Skip to main content

Advertisement

SpringerLink
Log in

Hepatitis C virus NS2/3 protease regulates HCV IRES-dependent translation and NS5B RdRp activity

  • Original Article
  • Published:
Archives of Virology Aims and scope Submit manuscript

Abstract

Chronic hepatitis C virus (HCV) infection often leads to liver cancer. NS2/3 protease is the first of two virally encoded proteases required for HCV polyprotein processing. In this report, we investigated the function of NS2/3 protease on HCV replication and translation. Cells transfected with plasmids encoding wild-type or mutant NS2/3 and a dual-luciferase reporter construct containing an HCV internal ribosome entry site (IRES) were used to examine the effect of NS2/3 protease on translation of HCV RNA. Cells transfected with plasmids encoding wild-type or mutant NS2/3, pcDNA-NS5B and a reporter plasmid were used to examine the effect of NS2/3 protease on HCV replication. The results showed that both autocleavage processing and the uncleaved form of NS2/3 protease specifically decrease HCV IRES-directed translation, while the uncleaved form of NS2/3 protease decreases HCV NS5B RdRp activity (replication), indicating that autoregulation by NS2/3 protease of HCV replication and translation may play an important role in persistent HCV infection.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4

Similar content being viewed by others

References

  1. Ali N, Siddiqui A (1995) Interaction of polypyrimidine tract-binding protein with the 5′ noncoding region of the hepatitis C virus RNA genome and its functional requirement in internal initiation of translation. J Virol 69:6367–6375

    PubMed  CAS  Google Scholar 

  2. Alter MJ (1997) Epidemiology of hepatitis C. Hepatology 26:62S–65S

    Article  PubMed  CAS  Google Scholar 

  3. Behrens SE, Tomei L, De Francesco R (1996) Identification and properties of the RNA-dependent RNA polymerase of hepatitis C virus. Embo J 15:12–22

    PubMed  CAS  Google Scholar 

  4. Bukh J, Purcell RH, Miller RH (1992) Sequence analysis of the 5′ noncoding region of hepatitis C virus. Proc Natl Acad Sci USA 89:4942–4946

    Article  PubMed  CAS  Google Scholar 

  5. Date T, Kato T, Miyamoto M, Zhao Z, Yasui K, Mizokami M, Wakita T (2004) Genotype 2a hepatitis C virus subgenomic replicon can replicate in HepG2 and IMY-N9 cells. J Biol Chem 279:22371–22376

    Article  PubMed  CAS  Google Scholar 

  6. Friebe P, Lohmann V, Krieger N, Bartenschlager R (2001) Sequences in the 5′ nontranslated region of hepatitis C virus required for RNA replication. J Virol 75:12047–12057

    Article  PubMed  CAS  Google Scholar 

  7. Gale M Jr, Tan SL, Katze MG (2000) Translational control of viral gene expression in eukaryotes. Microbiol Mol Biol Rev 64:239–280

    Article  PubMed  CAS  Google Scholar 

  8. He Y, Yan W, Coito C, Li Y Jr, Gale M, Katze MG (2003) The regulation of hepatitis C virus (HCV) internal ribosome-entry site-mediated translation by HCV replicons and nonstructural proteins. J Gen Virol 84:535–543

    Article  PubMed  CAS  Google Scholar 

  9. Hellen CU, Pestova TV (1999) Tranlation of hepatitis C virus RNA. J Viral Hepat 6:79–87

    Article  PubMed  CAS  Google Scholar 

  10. Ito T, Tahara SM, Lai MM (1998) The 3′-untranslated region of hepatitis C virus RNA enhances translation from an internal ribosomal entry site. J Virol 72:8789–8796

    PubMed  CAS  Google Scholar 

  11. Izumi RE, Das S, Barat B, Raychaudhuri S, Dasgupta A (2004) A peptide from autoantigen La blocks poliovirus and hepatitis C virus cap-independent translation and reveals a single tyrosine critical for La RNA binding and translation stimulation. J Virol 78:3763–3776

    Article  PubMed  CAS  Google Scholar 

  12. Jones CT, Murray CL, Eastman DK, Tassello J, Rice CM (2007) Hepatitis C virus p7 and NS2 proteins are essential for production of infectious virus. J Virol 81:8374–8383

    Article  PubMed  CAS  Google Scholar 

  13. Kalliampakou KI, Kalamvoki M, Mavromara P (2005) Hepatitis C virus (HCV) NS5A protein downregulates HCV IRES-dependent translation. J Gen Virol 86:1015–1025

    Article  PubMed  CAS  Google Scholar 

  14. Kato J, Kato N, Yoshida H, Ono-Nita SK, Shiratori Y, Omata M (2002) Hepatitis C virus NS4A and NS4B proteins suppress translation in vivo. J Med Virol 66:187–199

    Article  PubMed  CAS  Google Scholar 

  15. Kim YK, Lee SH, Kim CS, Seol SK, Jang SK (2003) Long-range RNA-RNA interaction between the 5′ nontranslated region and the core-coding sequences of hepatitis C virus modulates the IRES-dependent translation. RNA 9:599–606

    Article  PubMed  CAS  Google Scholar 

  16. Kolykhalov AA, Mihalik K, Feinstone SM, Rice CM (2000) Hepatitis C virus-encoded enzymatic activities and conserved RNA elements in the 3′ nontranslated region are essential for virus replication in vivo. J Virol 74:2046–2051

    Article  PubMed  CAS  Google Scholar 

  17. Li D, Takyar ST, Lott WB, Gowans EJ (2003) Amino acids 1–20 of the hepatitis C virus (HCV) core protein specifically inhibit HCV IRES-dependent translation in HepG2 cells, and inhibit both HCV IRES- and cap-dependent translation in HuH7 and CV-1 cells. J Gen Virol 84:815–825

    Article  PubMed  CAS  Google Scholar 

  18. Lorenz IC, Marcotrigiano J, Dentzer TG, Rice CM (2006) Structure of the catalytic domain of the hepatitis C virus NS2–3 protease. Nature 442:831–835

    Article  PubMed  CAS  Google Scholar 

  19. Moriishi K, Matsuura Y (2007) Host factors involved in the replication of hepatitis C virus. Rev Med Virol 11:343–354

    Article  Google Scholar 

  20. Murakami K, Abe M, Kageyama T, Kamoshita N, Nomoto A (2001) Down-regulation of translation driven by hepatitis C virus internal ribosomal entry site by the 3′ untranslated region of RNA. Arch Virol 146:729–741

    Article  PubMed  CAS  Google Scholar 

  21. Pietschmann T, Lohmann V, Rutter G, Kurpanek K, Bartenschlager R (2001) Characterization of cell lines carrying self-replicating hepatitis C virus RNAs. J Virol 75:1252–1264

    Article  PubMed  CAS  Google Scholar 

  22. Reed KE, Rice CM (2000) Overview of hepatitis C virus genome structure, polyprotein processing, and protein properties. Curr Top Microbiol Immunol 242:55–84

    PubMed  CAS  Google Scholar 

  23. She Y, Liao Q, Chen X, Ye L, Wu Z (2008) Hepatitis C virus (HCV) NS2 protein up-regulates HCV IRES-dependent translation and down-regulates NS5B RdRp activity. Arch Virol 153:1991–1997

    Article  PubMed  CAS  Google Scholar 

  24. Shimoike T, Mimori S, Tani H, Matsuura Y, Miyamura T (1999) Interaction of hepatitis C virus core protein with viral sense RNA and suppression of its translation. J Virol 73:9718–9725

    PubMed  CAS  Google Scholar 

  25. Thompson SR, Sarnow P (2000) Regulation of host cell translation by viruses and effects on cell function. Curr Opin Microbiol 3:366–370

    Article  PubMed  CAS  Google Scholar 

  26. Wang TH, Rijnbrand RC, Lemon SM (2000) Core protein-coding sequence, but not core protein, modulates the efficiency of cap-independent translation directed by the internal ribosome entry site of hepatitis C virus. J Virol 74:11347–11358

    Article  PubMed  CAS  Google Scholar 

  27. Welbourn S, Green R, Gamache I, Dandache S, Lohmann V, Bartenschlager R, Meerovitch K, Pause A (2005) Hepatitis C virus NS2/3 processing is required for NS3 stability and viral RNA replication. J Biol Chem 280:29604–29611

    Article  PubMed  CAS  Google Scholar 

  28. Welbourn S, Pause A (2007) The hepatitis C virus NS2/3 protease. Curr Issues Mol Biol 9:63–69

    PubMed  CAS  Google Scholar 

Download references

Acknowledgments

We thank Charles M. Rice from Washington University for providing plasmid Flag-tagged HCV NS2-3 H143A. We are grateful to Dr. Chen Xulin from Wuhan Institute of Virology (CAS) and He Biao from the Pennsylvania State University for proofreading the manuscript and helpful discussions.

Author information

Authors and Affiliations

  1. State Key Laboratory of Virology, College of Life Science, Wuhan Univerisity, 430072, Wuhan, Hubei, Poeple’s Republic of China

    Yinglong She, Tao Han, Linbai Ye & Zhenghui Wu

Authors
  1. Yinglong She
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Tao Han
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Linbai Ye
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Zhenghui Wu
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Linbai Ye.

Rights and permissions

Reprints and permissions

About this article

Cite this article

She, Y., Han, T., Ye, L. et al. Hepatitis C virus NS2/3 protease regulates HCV IRES-dependent translation and NS5B RdRp activity. Arch Virol 154, 1465–1473 (2009). https://doi.org/10.1007/s00705-009-0469-7

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s00705-009-0469-7

Keywords

Search

Navigation