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The lipid droplet is an important organelle for hepatitis C virus production

Nature Cell Biology volume 9, pages 1089–1097 (2007)Cite this article

An Erratum to this article was published on 01 October 2007

Abstract

The lipid droplet (LD) is an organelle that is used for the storage of neutral lipids. It dynamically moves through the cytoplasm, interacting with other organelles, including the endoplasmic reticulum (ER)1,2,3. These interactions are thought to facilitate the transport of lipids and proteins to other organelles. The hepatitis C virus (HCV) is a causative agent of chronic liver diseases4. HCV capsid protein (Core) associates with the LD5, envelope proteins E1 and E2 reside in the ER lumen6, and the viral replicase is assumed to localize on ER-derived membranes. How and where HCV particles are assembled, however, is poorly understood. Here, we show that the LD is involved in the production of infectious virus particles. We demonstrate that Core recruits nonstructural (NS) proteins and replication complexes to LD-associated membranes, and that this recruitment is critical for producing infectious viruses. Furthermore, virus particles were observed in close proximity to LDs, indicating that some steps of virus assembly take place around LDs. This study reveals a novel function of LDs in the assembly of infectious HCV and provides a new perspective on how viruses usurp cellular functions.

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Figure 1: Core recruits NS proteins to LDs.
Figure 2: Core-dependent recruitment of active HCV replication complexes to the LD.
Figure 3: Spatial distribution of Core and NS5A relative to the LD.
Figure 4: LD associations of Core and NS proteins are necessary for the production of infectious HCV particles.
Figure 5: Virus-assembly takes place around the LDs.

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References

  1. Martin, S. & Parton, R. G. Lipid droplets: a unified view of a dynamic organelle. Nature Rev. Mol. Cell Biol. 7, 373–378 (2006).

    Article  CAS  Google Scholar 

  2. Blanchette-Mackie, E. J. et al. Perilipin is located on the surface layer of intracellular lipid droplets in adipocytes. J. Lipid Res. 36, 1211–1226 (1995).

    CAS  PubMed  Google Scholar 

  3. Vock, R. et al. Design of the oxygen and substrate pathways. VI. structural basis of intracellular substrate supply to mitochondria in muscle cells. J. Exp. Biol. 199, 1689–1697 (1996).

    CAS  PubMed  Google Scholar 

  4. Liang, T. J. et al. Viral pathogenesis of hepatocellular carcinoma in the United States. Hepatology 18, 1326–1333 (1993).

    CAS  PubMed  Google Scholar 

  5. Moradpour, D., Englert, C., Wakita, T. & Wands, J. R. Characterization of cell lines allowing tightly regulated expression of hepatitis C virus core protein. Virology 222, 51–63 (1996).

    Article  CAS  Google Scholar 

  6. Deleersnyder, V. et al. Formation of native hepatitis C virus glycoprotein complexes. J. Virol. 71, 697–704 (1997).

    CAS  PubMed  PubMed Central  Google Scholar 

  7. Kato, N. et al. Molecular cloning of the human hepatitis C virus genome from Japanese patients with non-A, non-B hepatitis. Proc. Natl Acad. Sci. USA 87, 9524–9528 (1990).

    Article  CAS  Google Scholar 

  8. Hijikata, M. & Shimotohno, K. [Mechanisms of hepatitis C viral polyprotein processing]. Uirusu 43, 293–298 (1993).

    Article  CAS  Google Scholar 

  9. Dubuisson, J., Penin, F. & Moradpour, D. Interaction of hepatitis C virus proteins with host cell membranes and lipids. Trends Cell Biol. 12, 517–523 (2002).

    Article  CAS  Google Scholar 

  10. Wakita, T. et al. Production of infectious hepatitis C virus in tissue culture from a cloned viral genome. Nature Med. 11, 791–796 (2005).

    Article  CAS  Google Scholar 

  11. Lindenbach, B. D. et al. Complete replication of hepatitis C virus in cell culture. Science 309, 623–626 (2005).

    Article  CAS  Google Scholar 

  12. Zhong, J. et al. Robust hepatitis C virus infection in vitro. Proc. Natl Acad. Sci. USA 102, 9294–9299 (2005).

    Article  CAS  Google Scholar 

  13. Pietschmann, T. et al. Construction and characterization of infectious intragenotypic and intergenotypic hepatitis C virus chimeras. Proc. Natl Acad. Sci. USA 103, 7408–7413 (2006).

    Article  CAS  Google Scholar 

  14. Moriya, K. et al. Hepatitis C virus core protein induces hepatic steatosis in transgenic mice. J. Gen. Virol. 78, 1527–1531 (1997).

    Article  CAS  Google Scholar 

  15. Hope, R. G., Murphy, D. J. & McLauchlan, J. The domains required to direct core proteins of hepatitis C virus and GB virus-B to lipid droplets share common features with plant oleosin proteins. J. Biol. Chem. 277, 4261–4270 (2002).

    Article  CAS  Google Scholar 

  16. Egger, D. et al. Expression of hepatitis C virus proteins induces distinct membrane alterations including a candidate viral replication complex. J. Virol. 76, 5974–5984 (2002).

    Article  CAS  Google Scholar 

  17. Miyanari, Y. et al. Hepatitis C virus non-structural proteins in the probable membranous compartment function in viral genome replication. J. Biol. Chem. 278, 50301–50308 (2003).

    Article  CAS  Google Scholar 

  18. Quinkert, D., Bartenschlager, R. & Lohmann, V. Quantitative analysis of the hepatitis C virus replication complex. J. Virol. 79, 13594–13605 (2005).

    Article  CAS  Google Scholar 

  19. Tauchi-Sato, K., Ozeki, S., Houjou, T., Taguchi, R. & Fujimoto, T. The surface of lipid droplets is a phospholipid monolayer with a unique fatty acid composition. J. Biol. Chem. 277, 44507–44512 (2002).

    Article  CAS  Google Scholar 

  20. Londos, C., Brasaemle, D. L., Schultz, C. J., Segrest, J. P. & Kimmel, A. R. Perilipins, ADRP, and other proteins that associate with intracellular neutral lipid droplets in animal cells. Semin. Cell Dev. Biol. 10, 51–58 (1999).

    Article  CAS  Google Scholar 

  21. Blight, K. J., McKeating, J. A. & Rice, C. M. Highly permissive cell lines for subgenomic and genomic hepatitis C virus RNA replication. J. Virol. 76, 13001–13014 (2002).

    Article  CAS  Google Scholar 

  22. Klein, K. C., Dellos, S. R. & Lingappa, J. R. Identification of residues in the hepatitis C virus core protein that are critical for capsid assembly in a cell-free system. J. Virol. 79, 6814–6826 (2005).

    Article  CAS  Google Scholar 

  23. Owsianka, A. et al. Monoclonal antibody AP33 defines a broadly neutralizing epitope on the hepatitis C virus E2 envelope glycoprotein. J Virol 79, 11095–11104 (2005).

    Article  CAS  Google Scholar 

  24. Ishii, N. et al. Diverse effects of cyclosporine on hepatitis C virus strain replication. J. Virol. 80, 4510–4520 (2006).

    Article  CAS  Google Scholar 

  25. Lohmann, V., Korner, F., Herian, U. & Bartenschlager, R. Biochemical properties of hepatitis C virus NS5B RNA-dependent RNA polymerase and identification of amino acid sequence motifs essential for enzymatic activity. J Virol 71, 8416–8428 (1997).

    CAS  PubMed  PubMed Central  Google Scholar 

  26. Ohsaki, Y., Maeda, T. & Fujimoto, T. Fixation and permeabilization protocol is critical for the immunolabeling of lipid droplet proteins. Histochem. Cell Biol. 124, 445–452 (2005).

    Article  CAS  Google Scholar 

Download references

Acknowledgements

We thank T. Fujimoto and Y. Ohsaki at Nagoya University for helpful discussions and technical assistance. Y.M. is a recipient of a JSPS fellowship. K.S. is supported by Grants-in-Aid for cancer research and for the second-term comprehensive 10-year strategy for cancer control from the Ministry of Health, Labour and Welfare, as well as by a Grant-in-Aid for Scientific Research on Priority Areas “Integrative Research Toward the Conquest of Cancer” from the Ministry of Education, Culture, Sports, Science and Technology of Japan. T.W. is also supported, in part, by a Grant-in-Aid for Scientific Research from the Japan Society for the Promotion of Science; and by the Research on Health Sciences Focusing on Drug Innovation from the Japan Health Sciences Foundation. R.B. is supported by the Sonderforschungsbereich 638 (Teilprojekt A5) and the Deutsche Forschungsgemeinschaft (BA1505/2-1). M.Z. and R.B. thank the Nikon Imaging Center at the University of Heidelberg for providing access to their confocal fluorescence microscopes and Ulrike Engel for the excellent support.

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Authors and Affiliations

  1. Department of Viral Oncology, Institute for Virus Research, Kyoto University, Kyoto, 606-8507, Japan

    Yusuke Miyanari, Koichi Watashi, Takayuki Hishiki, Makoto Hijikata & Kunitada Shimotohno

  2. Graduate School of Biostudies, Kyoto University, Kyoto, 606-8507, Japan

    Yusuke Miyanari, Koichi Watashi, Takayuki Hishiki, Makoto Hijikata & Kunitada Shimotohno

  3. Department of Anatomy, Fujita Health University School of Medicine, Toyoake, 470-1192, Japan

    Kimie Atsuzawa & Nobuteru Usuda

  4. Department of Molecular Virology, University of Heidelberg, 69120, Heidelberg, Germany

    Margarita Zayas & Ralf Bartenschlager

  5. Department of Virology II, National Institute of Infectious Diseases, Tokyo, 162-8640, Japan

    Takaji Wakita

  6. Correspondence should be addressed to K.S. (e-mail: ),

    Kunitada Shimotohno

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  1. Yusuke Miyanari
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Contributions

Y.M. and K.S. planned experiments and analyses. Y.M. was responsible for experiments for Figs 1, 2, 3a–c, 4a–e and 5b. K.A., N.U., electron microscopy; T.H., Fig. 1e; M.Z., R.B., Fig. S2e; and K.S. and K.W., Fig. 4f–g. T.W. provided JFH1 strain. Y.M. and K.S. wrote the manuscript. All authors discussed the results and commented on the manuscript.

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Supplementary Figures 1, 2, 3, 4, 5 and 6, Supplementary Table, Supplementary Materials and Methods (PDF 2664 kb)

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Miyanari, Y., Atsuzawa, K., Usuda, N. et al. The lipid droplet is an important organelle for hepatitis C virus production. Nat Cell Biol 9, 1089–1097 (2007). https://doi.org/10.1038/ncb1631

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