Characterization of Junín arenavirus cell entry

Martínez, María del Carmen Jiménez; Cordo, Sandra M.; Candurra, Nélida A.

doi:10.1099/vir.0.82808-0

Cited by 94 publications

(91 citation statements)

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“…Upon receptor binding, arenaviruses undergo endocytosis and are delivered to acidified endosomes, where GP-mediated pH-dependent fusion between the viral and the cellular membrane occurs (4,8). Consistent with the use of TfR1 as a primary receptor, JUNV enters cells via clathrin-mediated endocytosis (30,46). In contrast, no particular endocytotic pathway has been associated with ␣-DG.…”

mentioning

confidence: 88%

Different Mechanisms of Cell Entry by Human-Pathogenic Old World and New World Arenaviruses

Rojek

Sánchez

Nguyen

et al. 2008

J Virol

133

142

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The Old World arenavirus Lassa virus (LASV) is the causative agent of severe viral hemorrhagic fever (VHF) in humans and is the most prevalent human pathogen among arenaviruses. The present study investigated the largely unknown mechanisms of cell entry of LASV, a process know to be mediated solely by the virus envelope glycoprotein (GP). To circumvent biosafety restrictions associated with the use of live LASV, we used reverse genetics to generate a recombinant variant of the prototypic arenavirus lymphocytic choriomeningitis virus (LCMV) expressing the LASV GP (rLCMV-LASVGP). The rescued rLCMV-LASVGP grew to titers comparable to that of LCMV and showed the receptor binding characteristics of LASV. We used rLCMV-LASVGP to characterize the cellular mechanisms of LASV entry in the context of a productive arenavirus infection. The kinetics of pH-dependent membrane fusion of rLCMV-LASVGP resembled those of the human-pathogenic New World arenavirus Junin virus (JUNV) and other enveloped viruses that use clathrin-mediated endocytosis for entry. However, rLCMV-LASVGP entered cells predominantly via a clathrin-, caveolin-, and dynamin-independent endocytotic pathway similar to the one recently described for LCMV. Productive infection of rLCMV-LASVGP was only mildly affected by a dominant negative mutant of Rab5 and was independent of Rab7, suggesting an unusual mechanism of delivery to endosomes. In addition, rLCMV-LASVGP infection was independent of actin but required intact microtubules. Our data indicate that LASV enters cells via a pathway distinct from the one used by human-pathogenic New World arenaviruses.

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mentioning

confidence: 88%

Different Mechanisms of Cell Entry by Human-Pathogenic Old World and New World Arenaviruses

Rojek

Sánchez

Nguyen

et al. 2008

J Virol

133

142

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“…Fusion allows the release of the ribonucleoproteins (RNPs), composed of the viral genomic RNA protected by NP proteins, into the cytosol of the infected cell. Available evidence supports the possibility that the internalization of JUNV depends on clathrin-mediated endocytosis and requires subsequent endosomal acidification for membrane fusion and penetration (9)(10)(11). The early and late endosomal compartments, as identified by the small GTPases Rab5 and Rab7, respectively, have also been proposed to be important for efficient JUNV infection (12).…”

Section: T He Junin Virus (Junv) Is An Important Member Of the Newmentioning

confidence: 99%

Identification and Characterization of a Novel Broad-Spectrum Virus Entry Inhibitor

Chou

Cuevas

Carocci

et al. 2016

J Virol

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Virus entry into cells is a multistep process that often requires the subversion of subcellular machineries. A more complete understanding of these steps is necessary to develop new antiviral strategies. While studying the potential role of the actin network and one of its master regulators, the small GTPase Cdc42, during Junin virus (JUNV) entry, we serendipitously uncovered the small molecule ZCL278, reported to inhibit Cdc42 function as an entry inhibitor for JUNV and for vesicular stomatitis virus, lymphocytic choriomeningitis virus, and dengue virus but not for the nonenveloped poliovirus. Although ZCL278 did not interfere with JUNV attachment to the cell surface or virus particle internalization into host cells, it prevented the release of JUNV ribonucleoprotein cores into the cytosol and decreased pH-mediated viral fusion with host membranes. We also identified SVG-A astroglial cell-derived cells to be highly permissive for JUNV infection and generated new cell lines expressing fluorescently tagged Rab5c or Rab7a or lacking Cdc42 using clustered regularly interspaced short palindromic repeat (CRISPR)-caspase 9 (Cas9) gene-editing strategies. Aided by these tools, we uncovered that perturbations in the actin cytoskeleton or Cdc42 activity minimally affect JUNV entry, suggesting that the inhibitory effect of ZCL278 is not mediated by ZCL278 interfering with the activity of Cdc42. Instead, ZCL278 appears to redistribute viral particles from endosomal to lysosomal compartments. ZCL278 also inhibited JUNV replication in a mouse model, and no toxicity was detected. Together, our data suggest the unexpected antiviral activity of ZCL278 and highlight its potential for use in the development of valuable new tools to study the intracellular trafficking of pathogens. IMPORTANCEThe Junin virus is responsible for outbreaks of Argentine hemorrhagic fever in South America, where 5 million people are at risk. Limited options are currently available to treat infections by Junin virus or other viruses of the Arenaviridae, making the identification of additional tools, including small-molecule inhibitors, of great importance. How Junin virus enters cells is not yet fully understood. Here we describe new cell culture models in which the cells are susceptible to Junin virus infection and to which we applied CRISPR-Cas9 genome engineering strategies to help characterize early steps during virus entry. We also uncovered ZCL278 to be a new antiviral small molecule that potently inhibits the cellular entry of the Junin virus and other enveloped viruses. Moreover, we show that ZCL278 also functions in vivo, thereby preventing Junin virus replication in a mouse model, opening the possibility for the discovery of ZCL278 derivatives of therapeutic potential. T he Junin virus (JUNV) is an important member of the NewWorld arenavirus family (clade B) and is responsible for outbreaks of Argentine hemorrhagic fever in South America, where 5 million people are considered to be at risk (1). The reservoir of JUNV is the rodent Calomy...

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“…Next, we investigated drug effects on endocytotic pathways implicated in arenavirus entry, clathrin-mediated endocytosis proposed for JUNV (28,29) and cholesterol-dependent endocytotic pathways implicated in LASV entry (28). Using well established cell biological assays for clathrin-mediated and cholesterol-dependent lipid raft-mediated endocytosis utilizing transferrin and cholera toxin B as markers, we found no inhibitory effect of our compounds at 20-fold IC 50 (supplemental Fig.…”

Section: High Throughput Screening Of Combinatorial Chemicalmentioning

confidence: 99%