Functional analysis of hepatitis C virus envelope proteins, using a cell-cell fusion assay

Mariko Kobayashi, Michael C. Bennett, Theodore Bercot, Ila R. Singh

Research output: Contribution to journalArticlepeer-review

43 Scopus citations

Abstract

Hepatitis C virus (HCV) envelope proteins mediate the entry of virus into cells by binding to cellular receptors, resulting in fusion of the viral membrane with the host cell membrane and permitting the viral genome to enter the cytoplasm. We report the development of a robust and reproducible cell-cell fusion assay using envelope proteins from commonly occurring genotypes of HCV. The assay scored HCV envelope protein-mediated fusion by the production of fluorescent green syncytia and allowed us to elucidate many aspects of HCV fusion, including the pH of fusion, cell types that permit viral entry, and the conformation of envelope proteins essential for fusion. We found that fusion could be specifically inhibited by anti-HCV antibodies and by at least one peptide. We also generated a number of insertional mutations in the envelope proteins and tested nine of these using the fusion assay. We demonstrate that this fusion assay is a powerful tool for understanding the mechanism of HCV-mediated fusion, elucidating mutant function, and testing antiviral agents.

Original languageEnglish
Pages (from-to)1817-1825
Number of pages9
JournalJournal of Virology
Volume80
Issue number4
DOIs
StatePublished - Feb 2006
Externally publishedYes

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