Skip to main content

Advertisement

SpringerLink
Log in

Negative regulation of intracellular hepatitis C virus replication by interferon regulatory factor 3

  • Published:
Journal of Gastroenterology Aims and scope Submit manuscript

Abstract

Background

Interferon regulatory factor (IRF)-3 plays an important role in initiating cellular interferon-stimulated gene-mediated antiviral responses. In the present study, we evaluated the effects of IRF-3 expression and activation on intracellular hepatitis C virus (HCV) replication using an HCV replicon system.

Methods

An HCV replicon was constructed that expressed a neomycin-selectable chimeric firefly luciferase reporter protein. A small interfering (si) RNA oligonucleotide directed against IRF-3 mRNA was designed and synthesized. A eukaryote expression plasmid vector was constructed that expressed IRF-3 mRNA under control of the cytomegalovirus early promoter/enhancer. To evaluate transcriptional activity of the interferon-stimulated genes, a reporter vector was used that expressed firefly luciferase under control of the interferon-stimulated response element (ISRE).

Results

The baseline expression of IRF-3 did not significantly differ between cells with and without expression of the replicon. Transfection of an IRF-3 expression plasmid into the cells raised the ISRE-luciferase activities. The increase of ISRE activity was significantly more potent in the replicon-expressing cells than in cells without replicon expression. Concomitantly, the overexpression of IRF-3 suppressed HCV replication levels. In contrast, siRNA knockdown of IRF-3 suppressed ISRE activity by 38% ± 2%. Interestingly, the suppression of IRF-3 resulted in a significant increase of HCV replication, by up to twofold, depending on the IRF-3 suppression levels.

Conclusions

IRF-3 negatively regulated intracellular HCV replication, and was partially activated in cells that expressed the HCV replicon. Thus, IRF-3 is a key molecule controlling HCV replication through modulation of host interferon gene responses.

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

Similar content being viewed by others

References

  1. MJ Alter (1997) ArticleTitleEpidemiology of hepatitis C Hepatology 26 62S–5S Occurrence Handle9305666 Occurrence Handle1:STN:280:ByiH2cbhvFM%3D Occurrence Handle10.1002/hep.510260711

    Article  PubMed  CAS  Google Scholar 

  2. MW Fried ML Shiffman KR Reddy C Smith G Marinos FL Goncales SuffixJr et al. (2002) ArticleTitlePeginterferon alfa-2a plus ribavirin for chronic hepatitis C virus infection N Engl J Med 347 975–82 Occurrence Handle12324553 Occurrence Handle1:CAS:528:DC%2BD38Xnt12rtb0%3D Occurrence Handle10.1056/NEJMoa020047

    Article  PubMed  CAS  Google Scholar 

  3. CE Samuel (2001) ArticleTitleAntiviral actions of interferons Clin Microbiol Rev 14 778–809 Occurrence Handle11585785 Occurrence Handle1:CAS:528:DC%2BD3MXot1Wms7o%3D Occurrence Handle10.1128/CMR.14.4.778-809.2001

    Article  PubMed  CAS  Google Scholar 

  4. T Taniguchi A Takaoka (2002) ArticleTitleThe interferon-alpha/beta system in antiviral responses: a multimodal machinery of gene regulation by the IRF family of transcription factors Curr Opin Immunol 14 111–6 Occurrence Handle11790540 Occurrence Handle1:CAS:528:DC%2BD38XktFKjsQ%3D%3D Occurrence Handle10.1016/S0952-7915(01)00305-3

    Article  PubMed  CAS  Google Scholar 

  5. GR Stark IM Kerr BR Williams RH Silverman RD Schreiber (1998) ArticleTitleHow cells respond to interferons Annu Rev Biochem 67 227–64 Occurrence Handle9759489 Occurrence Handle1:CAS:528:DyaK1cXlsFOmsLo%3D Occurrence Handle10.1146/annurev.biochem.67.1.227

    Article  PubMed  CAS  Google Scholar 

  6. CB Bigger KM Brasky RE Lanford (2001) ArticleTitleDNA microarray analysis of chimpanzee liver during acute resolving hepatitis C virus infection J Virol 75 7059–66 Occurrence Handle11435586 Occurrence Handle1:CAS:528:DC%2BD3MXlsVCgsrc%3D Occurrence Handle10.1128/JVI.75.15.7059-7066.2001

    Article  PubMed  CAS  Google Scholar 

  7. JE Darnell SuffixJr IM Kerr GR Stark (1994) ArticleTitleJak-STAT pathways and transcriptional activation in response to IFNs and other extracellular signaling proteins Science 264 1415–21 Occurrence Handle8197455 Occurrence Handle1:CAS:528:DyaK2cXktlalsL4%3D

    PubMed  CAS  Google Scholar 

  8. T Taniguchi K Ogasawara A Takaoka N Tanaka (2001) ArticleTitleIRF family of transcription factors as regulators of host defense Annu Rev Immunol 19 623–55 Occurrence Handle11244049 Occurrence Handle1:CAS:528:DC%2BD3MXivFKgu78%3D Occurrence Handle10.1146/annurev.immunol.19.1.623

    Article  PubMed  CAS  Google Scholar 

  9. M Yoneyama W Suhara Y Fukuhara M Fukuda E Nishida T Fujita (1998) ArticleTitleDirect triggering of the type I interferon system by virus infection: activation of a transcription factor complex containing IRF-3 and CBP/p300 EMBO J 17 1087–95 Occurrence Handle9463386 Occurrence Handle1:CAS:528:DyaK1cXhs1yrur0%3D Occurrence Handle10.1093/emboj/17.4.1087

    Article  PubMed  CAS  Google Scholar 

  10. R Lin C Heylbroeck PM Pitha J Hiscott (1998) ArticleTitleVirus-dependent phosphorylation of the IRF-3 transcription factor regulates nuclear translocation, transactivation potential, and proteasome-mediated degradation Mol Cell Biol 18 2986–96 Occurrence Handle9566918 Occurrence Handle1:CAS:528:DyaK1cXis1Gms70%3D

    PubMed  CAS  Google Scholar 

  11. M Sato H Suemori N Hata M Asagiri K Ogasawara K Nakao et al. (2000) ArticleTitleDistinct and essential roles of transcription factors IRF-3 and IRF-7 in response to viruses for IFN-alpha/beta gene induction Immunity 13 539–48 Occurrence Handle11070172 Occurrence Handle1:CAS:528:DC%2BD3cXnsl2nt78%3D Occurrence Handle10.1016/S1074-7613(00)00053-4

    Article  PubMed  CAS  Google Scholar 

  12. S Doyle S Vaidya R O'Connell H Dadgostar P Dempsey T Wu et al. (2002) ArticleTitleIRF3 mediates a TLR3/TLR4-specific antiviral gene program Immunity 17 251–63 Occurrence Handle12354379 Occurrence Handle1:CAS:528:DC%2BD38XnsFGrs7g%3D Occurrence Handle10.1016/S1074-7613(02)00390-4

    Article  PubMed  CAS  Google Scholar 

  13. T Nakaya M Sato N Hata M Asagiri H Suemori S Noguchi et al. (2001) ArticleTitleGene induction pathways mediated by distinct IRFs during viral infection Biochem Biophys Res Commun 283 1150–6 Occurrence Handle11355893 Occurrence Handle1:CAS:528:DC%2BD3MXjsFWhtb8%3D Occurrence Handle10.1006/bbrc.2001.4913

    Article  PubMed  CAS  Google Scholar 

  14. CF Basler A Mikulasova L Martinez-Sobrido J Paragas E Muhlberger M Bray et al. (2003) ArticleTitleThe Ebola virus VP35 protein inhibits activation of interferon regulatory factor 3 J Virol 77 7945–56 Occurrence Handle12829834 Occurrence Handle1:CAS:528:DC%2BD3sXlt1Kktbs%3D Occurrence Handle10.1128/JVI.77.14.7945-7956.2003

    Article  PubMed  CAS  Google Scholar 

  15. M Schweizer E Peterhans (2001) ArticleTitleNoncytopathic bovine viral diarrhea virus inhibits double-stranded RNA-induced apoptosis and interferon synthesis J Virol 75 4692–8 Occurrence Handle11312340 Occurrence Handle1:STN:280:DC%2BD3M3it1ehsQ%3D%3D Occurrence Handle10.1128/JVI.75.10.4692-4698.2001

    Article  PubMed  CAS  Google Scholar 

  16. J Talon CM Horvath R Polley CF Basler T Muster P Palese et al. (2000) ArticleTitleActivation of interferon regulatory factor 3 is inhibited by the influenza A virus NS1 protein J Virol 74 7989–96 Occurrence Handle10933707 Occurrence Handle1:CAS:528:DC%2BD3cXlsl2gu7c%3D Occurrence Handle10.1128/JVI.74.17.7989-7996.2000

    Article  PubMed  CAS  Google Scholar 

  17. Y He MG Katze (2002) ArticleTitleTo interfere and to anti-interfere: the interplay between hepatitis C virus and interferon Viral Immunol 15 95–119 Occurrence Handle11952150 Occurrence Handle1:CAS:528:DC%2BD38XislGgsLY%3D Occurrence Handle10.1089/088282402317340260

    Article  PubMed  CAS  Google Scholar 

  18. E Foy K Li C Wang R Sumpter SuffixJr M Ikeda SM Lemon et al. (2003) ArticleTitleRegulation of interferon regulatory factor-3 by the hepatitis C virus serine protease Science 300 1145–8 Occurrence Handle12702807 Occurrence Handle1:CAS:528:DC%2BD3sXjslGqtro%3D Occurrence Handle10.1126/science.1082604

    Article  PubMed  CAS  Google Scholar 

  19. V Lohmann F Koerner J-O Koch U Herian L Theilmann R Bartenschlager (1999) ArticleTitleReplication of subgenomic hepatitis C virus RNAs in a hepatoma cell line Science 285 110–3 Occurrence Handle10390360 Occurrence Handle1:CAS:528:DyaK1MXksVCqurk%3D Occurrence Handle10.1126/science.285.5424.110

    Article  PubMed  CAS  Google Scholar 

  20. KJ Blight AA Kolykhalov CM Rice (2000) ArticleTitleEfficient initiation of HCV RNA replication in cell culture Science 290 1972–4 Occurrence Handle11110665 Occurrence Handle1:CAS:528:DC%2BD3cXoslKgsL4%3D Occurrence Handle10.1126/science.290.5498.1972

    Article  PubMed  CAS  Google Scholar 

  21. M Frese V Schwarzle K Barth N Krieger V Lohmann S Mihm et al. (2002) ArticleTitleInterferon-gamma inhibits replication of subgenomic and genomic hepatitis C virus RNAs Hepatology 35 694–703 Occurrence Handle11870386 Occurrence Handle1:CAS:528:DC%2BD38Xitlyqtr8%3D Occurrence Handle10.1053/jhep.2002.31770

    Article  PubMed  CAS  Google Scholar 

  22. JT Guo VV Bichko C Seeger (2001) ArticleTitleEffect of alpha interferon on the hepatitis C virus replicon J Virol 75 8516–23 Occurrence Handle11507197 Occurrence Handle1:CAS:528:DC%2BD3MXmsVaru7g%3D Occurrence Handle10.1128/JVI.75.18.8516-8523.2001

    Article  PubMed  CAS  Google Scholar 

  23. N Kanazawa M Kurosaki N Sakamoto N Enomoto Y Itsui T Yamashiro et al. (2004) ArticleTitleRegulation of hepatitis C virus replication by interferon regulatory factor-1 J Virol 78 9713–20 Occurrence Handle15331704 Occurrence Handle1:CAS:528:DC%2BD2cXnvVaku7Y%3D Occurrence Handle10.1128/JVI.78.18.9713-9720.2004

    Article  PubMed  CAS  Google Scholar 

  24. T Yokota N Sakamoto N Enomoto Y Tanabe M Miyagishi S Maekawa et al. (2003) ArticleTitleInhibition of intracellular hepatitis C virus replication by synthetic and vector-derived small interfering RNAs EMBO Rep 4 602–8 Occurrence Handle12740604 Occurrence Handle1:CAS:528:DC%2BD3sXktFOgtr0%3D Occurrence Handle10.1038/sj.embor.embor840

    Article  PubMed  CAS  Google Scholar 

  25. Y Tanabe N Sakamoto N Enomoto M Kurosaki E Ueda S Maekawa et al. (2004) ArticleTitleSynergistic inhibition of intracellular hepatitis C virus replication by combination of ribavirin and interferon-alpha J Infect Dis 189 1129–39 Occurrence Handle15031779 Occurrence Handle1:CAS:528:DC%2BD2cXjsleqsrs%3D Occurrence Handle10.1086/382595

    Article  PubMed  CAS  Google Scholar 

  26. S Oshima T Nakamura S Namiki E Okada K Tsuchiya R Okamoto et al. (2004) ArticleTitleInterferon regulatory factor 1 (IRF-1) and IRF-2 distinctively up-regulate gene expression and production of interleukin-7 in human intestinal epithelial cells Mol Cell Biol 24 6298–310 Occurrence Handle15226432 Occurrence Handle1:CAS:528:DC%2BD2cXlslensbw%3D Occurrence Handle10.1128/MCB.24.14.6298-6310.2004

    Article  PubMed  CAS  Google Scholar 

  27. K Takeda T Kaisho S Akira (2003) ArticleTitleToll-like receptors Annu Rev Immunol 21 335–76 Occurrence Handle12524386 Occurrence Handle1:CAS:528:DC%2BD3sXjtF2isrg%3D Occurrence Handle10.1146/annurev.immunol.21.120601.141126

    Article  PubMed  CAS  Google Scholar 

  28. KA Fitzgerald SM McWhirter KL Faia DC Rowe E Latz DT Golenbock et al. (2003) ArticleTitleIKKepsilon and TBK1 are essential components of the IRF3 signaling pathway Nat Immunol 4 491–6 Occurrence Handle12692549 Occurrence Handle1:CAS:528:DC%2BD3sXjtlakt7k%3D Occurrence Handle10.1038/ni921

    Article  PubMed  CAS  Google Scholar 

  29. S Sharma BR tenOever N Grandvaux GP Zhou R Lin J Hiscott (2003) ArticleTitleTriggering the interferon antiviral response through an IKK-related pathway Science 300 1148–51 Occurrence Handle12702806 Occurrence Handle1:CAS:528:DC%2BD3sXjslGqtrs%3D Occurrence Handle10.1126/science.1081315

    Article  PubMed  CAS  Google Scholar 

  30. M Yamamoto S Sato K Mori K Hoshino O Takeuchi K Takeda et al. (2002) ArticleTitleCutting edge: a novel Toll/IL-1 receptor domain-containing adapter that preferentially activates the IFN-beta promoter in the Toll-like receptor signaling J Immunol 169 6668–72 Occurrence Handle12471095 Occurrence Handle1:CAS:528:DC%2BD38XpsFCqu74%3D

    PubMed  CAS  Google Scholar 

  31. RE Lanford B Guerra H Lee DR Averett B Pfeiffer D Chavez et al. (2003) ArticleTitleAntiviral effect and virus-host interactions in response to alpha interferon, gamma interferon, poly(i)-poly(c), tumor necrosis factor alpha, and ribavirin in hepatitis C virus subgenomic replicons J Virol 77 1092–104 Occurrence Handle12502825 Occurrence Handle1:CAS:528:DC%2BD3sXitl2htQ%3D%3D Occurrence Handle10.1128/JVI.77.2.1092-1104.2003

    Article  PubMed  CAS  Google Scholar 

  32. M Yoneyama M Kikuchi T Natsukawa N Shinobu T Imaizumi M Miyagishi et al. (2004) ArticleTitleThe RNA helicase RIG-I has an essential function in double-stranded RNA-induced innate antiviral responses Nat Immunol 5 730–7 Occurrence Handle15208624 Occurrence Handle1:CAS:528:DC%2BD2cXlt1Wmu78%3D Occurrence Handle10.1038/ni1087

    Article  PubMed  CAS  Google Scholar 

  33. Y Castelruiz E Larrea P Boya MP Civeira J Prieto (1999) ArticleTitleInterferon alpha subtypes and levels of type I interferons in the liver and peripheral mononuclear cells in patients with chronic hepatitis C and controls Hepatology 29 1900–4 Occurrence Handle10347136 Occurrence Handle1:CAS:528:DyaK1MXjvFOmsbo%3D Occurrence Handle10.1002/hep.510290625

    Article  PubMed  CAS  Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Department of Gastroenterology and Hepatology, Tokyo Medical and Dental University, 1-5-45 Yushima, Bunkyo-ku, Tokyo, 113-8519, Japan

    Tsuyoshi Yamashiro, Naoya Sakamoto, Masayuki Kurosaki, Nobuhiko Kanazawa, Yoko Tanabe, Mina Nakagawa, Cheng-Hsin Chen, Yasuhiro Itsui, Tomoyuki Koyama, Yoshie Takeda, Shinya Maekawa & Mamoru Watanabe

  2. First Department of Internal Medicine, University of Yamanashi, Chuo, Japan

    Shinya Maekawa & Nobuyuki Enomoto

  3. First Department of Internal Medicine, School of Medicine, University of the Ryukyus, Okinawa, Japan

    Tsuyoshi Yamashiro & Hiroshi Sakugawa

Authors
  1. Tsuyoshi Yamashiro
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Naoya Sakamoto
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Masayuki Kurosaki
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Nobuhiko Kanazawa
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Yoko Tanabe
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Mina Nakagawa
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Cheng-Hsin Chen
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. Yasuhiro Itsui
    View author publications

    You can also search for this author in PubMed Google Scholar

  9. Tomoyuki Koyama
    View author publications

    You can also search for this author in PubMed Google Scholar

  10. Yoshie Takeda
    View author publications

    You can also search for this author in PubMed Google Scholar

  11. Shinya Maekawa
    View author publications

    You can also search for this author in PubMed Google Scholar

  12. Nobuyuki Enomoto
    View author publications

    You can also search for this author in PubMed Google Scholar

  13. Hiroshi Sakugawa
    View author publications

    You can also search for this author in PubMed Google Scholar

  14. Mamoru Watanabe
    View author publications

    You can also search for this author in PubMed Google Scholar

Rights and permissions

Reprints and permissions

About this article

Cite this article

Yamashiro, T., Sakamoto, N., Kurosaki, M. et al. Negative regulation of intracellular hepatitis C virus replication by interferon regulatory factor 3. J Gastroenterol 41, 750–757 (2006). https://doi.org/10.1007/s00535-006-1842-x

Download citation

  • Received:

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s00535-006-1842-x

Key words

Search

Navigation