1887

Abstract

The human MxA protein is an interferon (IFN)-inducible GTPase with proven antiviral activity against diverse viruses. IFN responsiveness is impaired in chronic hepatitis B virus (HBV) infection. Accordingly, initial experiments showed that, in contrast to parental HepG2 cells, when HepG2-derived 2.2.15 liver cells carrying the HBV genome were treated with IFN, they could not synthesize the MxA protein. Furthermore, MxA expression was reduced in HepG2 cells transiently transfected with the HBV genome. To assess whether HBV-encoded precore/core (preC/C) proteins interact with the IFN-signalling pathway, HepG2, Chang and HeLa cells were transfected with preC/C expression plasmids; the levels of signal transducers remained unaffected. Next, full-length and deletion mutants fused to the CAT reporter gene were tested to investigate whether MxA inhibition occurs at the promoter level. In co-transfection experiments, IFN-induced CAT activity was inhibited by preC/C expression in a dose-dependent manner. Analysis of deletion mutants showed that the region affected by the preC/C proteins comprises IFN-stimulated response elements 2 and 3, upstream of the putative start codon of the MxA promoter. In addition, HBV preC/C proteins interacted directly with the MxA promoter, as shown by electrophoretic mobility shift assays. These results demonstrate a mechanism that HBV probably uses to downregulate an element of the IFN-induced host antiviral responses, which accounts for the impairment observed in HBV-infected patients.

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2003-08-01
2024-03-29
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References

  1. Aebi M., Fah J., Hurt N., Samuel C. E., Thomis D., Bazzigher L., Pavlovic J., Haller O., Staeheli P. 1989; cDNA structures and regulation of two interferon-induced human Mx proteins. Mol Cell Biol 9:5062–5072
    [Google Scholar]
  2. Aiba N., McGarvey M. J., Waters J., Hadziyannis S. J., Thomas H. C., Karayiannis P. 1997; The precore sequence of hepatitis B virus is required for nuclear localization of the core protein. Hepatology 26:1311–1317
    [Google Scholar]
  3. Brook M. G., Karayiannis P., Thomas H. C. 1989; Which patients with chronic hepatitis B virus infection will respond to alpha-interferon therapy? A statistical analysis of predictive factors. Hepatology 10:761–763
    [Google Scholar]
  4. Bruss V., Gerlich W. H. 1988; Formation of transmembraneous hepatitis B e-antigen by cotranslational in vitro processing of the viral precore protein. Virology 163:268–275
    [Google Scholar]
  5. Campillo M. L., Quiroga J. A., Bartolome J., Moraleda G., Castillo I., Carreño V. 1992; Protein composition of the hepatitis B virus e antigen in the natural course of disease and following interferon therapy. J Clin Microbiol 30:1256–1261
    [Google Scholar]
  6. Carman W. F., Jacyna M. R., Hadziyannis S., Karayiannis P., McGarvey M. J., Makris A., Thomas H. C. 1989; Mutation preventing formation of hepatitis B e antigen in patients with chronic hepatitis B infection. Lancet ii588–591
    [Google Scholar]
  7. Caselmann W. H. 1996; Trans -activation of cellular genes by hepatitis B virus proteins: a possible mechanism of hepatocarcinogenesis. Adv Virus Res 47:253–302
    [Google Scholar]
  8. Chang C., Enders G., Sprengel R., Peters N., Varmus H. E., Ganem D. 1987; Expression of the precore region of an avian hepatitis B virus is not required for viral replication. J Virol 61:3322–3325
    [Google Scholar]
  9. Chang K. C., Hansen E., Foroni L., Lida J., Goldspink G. 1991; Molecular and functional analysis of the virus- and interferon-inducible human MxA promoter. Arch Virol 117:1–15
    [Google Scholar]
  10. Fernandez M., Quiroga J. A., Martin J., Cotonat T., Pardo M., Horisberger M. A., Carreño V. 1997; Impaired interferon induction of human MxA protein in chronic hepatitis B virus infection. J Med Virol 51:332–337
    [Google Scholar]
  11. Frese M., Pietschmann T., Moradpour D., Haller O., Bartenschlager R. 2001; Interferon- α inhibits hepatitis C virus subgenomic RNA replication by an MxA-independent pathway. J Gen Virol 82:723–733
    [Google Scholar]
  12. Gordien E., Rosmorduc O., Peltekian C., Garreau F., Brechot C., Kremsdorf D. 2001; Inhibition of hepatitis B virus replication by the interferon-inducible MxA protein. J Virol 75:2684–2691
    [Google Scholar]
  13. Haque S. J., Williams B. R. 1998; Signal transduction in the interferon system. Semin Oncol 25:14–22
    [Google Scholar]
  14. Hatton T., Zhou S., Standring D. N. 1992; RNA- and DNA-binding activities in hepatitis B virus capsid protein: a model for their roles in viral replication. J Virol 66:5232–5241
    [Google Scholar]
  15. Heim M. H., Moradpour D., Blum H. E. 1999; Expression of hepatitis C virus proteins inhibits signal transduction through the Jak-STAT pathway. J Virol 73:8469–8475
    [Google Scholar]
  16. Henkler F., Hoare J., Waseem N., Goldin R. D., McGarvey M. J., Koshy R., King I. A. 2001; Intracellular localization of the hepatitis B virus HBx protein. J Gen Virol 82:871–882
    [Google Scholar]
  17. Horisberger M. A., McMaster G. K., Zeller H., Wathelet M. G., Dellis J., Content J. 1990; Cloning and sequence analyses of cDNAs for interferon- and virus-induced human Mx proteins reveal that they contain putative guanine nucleotide-binding sites: functional study of the corresponding gene promoter. J Virol 64:1171–1181
    [Google Scholar]
  18. Ihle J. N. 1996; STATs: signal transducers and activators of transcription. Cell 84:331–334
    [Google Scholar]
  19. Keskinen P., Nyqvist M., Sareneva T., Pirhonen J., Melen K., Julkunen I. 1999; Impaired antiviral response in human hepatoma cells. Virology 263:364–375
    [Google Scholar]
  20. Lee Y. H., Yun Y. 1998; HBx protein of hepatitis B virus activates Jak1-STAT signaling. J Biol Chem 273:25510–25515
    [Google Scholar]
  21. Lin O. S., Keeffe E. B. 2001; Current treatment strategies for chronic hepatitis B and C. Annu Rev Med 52:29–49
    [Google Scholar]
  22. Melen K., Keskinen P., Lehtonen A., Julkunen I. 2000; Interferon-induced gene expression and signaling in human hepatoma cell lines. J Hepatol 33:764–772
    [Google Scholar]
  23. Miller D. M., Zhang Y., Rahill B. M., Waldman W. J., Sedmak D. D. 1999; Human cytomegalovirus inhibits IFN- α -stimulated antiviral and immunoregulatory responses by blocking multiple levels of IFN- α signal transduction. J Immunol 162:6107–6113
    [Google Scholar]
  24. Muller U., Steinhoff U., Reis L. F., Hemmi S., Pavlovic J., Zinkernagel R. M., Aguet M. 1994; Functional role of type I and type II interferons in antiviral defense. Science 264:1918–1921
    [Google Scholar]
  25. Ono Y., Onda H., Sasada R., Igarashi K., Sugino Y., Nishioka K. 1983; The complete nucleotide sequences of the cloned hepatitis B virus DNA; subtype adr and adw . Nucleic Acids Res 11:1747–1757
    [Google Scholar]
  26. Pellegrini S., Dusanter-Fourt I. 1997; The structure, regulation and function of the Janus kinases (JAKs) and the signal transducers and activators of transcription (STATs). Eur J Biochem 248:615–633
    [Google Scholar]
  27. Quiroga J. A., Gonzalez O., Carreño V., Mora I., Porres J. C., Gutiez J., Hernandez-Guio C. 1985; Radioimmunoassay for detecting hepatitis B core antigen in serum from patients with chronic hepatitis B infection. Clin Chem 31:831–834
    [Google Scholar]
  28. Revel M., Chebath J. 1986; Interferon-activated genes. Trends Biochem Sci 11:166–170
    [Google Scholar]
  29. Ronni T., Matikainen S., Lehtonen A. 7 other authors 1998; The proximal interferon-stimulated response elements are essential for interferon responsiveness: a promoter analysis of the antiviral MxA gene. J Interferon Cytokine Res 18:773–781
    [Google Scholar]
  30. Rosmorduc O., Sirma H., Soussan P., Gordien E., Lebon P., Horisberger M., Brechot C., Kremsdorf D. 1999; Inhibition of interferon-inducible MxA protein expression by hepatitis B virus capsid protein. J Gen Virol 80:1253–1262
    [Google Scholar]
  31. Scaglioni P. P., Melegari M., Wands J. R. 1997; Posttranscriptional regulation of hepatitis B virus replication by the precore protein. J Virol 71:345–353
    [Google Scholar]
  32. Schlicht H. J., Salfeld J., Schaller H. 1987; The duck hepatitis B virus pre-C region encodes a signal sequence which is essential for synthesis and secretion of processed core proteins but not for virus formation. J Virol 61:3701–3709
    [Google Scholar]
  33. Sells M. A., Chen M. L., Acs G. 1987; Production of hepatitis B virus particles in Hep G2 cells transfected with cloned hepatitis B virus DNA. Proc Natl Acad Sci U S A 84:1005–1009
    [Google Scholar]
  34. Sen G. C., Ransohoff R. M. 1993; Interferon-induced antiviral actions and their regulation. Adv Virus Res 42:57–102
    [Google Scholar]
  35. Staeheli P. 1990; Interferon-induced proteins and the antiviral state. Adv Virus Res 38:147–200
    [Google Scholar]
  36. Stark G. R., Kerr I. M., Williams B. R., Silverman R. H., Schreiber R. D. 1998; How cells respond to interferons. Annu Rev Biochem 67:227–264
    [Google Scholar]
  37. Torresi J., Locarnini S. 2000; Antiviral chemotherapy for the treatment of hepatitis B virus infections. Gastroenterology 118 (Suppl. 1):S83–S103
    [Google Scholar]
  38. von Wussow P., Jakschies D., Hochkeppel H. K., Fibich C., Penner L., Deicher H. 1990; The human intracellular Mx-homologous protein is specifically induced by type I interferons. Eur J Immunol 20:2015–2019
    [Google Scholar]
  39. Whitten T. M., Quets A. T., Schloemer R. H. 1991; Identification of the hepatitis B virus factor that inhibits expression of the beta interferon gene. J Virol 65:4699–4704
    [Google Scholar]
  40. Williams B. R. 1991; Transcriptional regulation of interferon-stimulated genes. Eur J Biochem 200:1–11
    [Google Scholar]
  41. Zuckerman J. N., Zuckerman A. J. 2000; Current topics in hepatitis B. J Infect 41:130–136
    [Google Scholar]
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