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Sequential biogenesis of host cell membrane rearrangements induced by hepatitis C virus infection

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Abstract

Like most positive-strand RNA viruses, hepatitis C virus (HCV) forms a membrane-associated replication complex consisting of replicating RNA, viral and host proteins anchored to altered cell membranes. We used a combination of qualitative and quantitative electron microscopy (EM), immuno-EM, and the 3D reconstruction of serial EM sections to analyze the host cell membrane alterations induced by HCV. Three different types of membrane alteration were observed: vesicles in clusters (ViCs), contiguous vesicles (CVs), and double-membrane vesicles (DMVs). The main ultrastructural change observed early in infection was the formation of a network of CVs surrounding the lipid droplets. Later stages in the infectious cycle were characterized by a large increase in the number of DMVs, which may be derived from the CVs. These DMVs are thought to constitute the membranous structures harboring the viral replication complexes in which viral replication is firmly and permanently established and to protect the virus against double-stranded RNA-triggered host antiviral responses.

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Abbreviations

BSA:

Bovine serum albumin

CM:

Convoluted membranes

CV:

Contiguous vesicles

DENV:

Dengue virus

DMEM:

Dulbecco’s modified Eagle medium

DMV:

Double-membrane vesicle

EM:

Electron microscopy

FFU:

Focus forming unit

HCV:

Hepatitis C virus

KUNV:

Kunjin virus

LD:

Lipid droplet

MAbs:

Mouse monoclonal antibody

MMV:

Multimembrane vesicles

MVB:

Multivesicular bodies

ViC:

Vesicles in cluster

VP:

Vesicle packets

WNV:

West Nile virus

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Acknowledgments

This work was supported by a grant from the ANRS (Agence Nationale pour la Recherche sur le Sida et les hépatites virales), France. P.F. was supported by a PhD fellowship from the ANRS. We thank Dr Charles Rice and Dr Takaji Wakita for providing us with the Huh7.5 cell line and the JFH-1 virus strain, respectively. We thank Fabienne Arcanger and Juliette Rousseau for technical assistance with EM sections. We thank Catherine Gaudy and Alain Goudeau for HCV RNA quantifications and Christophe Hourioux and Jean-Christophe Meunier for helpful discussions on this work. Our data were obtained with the assistance of the RIO Electron Microscopy Facility of François Rabelais University.

Author information

Correspondence to Philippe Roingeard.

Additional information

E. Blanchard and P. Roingeard are co-last authors.

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Supplementary material 3 (MPG 9697 kb)

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Ferraris, P., Beaumont, E., Uzbekov, R. et al. Sequential biogenesis of host cell membrane rearrangements induced by hepatitis C virus infection. Cell. Mol. Life Sci. 70, 1297–1306 (2013). https://doi.org/10.1007/s00018-012-1213-0

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Keywords

  • HCV
  • Host cell membranes
  • Membranous web
  • Electron microscopy
  • 3D reconstruction