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Cellular and Molecular Life Sciences

, Volume 67, Issue 18, pp 3151–3161 | Cite as

Ultrastructural and quantitative analysis of the lipid droplet clustering induced by hepatitis C virus core protein

  • Marion Depla
  • Rustem Uzbekov
  • Christophe Hourioux
  • Emmanuelle Blanchard
  • Amélie Le Gouge
  • Ludovic Gillet
  • Philippe RoingeardEmail author
Research Article

Abstract

Hepatitis C virus (HCV) release is linked to the formation of lipid droplet (LD) clusters in the perinuclear area of infected cells, induced by the core protein. We used electron microscopy (EM) to monitor and compare the number and size of LD in cells producing the mature and immature forms of the HCV core protein, and 3D EM to reconstruct whole cells producing the mature core protein. Only the mature protein coated the LD and induced their clustering and emergence from endoplasmic reticulum membranes enriched in this protein. We found no particular association between LD clusters and the centrosome in reconstructed cells. The LD clustering induced by the mature core protein was associated with an increase in LD synthesis potentially due, at least in part, to the ability of this protein to coat the LD. These observations provide useful information for further studies of the mechanisms involved in HCV-induced steatosis.

Keywords

HCV Lipid droplet Steatosis Electron microscopy 3D reconstruction 

Abbreviations

HCV

Hepatitis C virus

ER

Endoplasmic reticulum

SPP

Signal peptide peptidase

SFV

Semliki forest virus

EM

Electron microscopy

β-Gal

β-Galactosidase

WT

Wild-type

Notes

Acknowledgment

This work was supported by a grant INSERM-DHOS « Virosteatose ». M.D. was supported by a fellowship from the INSERM and Région Centre. We thank Dr Mario Mondelli (Istituto di Clinica delle Malattie Infettive, Pavia, Italy) for providing us with the monoclonal B12F8 anti-HCV core reagent. We thank Sylvie Trassard and Fabienne Arcanger for technical assistance. We thank Bruno Giraudeau, Eric Piver and Pierre Besson for helpful discussions and feedback on this work. Our data were obtained with the assistance of the RIO Electron Microscopy Facility of François Rabelais University.

Supplementary material

Supplementary material 1 (MPEG 6214 kb)

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Copyright information

© Springer Basel AG 2010

Authors and Affiliations

  • Marion Depla
    • 1
  • Rustem Uzbekov
    • 1
  • Christophe Hourioux
    • 1
  • Emmanuelle Blanchard
    • 1
  • Amélie Le Gouge
    • 2
  • Ludovic Gillet
    • 3
  • Philippe Roingeard
    • 1
    Email author
  1. 1.INSERM U966, Faculté de MédecineUniversité François Rabelais, CHRU de ToursTours CedexFrance
  2. 2.INSERM CIC 0202Université François Rabelais, CHRU de ToursTours CedexFrance
  3. 3.INSERM U921Université François Rabelais, CHRU de ToursTours CedexFrance

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