Inhibition of Cytosolic Phospholipase A
2
α Impairs an Early Step of Coronavirus Replication in Cell Culture
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Published:2018-02-15
Issue:4
Volume:92
Page:
-
ISSN:0022-538X
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Container-title:Journal of Virology
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language:en
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Short-container-title:J Virol
Author:
Müller Christin1, Hardt Martin2, Schwudke Dominik3, Neuman Benjamin W.4, Pleschka Stephan1, Ziebuhr John1
Affiliation:
1. Institute of Medical Virology, Justus Liebig University Giessen, Giessen, Germany 2. Imaging Unit, Biomedical Research Center, Justus Liebig University Giessen, Giessen, Germany 3. Division of Bioanalytical Chemistry, Priority Area Infection, Research Center Borstel, Leibniz Center for Medicine and Bioscience, Borstel, Germany 4. Texas A&M University, Texarkana, Texas, USA
Abstract
ABSTRACT
Coronavirus replication is associated with intracellular membrane rearrangements in infected cells, resulting in the formation of double-membrane vesicles (DMVs) and other membranous structures that are referred to as replicative organelles (ROs). The latter provide a structural scaffold for viral replication/transcription complexes (RTCs) and help to sequester RTC components from recognition by cellular factors involved in antiviral host responses. There is increasing evidence that plus-strand RNA (+RNA) virus replication, including RO formation and virion morphogenesis, affects cellular lipid metabolism and critically depends on enzymes involved in lipid synthesis and processing. Here, we investigated the role of cytosolic phospholipase A
2
α (cPLA
2
α) in coronavirus replication using a low-molecular-weight nonpeptidic inhibitor, pyrrolidine-2 (Py-2). The inhibition of cPLA
2
α activity, which produces lysophospholipids (LPLs) by cleaving at the
sn
-2 position of phospholipids, had profound effects on viral RNA and protein accumulation in human coronavirus 229E-infected Huh-7 cells. Transmission electron microscopy revealed that DMV formation in infected cells was significantly reduced in the presence of the inhibitor. Furthermore, we found that (i) viral RTCs colocalized with LPL-containing membranes, (ii) cellular LPL concentrations were increased in coronavirus-infected cells, and (iii) this increase was diminished in the presence of the cPLA
2
α inhibitor Py-2. Py-2 also displayed antiviral activities against other viruses representing the
Coronaviridae
and
Togaviridae
families, while members of the
Picornaviridae
were not affected. Taken together, the study provides evidence that cPLA
2
α activity is critically involved in the replication of various +RNA virus families and may thus represent a candidate target for broad-spectrum antiviral drug development.
IMPORTANCE
Examples of highly conserved RNA virus proteins that qualify as drug targets for broad-spectrum antivirals remain scarce, resulting in increased efforts to identify and specifically inhibit cellular functions that are essential for the replication of RNA viruses belonging to different genera and families. The present study supports and extends previous conclusions that enzymes involved in cellular lipid metabolism may be tractable targets for broad-spectrum antivirals. We obtained evidence to show that a cellular phospholipase, cPLA2α, which releases fatty acid from the
sn
-2 position of membrane-associated glycerophospholipids, is critically involved in coronavirus replication, most likely by producing lysophospholipids that are required to form the specialized membrane compartments in which viral RNA synthesis takes place. The importance of this enzyme in coronavirus replication and DMV formation is supported by several lines of evidence, including confocal and electron microscopy, viral replication, and lipidomics studies of coronavirus-infected cells treated with a highly specific cPLA
2
α inhibitor.
Funder
German Center for Infection Research Deutsche Forschungsgemeinschaft
Publisher
American Society for Microbiology
Subject
Virology,Insect Science,Immunology,Microbiology
Reference92 articles.
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