Skip to main content
PMC home page
The Journal of Clinical Investigation logoLink to The Journal of Clinical Investigation
. 1996 Jan 15;97(2):388–395. doi: 10.1172/JCI118427

Regulation of transforming growth factor-beta 1 expression by the hepatitis B virus (HBV) X transactivator. Role in HBV pathogenesis.

Y D Yoo 1, H Ueda 1, K Park 1, K C Flanders 1, Y I Lee 1, G Jay 1, S J Kim 1
PMCID: PMC507029  PMID: 8567959

Abstract

TGF-beta 1 has been implicated in the pathogenesis of liver disease. The high frequency of detection of the hepatitis B virus X (HBx) antigen in liver cells from patients with chronic hepatitis, cirrhosis, and liver cancer suggested that expression of HBx and TGF-beta 1 may be associated. To test this possibility, we examined the expression of TGF-beta 1 in the liver of transgenic mice expressing the HBx gene. We show that the patterns of expression of TGF-beta 1 and Hbx protein are similar in these mice and that HBx activates transcription of the TGF-beta 1 gene in transfected hepatoma cells. The cis-acting element within the TGF-beta 1 gene that is responsive to regulation by Hbx is the binding site for the Egr family of transcription factors. We further show that the Egr-1 protein associates with the HBx protein, allowing HBx to participate in the transcriptional regulation of immediate-early genes. Our results suggest that expression of Hbx might induce expression of TGF-beta 1 in the early stages of infection and raise the possibility that TGF-beta 1 may play a role in hepatitis B virus pathogenesis.

Full Text

The Full Text of this article is available as a PDF (714.0 KB).

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Andus T., Bauer J., Gerok W. Effects of cytokines on the liver. Hepatology. 1991 Feb;13(2):364–375. [PubMed] [Google Scholar]
  2. Arii M., Takada S., Koike K. Identification of three essential regions of hepatitis B virus X protein for trans-activation function. Oncogene. 1992 Mar;7(3):397–403. [PubMed] [Google Scholar]
  3. Beasley R. P., Hwang L. Y., Lin C. C., Chien C. S. Hepatocellular carcinoma and hepatitis B virus. A prospective study of 22 707 men in Taiwan. Lancet. 1981 Nov 21;2(8256):1129–1133. doi: 10.1016/s0140-6736(81)90585-7. [DOI] [PubMed] [Google Scholar]
  4. Boyle W. J., Smeal T., Defize L. H., Angel P., Woodgett J. R., Karin M., Hunter T. Activation of protein kinase C decreases phosphorylation of c-Jun at sites that negatively regulate its DNA-binding activity. Cell. 1991 Feb 8;64(3):573–584. doi: 10.1016/0092-8674(91)90241-p. [DOI] [PubMed] [Google Scholar]
  5. Braun L., Mead J. E., Panzica M., Mikumo R., Bell G. I., Fausto N. Transforming growth factor beta mRNA increases during liver regeneration: a possible paracrine mechanism of growth regulation. Proc Natl Acad Sci U S A. 1988 Mar;85(5):1539–1543. doi: 10.1073/pnas.85.5.1539. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Cao X. M., Koski R. A., Gashler A., McKiernan M., Morris C. F., Gaffney R., Hay R. V., Sukhatme V. P. Identification and characterization of the Egr-1 gene product, a DNA-binding zinc finger protein induced by differentiation and growth signals. Mol Cell Biol. 1990 May;10(5):1931–1939. doi: 10.1128/mcb.10.5.1931. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Castilla A., Prieto J., Fausto N. Transforming growth factors beta 1 and alpha in chronic liver disease. Effects of interferon alfa therapy. N Engl J Med. 1991 Apr 4;324(14):933–940. doi: 10.1056/NEJM199104043241401. [DOI] [PubMed] [Google Scholar]
  8. Chen H. S., Kaneko S., Girones R., Anderson R. W., Hornbuckle W. E., Tennant B. C., Cote P. J., Gerin J. L., Purcell R. H., Miller R. H. The woodchuck hepatitis virus X gene is important for establishment of virus infection in woodchucks. J Virol. 1993 Mar;67(3):1218–1226. doi: 10.1128/jvi.67.3.1218-1226.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Christy B. A., Lau L. F., Nathans D. A gene activated in mouse 3T3 cells by serum growth factors encodes a protein with "zinc finger" sequences. Proc Natl Acad Sci U S A. 1988 Nov;85(21):7857–7861. doi: 10.1073/pnas.85.21.7857. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Christy B., Nathans D. DNA binding site of the growth factor-inducible protein Zif268. Proc Natl Acad Sci U S A. 1989 Nov;86(22):8737–8741. doi: 10.1073/pnas.86.22.8737. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Colgrove R., Simon G., Ganem D. Transcriptional activation of homologous and heterologous genes by the hepatitis B virus X gene product in cells permissive for viral replication. J Virol. 1989 Sep;63(9):4019–4026. doi: 10.1128/jvi.63.9.4019-4026.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Czaja M. J., Weiner F. R., Flanders K. C., Giambrone M. A., Wind R., Biempica L., Zern M. A. In vitro and in vivo association of transforming growth factor-beta 1 with hepatic fibrosis. J Cell Biol. 1989 Jun;108(6):2477–2482. doi: 10.1083/jcb.108.6.2477. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Derynck R., Goeddel D. V., Ullrich A., Gutterman J. U., Williams R. D., Bringman T. S., Berger W. H. Synthesis of messenger RNAs for transforming growth factors alpha and beta and the epidermal growth factor receptor by human tumors. Cancer Res. 1987 Feb 1;47(3):707–712. [PubMed] [Google Scholar]
  14. Drummond I. A., Madden S. L., Rohwer-Nutter P., Bell G. I., Sukhatme V. P., Rauscher F. J., 3rd Repression of the insulin-like growth factor II gene by the Wilms tumor suppressor WT1. Science. 1992 Jul 31;257(5070):674–678. doi: 10.1126/science.1323141. [DOI] [PubMed] [Google Scholar]
  15. Fausto N., Mead J. E. Regulation of liver growth: protooncogenes and transforming growth factors. Lab Invest. 1989 Jan;60(1):4–13. [PubMed] [Google Scholar]
  16. Flanders K. C., Thompson N. L., Cissel D. S., Van Obberghen-Schilling E., Baker C. C., Kass M. E., Ellingsworth L. R., Roberts A. B., Sporn M. B. Transforming growth factor-beta 1: histochemical localization with antibodies to different epitopes. J Cell Biol. 1989 Feb;108(2):653–660. doi: 10.1083/jcb.108.2.653. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Frankel S., Sohn R., Leinwand L. The use of sarkosyl in generating soluble protein after bacterial expression. Proc Natl Acad Sci U S A. 1991 Feb 15;88(4):1192–1196. doi: 10.1073/pnas.88.4.1192. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Gilardi P., Perricaudet M. The E4 transcriptional unit of Ad2: far upstream sequences are required for its transactivation by E1A. Nucleic Acids Res. 1984 Oct 25;12(20):7877–7888. doi: 10.1093/nar/12.20.7877. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Gonzalez G. A., Montminy M. R. Cyclic AMP stimulates somatostatin gene transcription by phosphorylation of CREB at serine 133. Cell. 1989 Nov 17;59(4):675–680. doi: 10.1016/0092-8674(89)90013-5. [DOI] [PubMed] [Google Scholar]
  20. Heine U., Munoz E. F., Flanders K. C., Ellingsworth L. R., Lam H. Y., Thompson N. L., Roberts A. B., Sporn M. B. Role of transforming growth factor-beta in the development of the mouse embryo. J Cell Biol. 1987 Dec;105(6 Pt 2):2861–2876. doi: 10.1083/jcb.105.6.2861. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Ito N., Kawata S., Tamura S., Takaishi K., Shirai Y., Kiso S., Yabuuchi I., Matsuda Y., Nishioka M., Tarui S. Elevated levels of transforming growth factor beta messenger RNA and its polypeptide in human hepatocellular carcinoma. Cancer Res. 1991 Aug 1;51(15):4080–4083. [PubMed] [Google Scholar]
  22. Kay A., Mandart E., Trepo C., Galibert F. The HBV HBX gene expressed in E. coli is recognised by sera from hepatitis patients. EMBO J. 1985 May;4(5):1287–1292. doi: 10.1002/j.1460-2075.1985.tb03774.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Kekulé A. S., Lauer U., Weiss L., Luber B., Hofschneider P. H. Hepatitis B virus transactivator HBx uses a tumour promoter signalling pathway. Nature. 1993 Feb 25;361(6414):742–745. doi: 10.1038/361742a0. [DOI] [PubMed] [Google Scholar]
  24. Kim C. M., Koike K., Saito I., Miyamura T., Jay G. HBx gene of hepatitis B virus induces liver cancer in transgenic mice. Nature. 1991 May 23;351(6324):317–320. doi: 10.1038/351317a0. [DOI] [PubMed] [Google Scholar]
  25. Kim S. J., Glick A., Sporn M. B., Roberts A. B. Characterization of the promoter region of the human transforming growth factor-beta 1 gene. J Biol Chem. 1989 Jan 5;264(1):402–408. [PubMed] [Google Scholar]
  26. Kim S. J., Kehrl J. H., Burton J., Tendler C. L., Jeang K. T., Danielpour D., Thevenin C., Kim K. Y., Sporn M. B., Roberts A. B. Transactivation of the transforming growth factor beta 1 (TGF-beta 1) gene by human T lymphotropic virus type 1 tax: a potential mechanism for the increased production of TGF-beta 1 in adult T cell leukemia. J Exp Med. 1990 Jul 1;172(1):121–129. doi: 10.1084/jem.172.1.121. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Kim S. J., Park K., Rudkin B. B., Dey B. R., Sporn M. B., Roberts A. B. Nerve growth factor induces transcription of transforming growth factor-beta 1 through a specific promoter element in PC12 cells. J Biol Chem. 1994 Feb 4;269(5):3739–3744. [PubMed] [Google Scholar]
  28. Kim S. J., Wagner S., Liu F., O'Reilly M. A., Robbins P. D., Green M. R. Retinoblastoma gene product activates expression of the human TGF-beta 2 gene through transcription factor ATF-2. Nature. 1992 Jul 23;358(6384):331–334. doi: 10.1038/358331a0. [DOI] [PubMed] [Google Scholar]
  29. Kwee L., Lucito R., Aufiero B., Schneider R. J. Alternate translation initiation on hepatitis B virus X mRNA produces multiple polypeptides that differentially transactivate class II and III promoters. J Virol. 1992 Jul;66(7):4382–4389. doi: 10.1128/jvi.66.7.4382-4389.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Lau L. F., Nathans D. Expression of a set of growth-related immediate early genes in BALB/c 3T3 cells: coordinate regulation with c-fos or c-myc. Proc Natl Acad Sci U S A. 1987 Mar;84(5):1182–1186. doi: 10.1073/pnas.84.5.1182. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Lemaire P., Revelant O., Bravo R., Charnay P. Two mouse genes encoding potential transcription factors with identical DNA-binding domains are activated by growth factors in cultured cells. Proc Natl Acad Sci U S A. 1988 Jul;85(13):4691–4695. doi: 10.1073/pnas.85.13.4691. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Levrero M., Balsano C., Natoli G., Avantaggiati M. L., Elfassi E. Hepatitis B virus X protein transactivates the long terminal repeats of human immunodeficiency virus types 1 and 2. J Virol. 1990 Jun;64(6):3082–3086. doi: 10.1128/jvi.64.6.3082-3086.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Lillie J. W., Green M. R. Transcription activation by the adenovirus E1a protein. Nature. 1989 Mar 2;338(6210):39–44. doi: 10.1038/338039a0. [DOI] [PubMed] [Google Scholar]
  34. Lim R. W., Varnum B. C., Herschman H. R. Cloning of tetradecanoyl phorbol ester-induced 'primary response' sequences and their expression in density-arrested Swiss 3T3 cells and a TPA non-proliferative variant. Oncogene. 1987;1(3):263–270. [PubMed] [Google Scholar]
  35. Liu F., Green M. R. A specific member of the ATF transcription factor family can mediate transcription activation by the adenovirus E1a protein. Cell. 1990 Jun 29;61(7):1217–1224. doi: 10.1016/0092-8674(90)90686-9. [DOI] [PubMed] [Google Scholar]
  36. Madden S. L., Cook D. M., Morris J. F., Gashler A., Sukhatme V. P., Rauscher F. J., 3rd Transcriptional repression mediated by the WT1 Wilms tumor gene product. Science. 1991 Sep 27;253(5027):1550–1553. doi: 10.1126/science.1654597. [DOI] [PubMed] [Google Scholar]
  37. Maguire H. F., Hoeffler J. P., Siddiqui A. HBV X protein alters the DNA binding specificity of CREB and ATF-2 by protein-protein interactions. Science. 1991 May 10;252(5007):842–844. doi: 10.1126/science.1827531. [DOI] [PubMed] [Google Scholar]
  38. Milbrandt J. A nerve growth factor-induced gene encodes a possible transcriptional regulatory factor. Science. 1987 Nov 6;238(4828):797–799. doi: 10.1126/science.3672127. [DOI] [PubMed] [Google Scholar]
  39. Moriarty A. M., Alexander H., Lerner R. A., Thornton G. B. Antibodies to peptides detect new hepatitis B antigen: serological correlation with hepatocellular carcinoma. Science. 1985 Jan 25;227(4685):429–433. doi: 10.1126/science.2981434. [DOI] [PubMed] [Google Scholar]
  40. Nakamura T., Tomita Y., Hirai R., Yamaoka K., Kaji K., Ichihara A. Inhibitory effect of transforming growth factor-beta on DNA synthesis of adult rat hepatocytes in primary culture. Biochem Biophys Res Commun. 1985 Dec 31;133(3):1042–1050. doi: 10.1016/0006-291x(85)91241-0. [DOI] [PubMed] [Google Scholar]
  41. Nakatsukasa H., Nagy P., Evarts R. P., Hsia C. C., Marsden E., Thorgeirsson S. S. Cellular distribution of transforming growth factor-beta 1 and procollagen types I, III, and IV transcripts in carbon tetrachloride-induced rat liver fibrosis. J Clin Invest. 1990 Jun;85(6):1833–1843. doi: 10.1172/JCI114643. [DOI] [PMC free article] [PubMed] [Google Scholar]
  42. Natoli G., Avantaggiati M. L., Chirillo P., Costanzo A., Artini M., Balsano C., Levrero M. Induction of the DNA-binding activity of c-jun/c-fos heterodimers by the hepatitis B virus transactivator pX. Mol Cell Biol. 1994 Feb;14(2):989–998. doi: 10.1128/mcb.14.2.989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  43. Panduro A., Shalaby F., Biempica L., Shafritz D. A. Changes in albumin, alpha-fetoprotein and collagen gene transcription in CCl4-induced hepatic fibrosis. Hepatology. 1988 Mar-Apr;8(2):259–266. doi: 10.1002/hep.1840080212. [DOI] [PubMed] [Google Scholar]
  44. Rauscher F. J., 3rd, Morris J. F., Tournay O. E., Cook D. M., Curran T. Binding of the Wilms' tumor locus zinc finger protein to the EGR-1 consensus sequence. Science. 1990 Nov 30;250(4985):1259–1262. doi: 10.1126/science.2244209. [DOI] [PubMed] [Google Scholar]
  45. Ritter S. E., Whitten T. M., Quets A. T., Schloemer R. H. An internal domain of the hepatitis B virus X antigen is necessary for transactivating activity. Virology. 1991 Jun;182(2):841–845. doi: 10.1016/0042-6822(91)90626-m. [DOI] [PubMed] [Google Scholar]
  46. Sadowski I., Ma J., Triezenberg S., Ptashne M. GAL4-VP16 is an unusually potent transcriptional activator. Nature. 1988 Oct 6;335(6190):563–564. doi: 10.1038/335563a0. [DOI] [PubMed] [Google Scholar]
  47. Seto E., Mitchell P. J., Yen T. S. Transactivation by the hepatitis B virus X protein depends on AP-2 and other transcription factors. Nature. 1990 Mar 1;344(6261):72–74. doi: 10.1038/344072a0. [DOI] [PubMed] [Google Scholar]
  48. Seto E., Zhou D. X., Peterlin B. M., Yen T. S. trans-activation by the hepatitis B virus X protein shows cell-type specificity. Virology. 1989 Dec;173(2):764–766. doi: 10.1016/0042-6822(89)90594-1. [DOI] [PubMed] [Google Scholar]
  49. Sheng M., Thompson M. A., Greenberg M. E. CREB: a Ca(2+)-regulated transcription factor phosphorylated by calmodulin-dependent kinases. Science. 1991 Jun 7;252(5011):1427–1430. doi: 10.1126/science.1646483. [DOI] [PubMed] [Google Scholar]
  50. Siddiqui A., Gaynor R., Srinivasan A., Mapoles J., Farr R. W. trans-activation of viral enhancers including long terminal repeat of the human immunodeficiency virus by the hepatitis B virus X protein. Virology. 1989 Apr;169(2):479–484. doi: 10.1016/0042-6822(89)90177-3. [DOI] [PubMed] [Google Scholar]
  51. Siddiqui A., Jameel S., Mapoles J. Expression of the hepatitis B virus X gene in mammalian cells. Proc Natl Acad Sci U S A. 1987 Apr;84(8):2513–2517. doi: 10.1073/pnas.84.8.2513. [DOI] [PMC free article] [PubMed] [Google Scholar]
  52. Sporn M. B., Roberts A. B. Autocrine growth factors and cancer. 1985 Feb 28-Mar 6Nature. 313(6005):745–747. doi: 10.1038/313745a0. [DOI] [PubMed] [Google Scholar]
  53. Sukhatme V. P. Early transcriptional events in cell growth: the Egr family. J Am Soc Nephrol. 1990 Dec;1(6):859–866. doi: 10.1681/ASN.V16859. [DOI] [PubMed] [Google Scholar]
  54. Takada S., Koike K. Trans-activation function of a 3' truncated X gene-cell fusion product from integrated hepatitis B virus DNA in chronic hepatitis tissues. Proc Natl Acad Sci U S A. 1990 Aug;87(15):5628–5632. doi: 10.1073/pnas.87.15.5628. [DOI] [PMC free article] [PubMed] [Google Scholar]
  55. Tiollais P., Pourcel C., Dejean A. The hepatitis B virus. Nature. 1985 Oct 10;317(6037):489–495. doi: 10.1038/317489a0. [DOI] [PubMed] [Google Scholar]
  56. Tratner I., Ofir R., Verma I. M. Alteration of a cyclic AMP-dependent protein kinase phosphorylation site in the c-Fos protein augments its transforming potential. Mol Cell Biol. 1992 Mar;12(3):998–1006. doi: 10.1128/mcb.12.3.998. [DOI] [PMC free article] [PubMed] [Google Scholar]
  57. Twu J. S., Lai M. Y., Chen D. S., Robinson W. S. Activation of protooncogene c-jun by the X protein of hepatitis B virus. Virology. 1993 Jan;192(1):346–350. doi: 10.1006/viro.1993.1041. [DOI] [PubMed] [Google Scholar]
  58. Twu J. S., Robinson W. S. Hepatitis B virus X gene can transactivate heterologous viral sequences. Proc Natl Acad Sci U S A. 1989 Mar;86(6):2046–2050. doi: 10.1073/pnas.86.6.2046. [DOI] [PMC free article] [PubMed] [Google Scholar]
  59. Twu J. S., Schloemer R. H. Transcriptional trans-activating function of hepatitis B virus. J Virol. 1987 Nov;61(11):3448–3453. doi: 10.1128/jvi.61.11.3448-3453.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  60. Ueda H., Ullrich S. J., Gangemi J. D., Kappel C. A., Ngo L., Feitelson M. A., Jay G. Functional inactivation but not structural mutation of p53 causes liver cancer. Nat Genet. 1995 Jan;9(1):41–47. doi: 10.1038/ng0195-41. [DOI] [PubMed] [Google Scholar]
  61. Unger T., Shaul Y. The X protein of the hepatitis B virus acts as a transcription factor when targeted to its responsive element. EMBO J. 1990 Jun;9(6):1889–1895. doi: 10.1002/j.1460-2075.1990.tb08315.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  62. Unsal H., Yakicier C., Marçais C., Kew M., Volkmann M., Zentgraf H., Isselbacher K. J., Ozturk M. Genetic heterogeneity of hepatocellular carcinoma. Proc Natl Acad Sci U S A. 1994 Jan 18;91(2):822–826. doi: 10.1073/pnas.91.2.822. [DOI] [PMC free article] [PubMed] [Google Scholar]
  63. Wang W. L., London W. T., Feitelson M. A. Hepatitis B x antigen in hepatitis B virus carrier patients with liver cancer. Cancer Res. 1991 Sep 15;51(18):4971–4977. [PubMed] [Google Scholar]
  64. Wang W. L., London W. T., Lega L., Feitelson M. A. HBxAg in the liver from carrier patients with chronic hepatitis and cirrhosis. Hepatology. 1991 Jul;14(1):29–37. doi: 10.1002/hep.1840140106. [DOI] [PubMed] [Google Scholar]
  65. Wang X. W., Forrester K., Yeh H., Feitelson M. A., Gu J. R., Harris C. C. Hepatitis B virus X protein inhibits p53 sequence-specific DNA binding, transcriptional activity, and association with transcription factor ERCC3. Proc Natl Acad Sci U S A. 1994 Mar 15;91(6):2230–2234. doi: 10.1073/pnas.91.6.2230. [DOI] [PMC free article] [PubMed] [Google Scholar]
  66. Yamamoto K. K., Gonzalez G. A., Biggs W. H., 3rd, Montminy M. R. Phosphorylation-induced binding and transcriptional efficacy of nuclear factor CREB. Nature. 1988 Aug 11;334(6182):494–498. doi: 10.1038/334494a0. [DOI] [PubMed] [Google Scholar]

Articles from Journal of Clinical Investigation are provided here courtesy of American Society for Clinical Investigation

RESOURCES