1887

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Volume 96, Issue 2

Regulation of core expression during the hepatitis C virus life cycle Free

Abstract

Core plays a critical role during hepatitis C virus (HCV) assembly, not only as a structural component of the virion, but also as a regulator of the formation of assembly sites. In this study, we observed that core is expressed later than other HCV proteins in a single viral cycle assay, resulting in a relative increase of core expression during a late step of the viral life cycle. This delayed core expression results from an increase of core half-life, indicating that core is initially degraded and is stabilized at a late step of the HCV life cycle. Stabilization-mediated delayed kinetics of core expression were also observed using heterologous expression systems. Core stabilization did not depend on its interaction with non-structural proteins or lipid droplets but was correlated with its expression levels and its oligomerization status. Therefore in the course of a HCV infection, core stabilization is likely to occur when the prior amplification of the viral genome during an initial replication step allows core to be synthesized at higher levels as a stable protein, during the assembly step of the viral life cycle.

  • Received:
  • Accepted:
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© 2015 The Authors
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2015-02-01
2024-05-03
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References

  1. Alsaleh K., Delavalle P.-Y., Pillez A., Duverlie G., Descamps V., Rouillé Y., Dubuisson J., Wychowski C. 2010; Identification of basic amino acids at the N-terminal end of the core protein that are crucial for hepatitis C virus infectivity. J Virol 84:12515–12528 [View Article][PubMed]
     [Google Scholar]
  2. Bartenschlager R., Penin F., Lohmann V., André P. 2011; Assembly of infectious hepatitis C virus particles. Trends Microbiol 19:95–103 [View Article][PubMed]
     [Google Scholar]
  3. Boulant S., Vanbelle C., Ebel C., Penin F., Lavergne J.-P. 2005; Hepatitis C virus core protein is a dimeric alpha-helical protein exhibiting membrane protein features. J Virol 79:11353–11365 [View Article][PubMed]
     [Google Scholar]
  4. Boulant S., Targett-Adams P., McLauchlan J. 2007; Disrupting the association of hepatitis C virus core protein with lipid droplets correlates with a loss in production of infectious virus. J Gen Virol 88:2204–2213 [View Article][PubMed]
     [Google Scholar]
  5. Clayton R. F., Owsianka A., Aitken J., Graham S., Bhella D., Patel A. H. 2002; Analysis of antigenicity and topology of E2 glycoprotein present on recombinant hepatitis C virus-like particles. J Virol 76:7672–7682 [View Article][PubMed]
     [Google Scholar]
  6. Dentzer T. G., Lorenz I. C., Evans M. J., Rice C. M. 2009; Determinants of the hepatitis C virus nonstructural protein 2 protease domain required for production of infectious virus. J Virol 83:12702–12713 [View Article][PubMed]
     [Google Scholar]
  7. Descamps V., Op de Beeck A., Plassart C., Brochot E., François C., Helle F., Adler M., Bourgeois N., Degré D. et al. 2012; Strong correlation between liver and serum levels of hepatitis C virus core antigen and RNA in chronically infected patients. J Clin Microbiol 50:465–468 [View Article][PubMed]
     [Google Scholar]
  8. Dubuisson J., Hsu H. H., Cheung R. C., Greenberg H. B., Russell D. G., Rice C. M. 1994; Formation and intracellular localization of hepatitis C virus envelope glycoprotein complexes expressed by recombinant vaccinia and Sindbis viruses. J Virol 68:6147–6160[PubMed]
     [Google Scholar]
  9. Eng F. J., Walewski J. L., Klepper A. L., Fishman S. L., Desai S. M., McMullan L. K., Evans M. J., Rice C. M., Branch A. D. 2009; Internal initiation stimulates production of p8 minicore, a member of a newly discovered family of hepatitis C virus core protein isoforms. J Virol 83:3104–3114 [View Article][PubMed]
     [Google Scholar]
  10. 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[PubMed]
     [Google Scholar]
  11. Goueslain L., Alsaleh K., Horellou P., Roingeard P., Descamps V., Duverlie G., Ciczora Y., Wychowski C., Dubuisson J., Rouillé Y. 2010; Identification of GBF1 as a cellular factor required for hepatitis C virus RNA replication. J Virol 84:773–787 [View Article][PubMed]
     [Google Scholar]
  12. Hope R. G., McLauchlan J. 2000; Sequence motifs required for lipid droplet association and protein stability are unique to the hepatitis C virus core protein. J Gen Virol 81:1913–1925[PubMed]
     [Google Scholar]
  13. Jirasko V., Montserret R., Lee J. Y., Gouttenoire J., Moradpour D., Penin F., Bartenschlager R. 2010; Structural and functional studies of nonstructural protein 2 of the hepatitis C virus reveal its key role as organizer of virion assembly. PLoS Pathog 6:e1001233 [View Article][PubMed]
     [Google Scholar]
  14. Kato T., Date T., Miyamoto M., Furusaka A., Tokushige K., Mizokami M., Wakita T. 2003; Efficient replication of the genotype 2a hepatitis C virus subgenomic replicon. Gastroenterology 125:1808–1817 [View Article][PubMed]
     [Google Scholar]
  15. Klein K. C., Polyak S. J., Lingappa J. R. 2004; Unique features of hepatitis C virus capsid formation revealed by de novo cell-free assembly. J Virol 78:9257–9269 [View Article][PubMed]
     [Google Scholar]
  16. Kunkel M., Lorinczi M., Rijnbrand R., Lemon S. M., Watowich S. J. 2001; Self-assembly of nucleocapsid-like particles from recombinant hepatitis C virus core protein. J Virol 75:2119–2129 [View Article][PubMed]
     [Google Scholar]
  17. Lavanchy D. 2011; Evolving epidemiology of hepatitis C virus. Clin Microbiol Infect 17:107–115 [View Article][PubMed]
     [Google Scholar]
  18. Lindenbach B. D., Evans M. J., Syder A. J., Wölk B., Tellinghuisen T. L., Liu C. C., Maruyama T., Hynes R. O., Burton D. R. et al. 2005; Complete replication of hepatitis C virus in cell culture. Science 309:623–626 [View Article][PubMed]
     [Google Scholar]
  19. Lindenbach B. D., Thiel H.-J., Rice C. M. 2007; Flaviviridae: the viruses and their replication. In Fields Virology, 5th edn. pp. 1101–1152 Edited by Knipe D. M., Howley P. M. Philadelphia, PA: Lippincott, Williams & Wilkins;
     [Google Scholar]
  20. Lohmann V., Körner 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 [View Article][PubMed]
     [Google Scholar]
  21. Macdonald A., Crowder K., Street A., McCormick C., Saksela K., Harris M. 2003; The hepatitis C virus non-structural NS5A protein inhibits activating protein-1 function by perturbing ras-ERK pathway signaling. J Biol Chem 278:17775–17784 [View Article][PubMed]
     [Google Scholar]
  22. Maillard P., Krawczynski K., Nitkiewicz J., Bronnert C., Sidorkiewicz M., Gounon P., Dubuisson J., Faure G., Crainic R., Budkowska A. 2001; Nonenveloped nucleocapsids of hepatitis C virus in the serum of infected patients. J Virol 75:8240–8250 [View Article][PubMed]
     [Google Scholar]
  23. Majeau N., Gagné V., Boivin A., Bolduc M., Majeau J.-A., Ouellet D., Leclerc D. 2004; The N-terminal half of the core protein of hepatitis C virus is sufficient for nucleocapsid formation. J Gen Virol 85:971–981 [View Article][PubMed]
     [Google Scholar]
  24. Masaki T., Suzuki R., Murakami K., Aizaki H., Ishii K., Murayama A., Date T., Matsuura Y., Miyamura T. et al. 2008; Interaction of hepatitis C virus nonstructural protein 5A with core protein is critical for the production of infectious virus particles. J Virol 82:7964–7976 [View Article][PubMed]
     [Google Scholar]
  25. Maurin G., Fresquet J., Granio O., Wychowski C., Cosset F.-L., Lavillette D. 2011; Identification of interactions in the E1E2 heterodimer of hepatitis C virus important for cell entry. J Biol Chem 286:23865–23876 [View Article][PubMed]
     [Google Scholar]
  26. 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 [View Article][PubMed]
     [Google Scholar]
  27. McMullan L. K., Grakoui A., Evans M. J., Mihalik K., Puig M., Branch A. D., Feinstone S. M., Rice C. M. 2007; Evidence for a functional RNA element in the hepatitis C virus core gene. Proc Natl Acad Sci U S A 104:2879–2884 [View Article][PubMed]
     [Google Scholar]
  28. Miyanari Y., Atsuzawa K., Usuda N., Watashi K., Hishiki T., Zayas M., Bartenschlager R., Wakita T., Hijikata M., Shimotohno K. 2007; The lipid droplet is an important organelle for hepatitis C virus production. Nat Cell Biol 9:1089–1097 [View Article][PubMed]
     [Google Scholar]
  29. Murray C. L., Jones C. T., Tassello J., Rice C. M. 2007; Alanine scanning of the hepatitis C virus core protein reveals numerous residues essential for production of infectious virus. J Virol 81:10220–10231 [View Article][PubMed]
     [Google Scholar]
  30. 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[PubMed]
     [Google Scholar]
  31. Popescu C.-I., Callens N., Trinel D., Roingeard P., Moradpour D., Descamps V., Duverlie G., Penin F., Héliot L. et al. 2011; NS2 protein of hepatitis C virus interacts with structural and non-structural proteins towards virus assembly. PLoS Pathog 7:e1001278 [View Article][PubMed]
     [Google Scholar]
  32. Rocha-Perugini V., Lavie M., Delgrange D., Canton J., Pillez A., Potel J., Lecoeur C., Rubinstein E., Dubuisson J. et al. 2009; The association of CD81 with tetraspanin-enriched microdomains is not essential for Hepatitis C virus entry. BMC Microbiol 9:111 [View Article][PubMed]
     [Google Scholar]
  33. Rouillé Y., Helle F., Delgrange D., Roingeard P., Voisset C., Blanchard E., Belouzard S., McKeating J., Patel A. H. et al. 2006; Subcellular localization of hepatitis C virus structural proteins in a cell culture system that efficiently replicates the virus. J Virol 80:2832–2841 [View Article][PubMed]
     [Google Scholar]
  34. Séron K., Couturier C., Belouzard S., Bacart J., Monté D., Corset L., Bocquet O., Dam J., Vauthier V. et al. 2011; Endospanins regulate a postinternalization step of the leptin receptor endocytic pathway. J Biol Chem 286:17968–17981 [View Article][PubMed]
     [Google Scholar]
  35. Shi S. T., Lee K.-J., Aizaki H., Hwang S. B., Lai M. M. C. 2003; Hepatitis C virus RNA replication occurs on a detergent-resistant membrane that cofractionates with caveolin-2. J Virol 77:4160–4168 [View Article][PubMed]
     [Google Scholar]
  36. Shimoike T., Koyama C., Murakami K., Suzuki R., Matsuura Y., Miyamura T., Suzuki T. 2006; Down-regulation of the internal ribosome entry site (IRES)-mediated translation of the hepatitis C virus: critical role of binding of the stem-loop IIId domain of IRES and the viral core protein. Virology 345:434–445 [View Article][PubMed]
     [Google Scholar]
  37. Shirakura M., Murakami K., Ichimura T., Suzuki R., Shimoji T., Fukuda K., Abe K., Sato S., Fukasawa M. et al. 2007; E6AP ubiquitin ligase mediates ubiquitylation and degradation of hepatitis C virus core protein. J Virol 81:1174–1185 [View Article][PubMed]
     [Google Scholar]
  38. Stapleford K. A., Lindenbach B. D. 2011; Hepatitis C virus NS2 coordinates virus particle assembly through physical interactions with the E1-E2 glycoprotein and NS3-NS4A enzyme complexes. J Virol 85:1706–1717 [View Article][PubMed]
     [Google Scholar]
  39. Suzuki R., Tamura K., Li J., Ishii K., Matsuura Y., Miyamura T., Suzuki T. 2001; Ubiquitin-mediated degradation of hepatitis C virus core protein is regulated by processing at its carboxyl terminus. Virology 280:301–309 [View Article][PubMed]
     [Google Scholar]
  40. Vassilaki N., Friebe P., Meuleman P., Kallis S., Kaul A., Paranhos-Baccalà G., Leroux-Roels G., Mavromara P., Bartenschlager R. 2008; Role of the hepatitis C virus core+1 open reading frame and core cis-acting RNA elements in viral RNA translation and replication. J Virol 82:11503–11515 [View Article][PubMed]
     [Google Scholar]
  41. Wakita T., Pietschmann T., Kato T., Date T., Miyamoto M., Zhao Z., Murthy K., Habermann A., Kräusslich H.-G. et al. 2005; Production of infectious hepatitis C virus in tissue culture from a cloned viral genome. Nat Med 11:791–796 [View Article][PubMed]
     [Google Scholar]
  42. 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 [View Article][PubMed]
     [Google Scholar]
  43. 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 [View Article][PubMed]
     [Google Scholar]
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Regulation of core expression during the hepatitis C virus life cycle
J. Gen. Virol. 96, 311 (2015); https://doi.org/10.1099/vir.0.070433-0
/content/journal/jgv/10.1099/vir.0.070433-0
/content/journal/jgv/10.1099/vir.0.070433-0
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