1887

Abstract

Chronic viral hepatitis C is characterized by iron accumulation in the liver, and hepcidin regulates iron absorption. Hepatitis C virus (HCV) core+1/ARFP is a novel protein produced by a second functional ORF within the core gene. Here, using reporter assays and HCV bicistronic replicons, we show that, similarly to core, core+1/ARFP decreases hepcidin expression in hepatoma cells. The activator protein 1 (AP1) binding site of the human hepcidin promoter, shown here to be relevant to basal promoter activity and to the repression by core, is essential for the downregulation by core+1/ARFP while the previously described C/EBP (CCAAT/enhancer binding protein) and STAT (signal transducer and activator of transcription) sites are not. Consistently, expression of the AP1 components c-jun and c-fos obliterated the repressive effect of core and core+1/ARFP. In conclusion, we provide evidence that core+1/ARFP downregulates AP1-mediated transcription, providing new insights into the biological role of core+1/ARFP, as well as the transcriptional modulation of hepcidin, the main regulator of iron metabolism.

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2013-07-01
2020-07-14
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References

  1. Ahmad J., Eng F. J., Branch A. D. 2011; HCV and HCC: clinical update and a review of HCC-associated viral mutations in the core gene. Semin Liver Dis 31:347–355 [CrossRef][PubMed]
    [Google Scholar]
  2. Bakiri L., Matsuo K., Wisniewska M., Wagner E. F., Yaniv M. 2002; Promoter specificity and biological activity of tethered AP-1 dimers. Mol Cell Biol 22:4952–4964 [CrossRef][PubMed]
    [Google Scholar]
  3. Baril M., Brakier-Gingras L. 2005; Translation of the F protein of hepatitis C virus is initiated at a non-AUG codon in a +1 reading frame relative to the polyprotein. Nucleic Acids Res 33:1474–1486 [CrossRef][PubMed]
    [Google Scholar]
  4. Bartolomei G., Cevik R. E., Marcello A. 2011; Modulation of hepatitis C virus replication by iron and hepcidin in Huh7 hepatocytes. J Gen Virol 92:2072–2081 [CrossRef][PubMed]
    [Google Scholar]
  5. Boulant S., Becchi M., Penin F., Lavergne J. P. 2003; Unusual multiple recoding events leading to alternative forms of hepatitis C virus core protein from genotype 1b. J Biol Chem 278:45785–45792 [CrossRef][PubMed]
    [Google Scholar]
  6. Boumlic A., Nominé Y., Charbonnier S., Dalagiorgou G., Vassilaki N., Kieffer B., Travé G., Mavromara P., Orfanoudakis G. 2010; Prevalence of intrinsic disorder in the hepatitis C virus ARFP/Core+1/S protein. FEBS J 277:774–789 [CrossRef][PubMed]
    [Google Scholar]
  7. Boumlic A., Vassilaki N., Dalagiorgou G., Kochlios E., Kakkanas A., Georgopoulou U., Markoulatos P., Orfanoudakis G., Mavromara P. 2011; Internal translation initiation stimulates expression of the ARF/core+1 open reading frame of HCV genotype 1b. Virus Res 155:213–220 [CrossRef][PubMed]
    [Google Scholar]
  8. Branch A. D., Stump D. D., Gutierrez J. A., Eng F., Walewski J. L. 2005; The hepatitis C virus alternate reading frame (ARF) and its family of novel products: the alternate reading frame protein/F-protein, the double-frameshift protein, and others. Semin Liver Dis 25:105–117 [CrossRef][PubMed]
    [Google Scholar]
  9. Courselaud B., Pigeon C., Inoue Y., Inoue J., Gonzalez F. J., Leroyer P., Gilot D., Boudjema K., Guguen-Guillouzo C. other authors 2002; C/EBPalpha regulates hepatic transcription of hepcidin, an antimicrobial peptide and regulator of iron metabolism. Cross-talk between C/EBP pathway and iron metabolism. J Biol Chem 277:41163–41170 [CrossRef][PubMed]
    [Google Scholar]
  10. Dalagiorgou G., Vassilaki N., Foka P., Boumlic A., Kakkanas A., Kochlios E., Khalili S., Aslanoglou E., Veletza S. other authors 2011; High levels of HCV core+1 antibodies in HCV patients with hepatocellular carcinoma. J Gen Virol 92:1343–1351 [CrossRef][PubMed]
    [Google Scholar]
  11. Eisen-Vandervelde A. L., Waggoner S. N., Yao Z. Q., Cale E. M., Hahn C. S., Hahn Y. S. 2004; Hepatitis C virus core selectively suppresses interleukin-12 synthesis in human macrophages by interfering with AP-1 activation. J Biol Chem 279:43479–43486 [CrossRef][PubMed]
    [Google Scholar]
  12. 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 [CrossRef][PubMed]
    [Google Scholar]
  13. Erhardt A., Hassan M., Heintges T., Häussinger D. 2002; Hepatitis C virus core protein induces cell proliferation and activates ERK, JNK, and p38 MAP kinases together with the MAP kinase phosphatase MKP-1 in a HepG2 Tet-Off cell line. Virology 292:272–284 [CrossRef][PubMed]
    [Google Scholar]
  14. Fujita N., Sugimoto R., Takeo M., Urawa N., Mifuji R., Tanaka H., Kobayashi Y., Iwasa M., Watanabe S. other authors 2007; Hepcidin expression in the liver: relatively low level in patients with chronic hepatitis C. Mol Med 13:97–104 [CrossRef][PubMed]
    [Google Scholar]
  15. Hess J., Angel P., Schorpp-Kistner M. 2004; AP-1 subunits: quarrel and harmony among siblings. J Cell Sci 117:5965–5973 [CrossRef][PubMed]
    [Google Scholar]
  16. Hoofnagle J. H. 2002; Course and outcome of hepatitis C. Hepatology 36:Suppl 1S21–S29 [CrossRef][PubMed]
    [Google Scholar]
  17. Isoyama T., Kuge S., Nomoto A. 2002; The core protein of hepatitis C virus is imported into the nucleus by transport receptor Kap123p but inhibits Kap121p-dependent nuclear import of yeast AP1-like transcription factor in yeast cells. J Biol Chem 277:39634–39641 [CrossRef][PubMed]
    [Google Scholar]
  18. Jahan S., Ashfaq U. A., Khaliq S., Samreen B., Afzal N. 2012; Dual behavior of HCV Core gene in regulation of apoptosis is important in progression of HCC. Infect Genet Evol 12:236–239 [CrossRef][PubMed]
    [Google Scholar]
  19. Kim D. W., Suzuki R., Harada T., Saito I., Miyamura T. 1994; Trans-suppression of gene expression by hepatitis C viral core protein. Jpn J Med Sci Biol 47:211–220[PubMed] [CrossRef]
    [Google Scholar]
  20. Ko C., Siddaiah N., Berger J., Gish R., Brandhagen D., Sterling R. K., Cotler S. J., Fontana R. J., McCashland T. M. other authors 2007; Prevalence of hepatic iron overload and association with hepatocellular cancer in end-stage liver disease: results from the National Hemochromatosis Transplant Registry. Liver Int 27:1394–1401 [CrossRef][PubMed]
    [Google Scholar]
  21. Ma H. C., Lin T. W., Li H., Iguchi-Ariga S. M., Ariga H., Chuang Y. L., Ou J. H., Lo S. Y. 2008; Hepatitis C virus ARFP/F protein interacts with cellular MM-1 protein and enhances the gene trans-activation activity of c-Myc. J Biomed Sci 15:417–425 [CrossRef][PubMed]
    [Google Scholar]
  22. Mifuji R., Kobayashi Y., Ma N., Qiang Q. L., Urawa N., Horiike S., Iwasa M., Kaito M., Malavasi F., Adachi Y. 2006; Role of transferrin receptor 2 in hepatic accumulation of iron in patients with chronic hepatitis C. J Gastroenterol Hepatol 21:144–151 [CrossRef][PubMed]
    [Google Scholar]
  23. Miura K., Taura K., Kodama Y., Schnabl B., Brenner D. A. 2008; Hepatitis C virus-induced oxidative stress suppresses hepcidin expression through increased histone deacetylase activity. Hepatology 48:1420–1429 [CrossRef][PubMed]
    [Google Scholar]
  24. Nagashima M., Kudo M., Chung H., Ishikawa E., Hagiwara S., Nakatani T., Dote K. 2006; Regulatory failure of serum prohepcidin levels in patients with hepatitis C. Hepatol Res 36:288–293 [CrossRef][PubMed]
    [Google Scholar]
  25. Nemeth E., Ganz T. 2006; Regulation of iron metabolism by hepcidin. Annu Rev Nutr 26:323–342 [CrossRef][PubMed]
    [Google Scholar]
  26. Nishina S., Hino K., Korenaga M., Vecchi C., Pietrangelo A., Mizukami Y., Furutani T., Sakai A., Okuda M. other authors 2008; Hepatitis C virus-induced reactive oxygen species raise hepatic iron level in mice by reducing hepcidin transcription. Gastroenterology 134:226–238 [CrossRef][PubMed]
    [Google Scholar]
  27. Poenisch M., Bartenschlager R. 2010; New insights into structure and replication of the hepatitis C virus and clinical implications. Semin Liver Dis 30:333–347 [CrossRef][PubMed]
    [Google Scholar]
  28. Ratinier M., Boulant S., Crussard S., McLauchlan J., Lavergne J. P. 2009; Subcellular localizations of the hepatitis C virus alternate reading frame proteins. Virus Res 139:106–110 [CrossRef][PubMed]
    [Google Scholar]
  29. Ray R. B., Lagging L. M., Meyer K., Steele R., Ray R. 1995; Transcriptional regulation of cellular and viral promoters by the hepatitis C virus core protein. Virus Res 37:209–220 [CrossRef][PubMed]
    [Google Scholar]
  30. Roussel J., Pillez A., Montpellier C., Duverlie G., Cahour A., Dubuisson J., Wychowski C. 2003; Characterization of the expression of the hepatitis C virus F protein. J Gen Virol 84:1751–1759 [CrossRef][PubMed]
    [Google Scholar]
  31. Roy C. N., Mak H. H., Akpan I., Losyev G., Zurakowski D., Andrews N. C. 2007; Hepcidin antimicrobial peptide transgenic mice exhibit features of the anemia of inflammation. Blood 109:4038–4044 [CrossRef][PubMed]
    [Google Scholar]
  32. Ryan J. D., Altamura S., Devitt E., Mullins S., Lawless M. W., Muckenthaler M. U., Crowe J. 2012; Pegylated interferon-α induced hypoferremia is associated with the immediate response to treatment in hepatitis C. Hepatology 56:492–500 [CrossRef][PubMed]
    [Google Scholar]
  33. Shapiro V. S., Mollenauer M. N., Greene W. C., Weiss A. 1996; c-rel regulation of IL-2 gene expression may be mediated through activation of AP-1. J Exp Med 184:1663–1669 [CrossRef][PubMed]
    [Google Scholar]
  34. Shaulian E., Karin M. 2001; AP-1 in cell proliferation and survival. Oncogene 20:2390–2400 [CrossRef][PubMed]
    [Google Scholar]
  35. Sobesky R., Feray C., Rimlinger F., Derian N., Dos Santos A., Roque-Afonso A. M., Samuel D., Bréchot C., Thiers V. 2007; Distinct hepatitis C virus core and F protein quasispecies in tumoral and nontumoral hepatocytes isolated via microdissection. Hepatology 46:1704–1712 [CrossRef][PubMed]
    [Google Scholar]
  36. Tsutsumi T., Suzuki T., Moriya K., Yotsuyanagi H., Shintani Y., Fujie H., Matsuura Y., Kimura S., Koike K., Miyamura T. 2002; Alteration of intrahepatic cytokine expression and AP-1 activation in transgenic mice expressing hepatitis C virus core protein. Virology 304:415–424 [CrossRef][PubMed]
    [Google Scholar]
  37. Uversky V. N. 2011; Intrinsically disordered proteins from A to Z. Int J Biochem Cell Biol 43:1090–1103 [CrossRef][PubMed]
    [Google Scholar]
  38. Van Regenmortel M. H. V., Fauquet C. M., Bishop D. H. L., Carstens E. B., Estes M. K., Lemon S. M., Maniloff J., Mayo M. A., McGeoch D. J. other authors 2000 Virus Taxonomy: The VIIth Report of the International Committee on Taxonomy of Viruses San Diego, CA: Academic Press;
    [Google Scholar]
  39. Varaklioti A., Vassilaki N., Georgopoulou U., Mavromara P. 2002; Alternate translation occurs within the core coding region of the hepatitis C viral genome. J Biol Chem 277:17713–17721 [CrossRef][PubMed]
    [Google Scholar]
  40. Vassilaki N., Mavromara P. 2003; Two alternative translation mechanisms are responsible for the expression of the HCV ARFP/F/core+1 coding open reading frame. J Biol Chem 278:40503–40513 [CrossRef][PubMed]
    [Google Scholar]
  41. Vassilaki N., Mavromara P. 2009; The HCV ARFP/F/core+1 protein: production and functional analysis of an unconventional viral product. IUBMB Life 61:739–752 [CrossRef][PubMed]
    [Google Scholar]
  42. Vassilaki N., Boleti H., Mavromara P. 2007; Expression studies of the core+1 protein of the hepatitis C virus 1a in mammalian cells.. FEBS J 274:4057–4074 [CrossRef][PubMed]
    [Google Scholar]
  43. Vassilaki N., Boleti H., Mavromara P. 2008a; Expression studies of the HCV-1a core+1 open reading frame in mammalian cells. Virus Res 133:123–135 [CrossRef][PubMed]
    [Google Scholar]
  44. Vassilaki N., Friebe P., Meuleman P., Kallis S., Kaul A., Paranhos-Baccalà G., Leroux-Roels G., Mavromara P., Bartenschlager R. 2008b; 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 [CrossRef][PubMed]
    [Google Scholar]
  45. Verga Falzacappa M. V., Vujic Spasic M., Kessler R., Stolte J., Hentze M. W., Muckenthaler M. U. 2007; STAT3 mediates hepatic hepcidin expression and its inflammatory stimulation. Blood 109:353–358 [CrossRef][PubMed]
    [Google Scholar]
  46. Verga Falzacappa M. V., Casanovas G., Hentze M. W., Muckenthaler M. U. 2008; A bone morphogenetic protein (BMP)-responsive element in the hepcidin promoter controls HFE2-mediated hepatic hepcidin expression and its response to IL-6 in cultured cells. J Mol Med 86:531–540 [CrossRef][PubMed]
    [Google Scholar]
  47. Volke M., Gale D. P., Maegdefrau U., Schley G., Klanke B., Bosserhoff A. K., Maxwell P. H., Eckardt K. U., Warnecke C. 2009; Evidence for a lack of a direct transcriptional suppression of the iron regulatory peptide hepcidin by hypoxia-inducible factors. PLoS ONE 4:e7875 [CrossRef][PubMed]
    [Google Scholar]
  48. 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 [CrossRef][PubMed]
    [Google Scholar]
  49. Weizer-Stern O., Adamsky K., Margalit O., Ashur-Fabian O., Givol D., Amariglio N., Rechavi G. 2007; Hepcidin, a key regulator of iron metabolism, is transcriptionally activated by p53. Br J Haematol 138:253–262 [CrossRef][PubMed]
    [Google Scholar]
  50. 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 [CrossRef][PubMed]
    [Google Scholar]
  51. Xu Z., Choi J., Lu W., Ou J. H. 2003; Hepatitis C virus f protein is a short-lived protein associated with the endoplasmic reticulum. J Virol 77:1578–1583 [CrossRef][PubMed]
    [Google Scholar]
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