and MOPV should therefore allow the identification of immune and viral features involved in LF pathogenesis and, to a lesser extent, other arenavirus-associated VHFs.The bi-s...

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A Map of the Arenavirus Nucleoprotein-Host Protein Interactome Reveals that Junín Virus Selectively Impairs the Antiviral Activity of Double-Stranded RNA-Activated Protein Kinase (PKR)

Cited by 42 publications

References 56 publications

A Proteomics Survey of Junín Virus Interactions with Human Proteins Reveals Host Factors Required for Arenavirus Replication

A Proteomics Survey of Junín Virus Interactions with Human Proteins Reveals Host Factors Required for Arenavirus Replication

Highly Pathogenic New World Arenavirus Infection Activates the Pattern Recognition Receptor Protein Kinase R without Attenuating Virus Replication in Human Cells

E3 Ligase ITCH Interacts with the Z Matrix Protein of Lassa and Mopeia Viruses and Is Required for the Release of Infectious Particles

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