Low pH-dependent hepatitis C virus membrane fusion depends on E2 integrity, target lipid composition, and density of virus particles.

2.50
Hdl Handle:
http://hdl.handle.net/10033/78494
Title:
Low pH-dependent hepatitis C virus membrane fusion depends on E2 integrity, target lipid composition, and density of virus particles.
Authors:
Haid, Sibylle; Pietschmann, Thomas; Pécheur, Eve-Isabelle
Abstract:
Hepatitis C virus (HCV) is an enveloped, positive strand RNA virus of about 9.6 kb. Like all enveloped viruses, the HCV membrane fuses with the host cell membrane during the entry process and thereby releases the genome into the cytoplasm, initiating the viral replication cycle. To investigate the features of HCV membrane fusion, we developed an in vitro fusion assay using cell culture-produced HCV and fluorescently labeled liposomes. With this model we could show that HCV-mediated fusion can be triggered in a receptor-independent but pH-dependent manner and that fusion of the HCV particles with liposomes is dependent on the viral dose and on the lipid composition of the target membranes. In addition CBH-5, an HCV E2-specific antibody, inhibited fusion in a dose-dependent manner. Interestingly, point mutations in E2, known to abrogate HCV glycoprotein-mediated fusion in a cell-based assay, altered or even abolished fusion in the liposome-based assay. When assaying the fusion properties of HCV particles with different buoyant density, we noted higher fusogenicity of particles with lower density. This could be attributable to inherently different properties of low density particles, to association of these particles with factors stimulating fusion, or to co-flotation of factors enhancing fusion activity in trans. Taken together, these data show the important role of lipids of both the viral and target membranes in HCV-mediated fusion, point to a crucial role played by the E2 glycoprotein in the process of HCV fusion, and reveal an important behavior of HCV of different densities with regard to fusion.
Affiliation:
Department for Experimental Virology, TWINCORE, Centre for Experimental and Clinical Infection Research, a Joint Venture of Hannover Medical School and the Helmholtz-Centre for Infection Research, Hannover 30625, Germany.
Citation:
Low pH-dependent hepatitis C virus membrane fusion depends on E2 integrity, target lipid composition, and density of virus particles. 2009, 284 (26):17657-67 J. Biol. Chem.
Journal:
The Journal of biological chemistry
Issue Date:
26-Jun-2009
URI:
http://hdl.handle.net/10033/78494
DOI:
10.1074/jbc.M109.014647
PubMed ID:
19411248
Type:
Article
Language:
en
ISSN:
0021-9258
Appears in Collections:
publications of the department experimental Virology([TC]EVIR)

Full metadata record

DC FieldValue Language
dc.contributor.authorHaid, Sibylle-
dc.contributor.authorPietschmann, Thomas-
dc.contributor.authorPécheur, Eve-Isabelle-
dc.date.accessioned2009-08-25T13:22:55Z-
dc.date.available2009-08-25T13:22:55Z-
dc.date.issued2009-06-26-
dc.identifier.citationLow pH-dependent hepatitis C virus membrane fusion depends on E2 integrity, target lipid composition, and density of virus particles. 2009, 284 (26):17657-67 J. Biol. Chem.en
dc.identifier.issn0021-9258-
dc.identifier.pmid19411248-
dc.identifier.doi10.1074/jbc.M109.014647-
dc.identifier.urihttp://hdl.handle.net/10033/78494-
dc.description.abstractHepatitis C virus (HCV) is an enveloped, positive strand RNA virus of about 9.6 kb. Like all enveloped viruses, the HCV membrane fuses with the host cell membrane during the entry process and thereby releases the genome into the cytoplasm, initiating the viral replication cycle. To investigate the features of HCV membrane fusion, we developed an in vitro fusion assay using cell culture-produced HCV and fluorescently labeled liposomes. With this model we could show that HCV-mediated fusion can be triggered in a receptor-independent but pH-dependent manner and that fusion of the HCV particles with liposomes is dependent on the viral dose and on the lipid composition of the target membranes. In addition CBH-5, an HCV E2-specific antibody, inhibited fusion in a dose-dependent manner. Interestingly, point mutations in E2, known to abrogate HCV glycoprotein-mediated fusion in a cell-based assay, altered or even abolished fusion in the liposome-based assay. When assaying the fusion properties of HCV particles with different buoyant density, we noted higher fusogenicity of particles with lower density. This could be attributable to inherently different properties of low density particles, to association of these particles with factors stimulating fusion, or to co-flotation of factors enhancing fusion activity in trans. Taken together, these data show the important role of lipids of both the viral and target membranes in HCV-mediated fusion, point to a crucial role played by the E2 glycoprotein in the process of HCV fusion, and reveal an important behavior of HCV of different densities with regard to fusion.en
dc.language.isoenen
dc.subject.meshCarcinoma, Hepatocellularen
dc.subject.meshCholesterolen
dc.subject.meshElectroporationen
dc.subject.meshHepacivirusen
dc.subject.meshHepatitis Cen
dc.subject.meshHepatitis C Antibodiesen
dc.subject.meshHumansen
dc.subject.meshHydrogen-Ion Concentrationen
dc.subject.meshImmunoenzyme Techniquesen
dc.subject.meshImmunoprecipitationen
dc.subject.meshIndolesen
dc.subject.meshLiposomesen
dc.subject.meshLiver Neoplasmsen
dc.subject.meshLuciferasesen
dc.subject.meshRNA, Viralen
dc.subject.meshSphingomyelinsen
dc.subject.meshTranscription, Geneticen
dc.subject.meshTumor Cells, Cultureden
dc.subject.meshViral Envelope Proteinsen
dc.subject.meshVirionen
dc.subject.meshVirus Internalizationen
dc.titleLow pH-dependent hepatitis C virus membrane fusion depends on E2 integrity, target lipid composition, and density of virus particles.en
dc.typeArticleen
dc.contributor.departmentDepartment for Experimental Virology, TWINCORE, Centre for Experimental and Clinical Infection Research, a Joint Venture of Hannover Medical School and the Helmholtz-Centre for Infection Research, Hannover 30625, Germany.en
dc.identifier.journalThe Journal of biological chemistryen

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