1887

Abstract

Recently, the characterization of a cell culture system allowing the amplification of an authentic virus, named hepatitis C virus cell culture (HCVcc), has been reported by several groups. To obtain higher HCV particle productions, we investigated the potential effect of some amino acid changes on the infectivity of the JFH-1 isolate. As a first approach, successive infections of naïve Huh-7 cells were performed until high viral titres were obtained, and mutations that appeared during this selection were identified by sequencing. Only one major modification, N534K, located in the E2 glycoprotein sequence was found. Interestingly, this mutation prevented core glycosylation of E2 site 6. In addition, JFH-1 generated with this modification facilitated the infection of Huh-7 cells. In a second approach to identify mutations favouring HCVcc infectivity, we exploited the observation that a chimeric virus containing the genotype 1a core protein in the context of JFH-1 background was more infectious than wild-type JFH-1 isolate. Sequence alignment between JFH-1 and our chimera, led us to identify two major positions, 172 and 173, which were not occupied by similar amino acids in these two viruses. Importantly, higher viral titres were obtained by introducing these residues in the context of wild-type JFH-1. Altogether, our data indicate that a more robust production of HCVcc particles can be obtained by introducing a few specific mutations in JFH-1 structural proteins.

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2007-09-01
2024-03-29
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References

  1. Bartosch B., Dubuisson J., Cosset F. L. 2003; Infectious hepatitis C virus pseudo-particles containing functional E1–E2 envelope protein complexes. J Exp Med 197:633–642 [CrossRef]
    [Google Scholar]
  2. Blight K. J., Kolykhalov A. A., Rice C. M. 2000; Efficient initiation of HCV RNA replication in cell culture. Science 290:1972–1974 [CrossRef]
    [Google Scholar]
  3. Castelain S., Descamps V., Thibault V., Francois C., Bonte D., Morel V., Izopet J., Capron D., Zawadzki P. other authors 2004; TaqMan amplification system with an internal positive control for HCV RNA quantitation. J Clin Virol 31:227–234 [CrossRef]
    [Google Scholar]
  4. Drummer H. E., Boo I., Maerz A. L., Poumbourios P. 2006; A conserved Gly436-Trp-Leu-Ala-Gly-Leu-Phe-Tyr motif in hepatitis C virus glycoprotein E2 is a determinant of CD81 binding and viral entry. J Virol 80:7844–7853 [CrossRef]
    [Google Scholar]
  5. Flint M., Maidens C., Loomis-Price L. D., Shotton C., Dubuisson J., Monk P., Higginbottom A., Levy S., McKeating J. A. 1999; Characterization of hepatitis C virus E2 glycoprotein interaction with a putative cellular receptor, CD81. J Virol 73:6235–6244
    [Google Scholar]
  6. Goffard A., Callens N., Bartosch B., Wychowski C., Cosset F. L., Montpellier C., Dubuisson J. 2005; Role of N-linked glycans in the functions of hepatitis C virus envelope glycoproteins. J Virol 79:8400–8409 [CrossRef]
    [Google Scholar]
  7. Grakoui A., Wychowski C., Lin C., Feinstone S. M., Rice C. M. 1993; Expression and identification of hepatitis C virus polyprotein cleavage products. J Virol 67:1385–1395
    [Google Scholar]
  8. Griffin S. D., Beales L. P., Clarke D. S., Worsfold O., Evans S. D., Jaeger J., Harris M. P., Rowlands D. J. 2003; The p7 protein of hepatitis C virus forms an ion channel that is blocked by the antiviral drug, Amantadine. FEBS Lett 535:34–38 [CrossRef]
    [Google Scholar]
  9. Hijikata M., Kato N., Ootsuyama Y., Nakagawa M., Shimotohno K. 1991; Gene mapping of the putative structural region of the hepatitis C virus genome by in vitro processing analysis. Proc Natl Acad Sci U S A 88:5547–5551 [CrossRef]
    [Google Scholar]
  10. Houghton M. 1996; Hepatitis C viruses. In Fields Virology . , 3rd edn. pp 1035–1058 Edited by Fields B. N., Knipe D. M., Howley P. M. Philadelphia, PA: Lippincott-Raven;
  11. Hsu M., Zhang J., Flint M., Logvinoff C., Cheng-Mayer C., Rice C. M., McKeating J. A. 2003; Hepatitis C virus glycoproteins mediate pH-dependent cell entry of pseudotyped retroviral particles. Proc Natl Acad Sci U S A 100:7271–7276 [CrossRef]
    [Google Scholar]
  12. Hussy P., Langen H., Mous J., Jacobsen H. 1996; Hepatitis C virus core protein: carboxy-terminal boundaries of two processed species suggest cleavage by a signal peptide peptidase. Virology 224:93–104 [CrossRef]
    [Google Scholar]
  13. Kato T., Furusaka A., Miyamoto M., Date T., Yasui K., Hiramoto J., Nagayama K., Tanaka T., Wakita T. 2001; Sequence analysis of hepatitis C virus isolated from a fulminant hepatitis patient. J Med Virol 64:334–339 [CrossRef]
    [Google Scholar]
  14. Kato T., Date T., Miyamoto M., Furusaka A., Tokushige K., Mizokami M., Wakita T. 2003a; Efficient replication of the genotype 2a hepatitis C virus subgenomic replicon. Gastroenterology 125:1808–1817 [CrossRef]
    [Google Scholar]
  15. Kato T., Miyamoto M., Furusaka A., Date T., Yasui K., Kato J., Matsushima S., Komatsu T., Wakita T. 2003b; Processing of hepatitis C virus core protein is regulated by its C-terminal sequence. J Med Virol 69:357–366 [CrossRef]
    [Google Scholar]
  16. Kolykhalov A. A., Agapov E. V., Blight K. J., Mihalik K., Feinstone S. M., Rice C. M. 1997; Transmission of hepatitis C by intrahepatic inoculation with transcribed RNA. Science 277:570–574 [CrossRef]
    [Google Scholar]
  17. Lavillette D., Tarr A. W., Voisset C., Donot P., Bartosch B., Bain C., Patel A. H., Dubuisson J., Ball J. K. other authors 2005; Characterization of host-range and cell entry properties of the major genotypes and subtypes of hepatitis C virus. Hepatology 41:265–274 [CrossRef]
    [Google Scholar]
  18. Lemberg M. K., Martoglio B. 2002; Requirements for signal peptide peptidase-catalyzed intramembrane proteolysis. Mol Cell 10:735–744 [CrossRef]
    [Google Scholar]
  19. Lindenbach B. D., Evans M. J., Syder A. J., Wolk B., Tellinghuisen T. L., Liu C. C., Maruyama T., Hynes R. O., Burton D. R. other authors 2005; Complete replication of hepatitis C virus in cell culture. Science 309:623–626 [CrossRef]
    [Google Scholar]
  20. Liu Q., Tackney C., Bhat R. A., Prince A. M., Zhang P. 1997; Regulated processing of hepatitis C virus core protein is linked to subcellular localization. J Virol 71:657–662
    [Google Scholar]
  21. Lohmann V., Korner F., Koch J., Herian U., Theilmann L., Bartenschlager R. 1999; Replication of subgenomic hepatitis C virus RNAs in a hepatoma cell line. Science 285:110–113 [CrossRef]
    [Google Scholar]
  22. Major M. E., Rehermann B., Feinstone S. M. 2001; Hepatitis C viruses. In Fields Virology , 4th edn. pp 1127–1161 Edited by Knipe D. M., Howley P. M., Chanock R. M., Monath T. P., Roizman B., Straus S. E. Philadelphia, PA: Lippincott-Raven;
    [Google Scholar]
  23. McLauchlan J., Lemberg M. K., Hope G., Martoglio B. 2002; Intramembrane proteolysis promotes trafficking of hepatitis C virus core protein to lipid droplets. EMBO J 21:3980–3988 [CrossRef]
    [Google Scholar]
  24. Nakabayashi H., Taketa K., Miyano K., Yamane T., Sato J. 1982; Growth of human hepatoma cells lines with differentiated functions in chemically defined medium. Cancer Res 42:3858–3863
    [Google Scholar]
  25. Op De Beeck A., Cocquerel L., Dubuisson J. 2001; Biogenesis of hepatitis C virus envelope glycoproteins. J Gen Virol 82:2589–2595
    [Google Scholar]
  26. Owsianka A. M., Timms J. M., Tarr A. W., Brown R. J., Hickling T. P., Szwejk A., Bienkowska-Szewczyk K., Thomson B. J., Patel A. H. other authors 2006; Identification of conserved residues in the E2 envelope glycoprotein of the hepatitis C virus that are critical for CD81 binding. J Virol 80:8695–8704 [CrossRef]
    [Google Scholar]
  27. Pavlovic D., Neville D. C., Argaud O., Blumberg B., Dwek R. A., Fischer W. B., Zitzmann N. 2003; The hepatitis C virus p7 protein forms an ion channel that is inhibited by long-alkyl-chain iminosugar derivatives. Proc Natl Acad Sci U S A 100:6104–6108 [CrossRef]
    [Google Scholar]
  28. Pileri P., Uematsu Y., Campagnoli S., Galli G., Falugi F., Petracca R., Weiner A. J., Houghton M., Rosa D. other authors 1998; Binding of hepatitis C virus to CD81. Science 282:938–941 [CrossRef]
    [Google Scholar]
  29. Poynard T., Yuen M. F., Ratziu V., Lai C. L. 2003; Viral hepatitis C. Lancet 362:2095–2100 [CrossRef]
    [Google Scholar]
  30. Roccasecca R., Ansuini H., Vitelli A., Meola A., Scarselli E., Acali S., Pezzanera M., Ercole B. B., McKeating J. other authors 2003; Binding of the hepatitis C virus E2 glycoprotein to CD81 is strain specific and is modulated by a complex interplay between hypervariable regions 1 and 2. J Virol 77:1856–1867 [CrossRef]
    [Google Scholar]
  31. Rouille Y., Helle F., Delgrange D., Roingeard P., Voisset C., Blanchard E., Belouzard S., McKeating J., Patel A. H. other authors 2006; Subcellular localization of hepatitis C virus structural proteins in a cell culture system that efficiently replicates the virus. J Virol 80:2832–2841 [CrossRef]
    [Google Scholar]
  32. Roussel J., Pillez A., Montpellier C., Duverlie G., Cahour A., Dubuisson J., Wychowski C. 2003; Characterization of the expression of the hepatitis C virus F protein. J Gen Virol 84:1751–1759 [CrossRef]
    [Google Scholar]
  33. Sumpter R. Jr, Loo Y. M., Foy E., Li K., Yoneyama M., Fujita T., Lemon S. M., Gale M., Jr. 2005; Regulating intracellular antiviral defense and permissiveness to hepatitis C virus RNA replication through a cellular RNA helicase, RIG-I. J Virol 79:2689–2699 [CrossRef]
    [Google Scholar]
  34. Thomas D. L. 2000; Hepatitis C epidemiology. Curr Top Microbiol Immunol 242:25–41
    [Google Scholar]
  35. Tsukiyama-Kohara K., Iizuka N., Kohara M., Nomoto A. 1992; Internal ribosome entry site within hepatitis C virus RNA. J Virol 66:1476–1483
    [Google Scholar]
  36. Varaklioti A., Vassilaki N., Georgopoulou U., Mavromara P. 2002; Alternate translation occurs within the core coding region of the hepatitis C viral genome. J Biol Chem 277:17713–17721 [CrossRef]
    [Google Scholar]
  37. Wakita T., Pietschmann T., Kato T., Date T., Miyamoto M., Zhao Z., Murthy K., Habermann A., Krausslich H. G. other authors 2005; Production of infectious hepatitis C virus in tissue culture from a cloned viral genome. Nat Med 11:791–796 [CrossRef]
    [Google Scholar]
  38. Walewski J. L., Keller T. R., Stump D. D., Branch A. D. 2001; Evidence for a new hepatitis C virus antigen encoded in an overlapping reading frame. RNA 7:710–721 [CrossRef]
    [Google Scholar]
  39. Xu Z., Choi J., Yen T. S., Lu W., Strohecker A., Govindarajan S., Chien D., Selby M. J., Ou J. 2001; Synthesis of a novel hepatitis C virus protein by ribosomal frameshift. EMBO J 20:3840–3848 [CrossRef]
    [Google Scholar]
  40. Yanagi M., Purcell R. H., Emerson S. U., Bukh J. 1997; Transcripts from a single full-length cDNA clone of hepatitis C virus are infectious when directly transfected into the liver of a chimpanzee. Proc Natl Acad Sci U S A 94:8738–8743 [CrossRef]
    [Google Scholar]
  41. Yanagi M., Purcell R. H., Emerson S. U., Bukh J. 1999; Hepatitis C virus: an infectious molecular clone of a second major genotype (2a) and lack of viability of intertypic 1a and 2a chimeras. Virology 262:250–263 [CrossRef]
    [Google Scholar]
  42. Yasui K., Wakita T., Tsukiyama-Kohara K., Funahashi S. I., Ichikawa M., Kajita T., Moradpour D., Wands J. R., Kohara M. 1998; The native form and maturation process of hepatitis C virus core protein. J Virol 72:6048–6055
    [Google Scholar]
  43. Zhong J., Gastaminza P., Cheng G., Kapadia S., Kato T., Burton D. R., Wieland S. F., Uprichard S. L., Wakita T. other authors 2005; Robust hepatitis C virus infection in vitro. Proc Natl Acad Sci U S A 102:9294–9299 [CrossRef]
    [Google Scholar]
  44. Zhong J., Gastaminza P., Chung J., Stamataki Z., Isogawa M., Cheng G., McKeating J., Chisari F. V. 2006; Persistent hepatitis C virus infection in vitro: co-evolution of virus and host. J Virol 80:11082–11093 [CrossRef]
    [Google Scholar]
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