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Abstract

Hepatitis B virus (HBV) infection remains one of the most serious health problems worldwide. Whilst studies have shown that HBV impairs interferon (IFN) production from dendritic cells in chronic hepatitis B patients, it remains unknown whether HBV inhibits IFN production in human hepatocytes. Using transient transfection assays in a primary human hepatocyte cell line (PH5CH8), this study demonstrated that HBV polymerase inhibits promoter activity induced by Newcastle disease virus, Sendai virus or poly(I : C) in a dose-dependent manner, whilst ectopic expression of the HBV core and X proteins had no effect on promoter activity. In addition, HBV polymerase blocked cellular IFN- expression and consequent antiviral immunity revealed by an infection protection assay. Furthermore, overexpression of key molecules on the IFN- induction axis, together with HBV polymerase, resulted in a block of promoter activity triggered by RIG-I, IPS-1, TRIF, TBK1 and IKKϵ, but not by an IFN regulatory factor 3 dominant-positive mutant (IRF3-5D), suggesting that HBV polymerase prevents IFN- expression at the TBK1/IKKϵ level. Further studies showed that HBV polymerase inhibited phosphorylation, dimerization and nuclear translocation of IRF3, in response to Sendai virus infection. Finally, it was shown that HBV polymerase-mediated dampening of the interaction between TBK1/IKKϵ and DDX3 may be involved in the inhibitory effect on IFN- induction. Taken together, these findings reveal a novel role of HBV polymerase in HBV counteraction of IFN- production in human hepatocytes.

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2010-08-01
2019-11-19
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References

  1. Ablasser, A., Bauernfeind, F., Hartmann, G., Latz, E., Fitzgerald, K. A. & Hornung, V. ( 2009; ). RIG-I-dependent sensing of poly(dA : dT) through the induction of an RNA polymerase III-transcribed RNA intermediate. Nat Immunol 10, 1065–1072.[CrossRef]
    [Google Scholar]
  2. Basler, C. F., Mikulasova, A., Martinez-Sobrido, L., Paragas, J., Muhlberger, E., Bray, M., Klenk, H. D., Palese, P. & García-Sastre, A. ( 2003; ). The Ebola virus VP35 protein inhibits activation of interferon regulatory factor 3. J Virol 77, 7945–7956.[CrossRef]
    [Google Scholar]
  3. Cardenas, W. B., Loo, Y. M., Gale, M., Jr, Hartman, A. L., Kimberlin, C. R., Martinez-Sobrido, L., Saphire, E. O. & Basler, C. F. ( 2006; ). Ebola virus VP35 protein binds double-stranded RNA and inhibits alpha/beta interferon production induced by RIG-I signaling. J Virol 80, 5168–5178.[CrossRef]
    [Google Scholar]
  4. Chiu, Y. H., Macmillan, J. B. & Chen, Z. J. ( 2009; ). RNA polymerase III detects cytosolic DNA and induces type I interferons through the RIG-I pathway. Cell 138, 576–591.[CrossRef]
    [Google Scholar]
  5. Crispe, I. N. ( 2009; ). The liver as a lymphoid organ. Annu Rev Immunol 27, 147–163.[CrossRef]
    [Google Scholar]
  6. Ferreon, J. C., Ferreon, A. C., Li, K. & Lemon, S. M. ( 2005; ). Molecular determinants of TRIF proteolysis mediated by the hepatitis C virus NS3/4A protease. J Biol Chem 280, 20483–20492.[CrossRef]
    [Google Scholar]
  7. Foster, G. R., Ackrill, A. M., Goldin, R. D., Kerr, I. M., Thomas, H. C. & Stark, G. R. ( 1991; ). Expression of the terminal protein region of hepatitis B virus inhibits cellular responses to interferons alpha and gamma and double-stranded RNA. Proc Natl Acad Sci U S A 88, 2888–2892.[CrossRef]
    [Google Scholar]
  8. Ganem, D. & Prince, A. M. ( 2004; ). Hepatitis B virus infection – natural history and clinical consequences. N Engl J Med 350, 1118–1129.[CrossRef]
    [Google Scholar]
  9. Hiscott, J. ( 2007; ). Triggering the innate antiviral response through IRF-3 activation. J Biol Chem 282, 15325–15329.[CrossRef]
    [Google Scholar]
  10. Iwamura, T., Yoneyama, M., Yamaguchi, K., Suhara, W., Mori, W., Shiota, K., Okabe, Y., Namiki, H. & Fujita, T. ( 2001; ). Induction of IRF-3/-7 kinase and NF-κB in response to double-stranded RNA and virus infection: common and unique pathways. Genes Cells 6, 375–388.[CrossRef]
    [Google Scholar]
  11. Kalverda, A. P., Thompson, G. S., Vogel, A., Schroder, M., Bowie, A. G., Khan, A. R. & Homans, S. W. ( 2009; ). Poxvirus K7 protein adopts a Bcl-2 fold: biochemical mapping of its interactions with human DEAD box RNA helicase DDX3. J Mol Biol 385, 843–853.[CrossRef]
    [Google Scholar]
  12. Li, K., Chen, Z., Kato, N., Gale, M., Jr & Lemon, S. M. ( 2005a; ). Distinct poly(I-C) and virus-activated signaling pathways leading to interferon-β production in hepatocytes. J Biol Chem 280, 16739–16747.[CrossRef]
    [Google Scholar]
  13. Li, K., Foy, E., Ferreon, J. C., Nakamura, M., Ferreon, A. C., Ikeda, M., Ray, S. C., Gale, M., Jr & Lemon, S. M. ( 2005b; ). Immune evasion by hepatitis C virus NS3/4A protease-mediated cleavage of the Toll-like receptor 3 adaptor protein TRIF. Proc Natl Acad Sci U S A 102, 2992–2997.[CrossRef]
    [Google Scholar]
  14. Li, X. D., Sun, L., Seth, R. B., Pineda, G. & Chen, Z. J. ( 2005c; ). Hepatitis C virus protease NS3/4A cleaves mitochondrial antiviral signaling protein off the mitochondria to evade innate immunity. Proc Natl Acad Sci U S A 102, 17717–17722.[CrossRef]
    [Google Scholar]
  15. Liaw, Y. F. & Chu, C. M. ( 2009; ). Hepatitis B virus infection. Lancet 373, 582–592.[CrossRef]
    [Google Scholar]
  16. Lucifora, J., Durantel, D., Testoni, B., Hantz, O., Levrero, M. & Zoulim, F. ( 2010; ). Control of hepatitis B virus replication by innate response of HepaRG cells. Hepatology 51, 63–72.[CrossRef]
    [Google Scholar]
  17. Meylan, E., Curran, J., Hofmann, K., Moradpour, D., Binder, M., Bartenschlager, R. & Tschopp, J. ( 2005; ). Cardif is an adaptor protein in the RIG-I antiviral pathway and is targeted by hepatitis C virus. Nature 437, 1167–1172.[CrossRef]
    [Google Scholar]
  18. Noguchi, M. & Hirohashi, S. ( 1996; ). Cell lines from non-neoplastic liver and hepatocellular carcinoma tissue from a single patient. In Vitro Cell Dev Biol Anim 32, 135–137.[CrossRef]
    [Google Scholar]
  19. Roy, C. R. & Mocarski, E. S. ( 2007; ). Pathogen subversion of cell-intrinsic innate immunity. Nat Immunol 8, 1179–1187.[CrossRef]
    [Google Scholar]
  20. Saito, T., Owen, D. M., Jiang, F., Marcotrigiano, J. & Gale, M., Jr ( 2008; ). Innate immunity induced by composition-dependent RIG-I recognition of hepatitis C virus RNA. Nature 454, 523–527.[CrossRef]
    [Google Scholar]
  21. Schröder, M. ( 2010; ). Human DEAD-box protein 3 has multiple functions in gene regulation and cell cycle control and is a prime target for viral manipulation. Biochem Pharmacol 79, 297–306.[CrossRef]
    [Google Scholar]
  22. Schröder, M., Baran, M. & Bowie, A. G. ( 2008; ). Viral targeting of DEAD box protein 3 reveals its role in TBK1/IKKϵ-mediated IRF activation. EMBO J 27, 2147–2157.[CrossRef]
    [Google Scholar]
  23. Seeger, C. & Mason, W. S. ( 2000; ). Hepatitis B virus biology. Microbiol Mol Biol Rev 64, 51–68.[CrossRef]
    [Google Scholar]
  24. Soulat, D., Burckstummer, T., Westermayer, S., Goncalves, A., Bauch, A., Stefanovic, A., Hantschel, O., Bennett, K. L., Decker, T. & Superti-Furga, G. ( 2008; ). The DEAD-box helicase DDX3X is a critical component of the TANK-binding kinase 1-dependent innate immune response. EMBO J 27, 2135–2146.[CrossRef]
    [Google Scholar]
  25. Takeuchi, O. & Akira, S. ( 2009; ). Innate immunity to virus infection. Immunol Rev 227, 75–86.[CrossRef]
    [Google Scholar]
  26. Taniguchi, T. & Takaoka, A. ( 2002; ). The interferon-α/β system in antiviral responses: a multimodal machinery of gene regulation by the IRF family of transcription factors. Curr Opin Immunol 14, 111–116.[CrossRef]
    [Google Scholar]
  27. van der Molen, R. G., Sprengers, D., Binda, R. S., de Jong, E. C., Niesters, H. G., Kusters, J. G., Kwekkeboom, J. & Janssen, H. L. ( 2004; ). Functional impairment of myeloid and plasmacytoid dendritic cells of patients with chronic hepatitis B. Hepatology 40, 738–746.[CrossRef]
    [Google Scholar]
  28. Wang, H. & Ryu, W.-S. ( 2009; ). HBV polymerase inhibits pattern recognition receptor-mediated innate immune response via its interaction with DDX3 DEAD-box RNA helicase. In Abstracts of The Molecular Biology of Hepatitis B Viruses meeting, Tours, 30 August–2 September 2009, p. 109. Doylestown, PA: Hepatitis B Foundation.
  29. Wang, H. & Ryu, W.-S. ( 2010; ). Hepatitis B virus polymerase blocks pattern recognition receptor signaling via interaction with DDX3: implications for immune evasion. PLoS Pathog 6, e1000986. doi:10.1371/journal.ppat.1000986
    [Google Scholar]
  30. Wang, H., Kim, S. & Ryu, W. S. ( 2009; ). DDX3 DEAD-Box RNA helicase inhibits hepatitis B virus reverse transcription by incorporation into nucleocapsids. J Virol 83, 5815–5824.[CrossRef]
    [Google Scholar]
  31. Wieland, S. F. & Chisari, F. V. ( 2005; ). Stealth and cunning: hepatitis B and hepatitis C viruses. J Virol 79, 9369–9380.[CrossRef]
    [Google Scholar]
  32. Wu, M., Xu, Y., Lin, S., Zhang, X., Xiang, L. & Yuan, Z. ( 2007; ). Hepatitis B virus polymerase inhibits the interferon-inducible MyD88 promoter by blocking nuclear translocation of Stat1. J Gen Virol 88, 3260–3269.[CrossRef]
    [Google Scholar]
  33. Wu, J., Meng, Z., Jiang, M., Pei, R., Trippler, M., Broering, R., Bucchi, A., Sowa, J. P., Dittmer, U. & other authors ( 2009; ). Hepatitis B virus suppresses Toll-like receptor-mediated innate immune responses in murine parenchymal and nonparenchymal liver cells. Hepatology 49, 1132–1140.[CrossRef]
    [Google Scholar]
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vol. , part 8, pp. 2080 - 2090

HBV polymerase inhibits IRF3 nuclear translocation

DDX3 is involved in inhibition of IFN-β induction by HBV polymerase [Single PDF file](199 KB)



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