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. 1990 May;64(5):2360–2368. doi: 10.1128/jvi.64.5.2360-2368.1990

Differentiation-specific transcriptional regulation of the hepatitis B virus large surface antigen gene in human hepatoma cell lines.

A K Raney 1, D R Milich 1, A J Easton 1, A McLachlan 1
PMCID: PMC249397  PMID: 2157890

Abstract

The transcriptional activities of the four hepatitis B virus promoters were compared in three differentiated hepatoma cell lines, HepG2, Hep3B, and PLC/PRF/5; a dedifferentiated subline of HepG2, HepG2.1; a human cervical carcinoma cell line, HeLa S3; and a mouse fibroblast cell line, NIH 3T3. The plasmid constructs, which contain the complete hepatitis B virus genome directing the expression of the luciferase reporter gene, were analyzed by transient transfection assays. The relative orders of the levels of the transcriptional activities of the four promoters were similar in each of the cell lines. The major surface antigen and X-gene promoters displayed the highest activity levels, the core promoter activity level was less than or similar to the activity levels of these two promoters, and the large surface antigen promoter had the lowest activity level in all of the cell lines examined. The core promoter demonstrated an approximately 2- to 20-fold higher relative level of expression in the differentiated hepatoma cell lines, suggesting that this promoter might be preferentially active in these cells. The relative level of activity of the large surface antigen promoter in the differentiated hepatoma cell lines was approximately 5 to 90 times greater than that observed in the other cell lines, indicating that the activity of this promoter is highly specific for differentiation state and cell type. Deletion analysis of the large surface antigen promoter demonstrated that the sequence element responsible for the differentiation state-specific expression from this promoter is located between nucleotides 2719 and 2733 (-90 and -76). Within this sequence element is a binding site (GTTAATCATTACT) for the liver-specific transcription factor hepatocyte nuclear factor 1 (HNF1). This indicates that the preferential expression from the large surface antigen promoter in the differentiated hepatoma cell lines is probably mediated by HNF1 or an HNF1-related transcription factor.

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Selected References

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  1. Antonucci T. K., Rutter W. J. Hepatitis B virus (HBV) promoters are regulated by the HBV enhancer in a tissue-specific manner. J Virol. 1989 Feb;63(2):579–583. doi: 10.1128/jvi.63.2.579-583.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Araki K., Miyazaki J., Hino O., Tomita N., Chisaka O., Matsubara K., Yamamura K. Expression and replication of hepatitis B virus genome in transgenic mice. Proc Natl Acad Sci U S A. 1989 Jan;86(1):207–211. doi: 10.1073/pnas.86.1.207. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Baumhueter S., Courtois G., Crabtree G. R. A variant nuclear protein in dedifferentiated hepatoma cells binds to the same functional sequences in the beta fibrinogen gene promoter as HNF-1. EMBO J. 1988 Aug;7(8):2485–2493. doi: 10.1002/j.1460-2075.1988.tb03095.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Bavand M., Feitelson M., Laub O. The hepatitis B virus-associated reverse transcriptase is encoded by the viral pol gene. J Virol. 1989 Feb;63(2):1019–1021. doi: 10.1128/jvi.63.2.1019-1021.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Cattaneo R., Will H., Hernandez N., Schaller H. Signals regulating hepatitis B surface antigen transcription. Nature. 1983 Sep 22;305(5932):336–338. doi: 10.1038/305336a0. [DOI] [PubMed] [Google Scholar]
  6. Cattaneo R., Will H., Schaller H. Hepatitis B virus transcription in the infected liver. EMBO J. 1984 Sep;3(9):2191–2196. doi: 10.1002/j.1460-2075.1984.tb02113.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Chang C. M., Jeng K. S., Hu C. P., Lo S. J., Su T. S., Ting L. P., Chou C. K., Han S. H., Pfaff E., Salfeld J. Production of hepatitis B virus in vitro by transient expression of cloned HBV DNA in a hepatoma cell line. EMBO J. 1987 Mar;6(3):675–680. doi: 10.1002/j.1460-2075.1987.tb04807.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Chang H. K., Wang B. Y., Yuh C. H., Wei C. L., Ting L. P. A liver-specific nuclear factor interacts with the promoter region of the large surface protein gene of human hepatitis B virus. Mol Cell Biol. 1989 Nov;9(11):5189–5197. doi: 10.1128/mcb.9.11.5189. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Chang L. J., Pryciak P., Ganem D., Varmus H. E. Biosynthesis of the reverse transcriptase of hepatitis B viruses involves de novo translational initiation not ribosomal frameshifting. Nature. 1989 Jan 26;337(6205):364–368. doi: 10.1038/337364a0. [DOI] [PubMed] [Google Scholar]
  10. Cheng K. C., Smith G. L., Moss B. Hepatitis B virus large surface protein is not secreted but is immunogenic when selectively expressed by recombinant vaccinia virus. J Virol. 1986 Nov;60(2):337–344. doi: 10.1128/jvi.60.2.337-344.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Chisari F. V., Filippi P., McLachlan A., Milich D. R., Riggs M., Lee S., Palmiter R. D., Pinkert C. A., Brinster R. L. Expression of hepatitis B virus large envelope polypeptide inhibits hepatitis B surface antigen secretion in transgenic mice. J Virol. 1986 Dec;60(3):880–887. doi: 10.1128/jvi.60.3.880-887.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. 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]
  13. Courtois G., Baumhueter S., Crabtree G. R. Purified hepatocyte nuclear factor 1 interacts with a family of hepatocyte-specific promoters. Proc Natl Acad Sci U S A. 1988 Nov;85(21):7937–7941. doi: 10.1073/pnas.85.21.7937. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Courtois G., Morgan J. G., Campbell L. A., Fourel G., Crabtree G. R. Interaction of a liver-specific nuclear factor with the fibrinogen and alpha 1-antitrypsin promoters. Science. 1987 Oct 30;238(4827):688–692. doi: 10.1126/science.3499668. [DOI] [PubMed] [Google Scholar]
  15. De-Medina T., Faktor O., Shaul Y. The S promoter of hepatitis B virus is regulated by positive and negative elements. Mol Cell Biol. 1988 Jun;8(6):2449–2455. doi: 10.1128/mcb.8.6.2449. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Dubois M. F., Pourcel C., Rousset S., Chany C., Tiollais P. Excretion of hepatitis B surface antigen particles from mouse cells transformed with cloned viral DNA. Proc Natl Acad Sci U S A. 1980 Aug;77(8):4549–4553. doi: 10.1073/pnas.77.8.4549. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Farza H., Hadchouel M., Scotto J., Tiollais P., Babinet C., Pourcel C. Replication and gene expression of hepatitis B virus in a transgenic mouse that contains the complete viral genome. J Virol. 1988 Nov;62(11):4144–4152. doi: 10.1128/jvi.62.11.4144-4152.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Frain M., Swart G., Monaci P., Nicosia A., Stämpfli S., Frank R., Cortese R. The liver-specific transcription factor LF-B1 contains a highly diverged homeobox DNA binding domain. Cell. 1989 Oct 6;59(1):145–157. doi: 10.1016/0092-8674(89)90877-5. [DOI] [PubMed] [Google Scholar]
  19. Ganem D., Varmus H. E. The molecular biology of the hepatitis B viruses. Annu Rev Biochem. 1987;56:651–693. doi: 10.1146/annurev.bi.56.070187.003251. [DOI] [PubMed] [Google Scholar]
  20. Gorman C. M., Moffat L. F., Howard B. H. Recombinant genomes which express chloramphenicol acetyltransferase in mammalian cells. Mol Cell Biol. 1982 Sep;2(9):1044–1051. doi: 10.1128/mcb.2.9.1044. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Gripon P., Diot C., Thézé N., Fourel I., Loreal O., Brechot C., Guguen-Guillouzo C. Hepatitis B virus infection of adult human hepatocytes cultured in the presence of dimethyl sulfoxide. J Virol. 1988 Nov;62(11):4136–4143. doi: 10.1128/jvi.62.11.4136-4143.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Hardon E. M., Frain M., Paonessa G., Cortese R. Two distinct factors interact with the promoter regions of several liver-specific genes. EMBO J. 1988 Jun;7(6):1711–1719. doi: 10.1002/j.1460-2075.1988.tb03000.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Heermann K. H., Goldmann U., Schwartz W., Seyffarth T., Baumgarten H., Gerlich W. H. Large surface proteins of hepatitis B virus containing the pre-s sequence. J Virol. 1984 Nov;52(2):396–402. doi: 10.1128/jvi.52.2.396-402.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Honigwachs J., Faktor O., Dikstein R., Shaul Y., Laub O. Liver-specific expression of hepatitis B virus is determined by the combined action of the core gene promoter and the enhancer. J Virol. 1989 Feb;63(2):919–924. doi: 10.1128/jvi.63.2.919-924.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Jameel S., Siddiqui A. The human hepatitis B virus enhancer requires trans-acting cellular factor(s) for activity. Mol Cell Biol. 1986 Feb;6(2):710–715. doi: 10.1128/mcb.6.2.710. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Junker M., Galle P., Schaller H. Expression and replication of the hepatitis B virus genome under foreign promoter control. Nucleic Acids Res. 1987 Dec 23;15(24):10117–10132. doi: 10.1093/nar/15.24.10117. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Kaneko S., Miller R. H. X-region-specific transcript in mammalian hepatitis B virus-infected liver. J Virol. 1988 Nov;62(11):3979–3984. doi: 10.1128/jvi.62.11.3979-3984.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Karpen S., Banerjee R., Zelent A., Price P., Acs G. Identification of protein-binding sites in the hepatitis B virus enhancer and core promoter domains. Mol Cell Biol. 1988 Dec;8(12):5159–5165. doi: 10.1128/mcb.8.12.5159. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Lichtsteiner S., Schibler U. A glycosylated liver-specific transcription factor stimulates transcription of the albumin gene. Cell. 1989 Jun 30;57(7):1179–1187. doi: 10.1016/0092-8674(89)90055-x. [DOI] [PubMed] [Google Scholar]
  30. Maire P., Wuarin J., Schibler U. The role of cis-acting promoter elements in tissue-specific albumin gene expression. Science. 1989 Apr 21;244(4902):343–346. doi: 10.1126/science.2711183. [DOI] [PubMed] [Google Scholar]
  31. McLachlan A., Milich D. R., Raney A. K., Riggs M. G., Hughes J. L., Sorge J., Chisari F. V. Expression of hepatitis B virus surface and core antigens: influences of pre-S and precore sequences. J Virol. 1987 Mar;61(3):683–692. doi: 10.1128/jvi.61.3.683-692.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Ou J. H., Rutter W. J. Regulation of secretion of the hepatitis B virus major surface antigen by the preS-1 protein. J Virol. 1987 Mar;61(3):782–786. doi: 10.1128/jvi.61.3.782-786.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Persing D. H., Varmus H. E., Ganem D. A frameshift mutation in the pre-S region of the human hepatitis B virus genome allows production of surface antigen particles but eliminates binding to polymerized albumin. Proc Natl Acad Sci U S A. 1985 May;82(10):3440–3444. doi: 10.1073/pnas.82.10.3440. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Persing D. H., Varmus H. E., Ganem D. Inhibition of secretion of hepatitis B surface antigen by a related presurface polypeptide. Science. 1986 Dec 12;234(4782):1388–1391. doi: 10.1126/science.3787251. [DOI] [PubMed] [Google Scholar]
  35. Pourcel C., Louise A., Gervais M., Chenciner N., Dubois M. F., Tiollais P. Transcription of the hepatitis B surface antigen gene in mouse cells transformed with cloned viral DNA. J Virol. 1982 Apr;42(1):100–105. doi: 10.1128/jvi.42.1.100-105.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Raney A. K., Milich D. R., McLachlan A. Characterization of hepatitis B virus major surface antigen gene transcriptional regulatory elements in differentiated hepatoma cell lines. J Virol. 1989 Sep;63(9):3919–3925. doi: 10.1128/jvi.63.9.3919-3925.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. Richards W. L., Tsukada Y., Potter V. R. Phenotypic diversity of gamma-glutamyltranspeptidase activity and protein secretion in hepatoma cell lines. Cancer Res. 1982 Apr;42(4):1374–1383. [PubMed] [Google Scholar]
  38. Rijntjes P. J., Moshage H. J., Yap S. H. In vitro infection of primary cultures of cryopreserved adult human hepatocytes with hepatitis B virus. Virus Res. 1988 Apr;10(1):95–109. doi: 10.1016/0168-1702(88)90060-3. [DOI] [PubMed] [Google Scholar]
  39. Sanger F., Nicklen S., Coulson A. R. DNA sequencing with chain-terminating inhibitors. Proc Natl Acad Sci U S A. 1977 Dec;74(12):5463–5467. doi: 10.1073/pnas.74.12.5463. [DOI] [PMC free article] [PubMed] [Google Scholar]
  40. Sawadaishi K., Morinaga T., Tamaoki T. Interaction of a hepatoma-specific nuclear factor with transcription-regulatory sequences of the human alpha-fetoprotein and albumin genes. Mol Cell Biol. 1988 Dec;8(12):5179–5187. doi: 10.1128/mcb.8.12.5179. [DOI] [PMC free article] [PubMed] [Google Scholar]
  41. Seeger C., Baldwin B., Tennant B. C. Expression of infectious woodchuck hepatitis virus in murine and avian fibroblasts. J Virol. 1989 Nov;63(11):4665–4669. doi: 10.1128/jvi.63.11.4665-4669.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  42. Sells M. A., Chen M. L., Acs G. Production of hepatitis B virus particles in Hep G2 cells transfected with cloned hepatitis B virus DNA. Proc Natl Acad Sci U S A. 1987 Feb;84(4):1005–1009. doi: 10.1073/pnas.84.4.1005. [DOI] [PMC free article] [PubMed] [Google Scholar]
  43. Sells M. A., Zelent A. Z., Shvartsman M., Acs G. Replicative intermediates of hepatitis B virus in HepG2 cells that produce infectious virions. J Virol. 1988 Aug;62(8):2836–2844. doi: 10.1128/jvi.62.8.2836-2844.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  44. Seto E., Yen T. S., Peterlin B. M., Ou J. H. Trans-activation of the human immunodeficiency virus long terminal repeat by the hepatitis B virus X protein. Proc Natl Acad Sci U S A. 1988 Nov;85(21):8286–8290. doi: 10.1073/pnas.85.21.8286. [DOI] [PMC free article] [PubMed] [Google Scholar]
  45. Shih C. H., Li L. S., Roychoudhury S., Ho M. H. In vitro propagation of human hepatitis B virus in a rat hepatoma cell line. Proc Natl Acad Sci U S A. 1989 Aug;86(16):6323–6327. doi: 10.1073/pnas.86.16.6323. [DOI] [PMC free article] [PubMed] [Google Scholar]
  46. 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]
  47. 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]
  48. Siddiqui A., Jameel S., Mapoles J. Transcriptional control elements of hepatitis B surface antigen gene. Proc Natl Acad Sci U S A. 1986 Feb;83(3):566–570. doi: 10.1073/pnas.83.3.566. [DOI] [PMC free article] [PubMed] [Google Scholar]
  49. Spandau D. F., Lee C. H. trans-activation of viral enhancers by the hepatitis B virus X protein. J Virol. 1988 Feb;62(2):427–434. doi: 10.1128/jvi.62.2.427-434.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  50. Standring D. N., Ou J. H., Rutter W. J. Assembly of viral particles in Xenopus oocytes: pre-surface-antigens regulate secretion of the hepatitis B viral surface envelope particle. Proc Natl Acad Sci U S A. 1986 Dec;83(24):9338–9342. doi: 10.1073/pnas.83.24.9338. [DOI] [PMC free article] [PubMed] [Google Scholar]
  51. Su T. S., Lui W. Y., Lin L. H., Han S. H., P'eng F. K. Analysis of hepatitis B virus transcripts in infected human livers. Hepatology. 1989 Feb;9(2):180–185. doi: 10.1002/hep.1840090203. [DOI] [PubMed] [Google Scholar]
  52. Sureau C., Romet-Lemonne J. L., Mullins J. I., Essex M. Production of hepatitis B virus by a differentiated human hepatoma cell line after transfection with cloned circular HBV DNA. Cell. 1986 Oct 10;47(1):37–47. doi: 10.1016/0092-8674(86)90364-8. [DOI] [PubMed] [Google Scholar]
  53. 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]
  54. Treinin M., Laub O. Identification of a promoter element located upstream from the hepatitis B virus X gene. Mol Cell Biol. 1987 Jan;7(1):545–548. doi: 10.1128/mcb.7.1.545. [DOI] [PMC free article] [PubMed] [Google Scholar]
  55. Twu J. S., Chu K., Robinson W. S. Hepatitis B virus X gene activates kappa B-like enhancer sequences in the long terminal repeat of human immunodeficiency virus 1. Proc Natl Acad Sci U S A. 1989 Jul;86(13):5168–5172. doi: 10.1073/pnas.86.13.5168. [DOI] [PMC free article] [PubMed] [Google Scholar]
  56. 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]
  57. Twu J. S., Rosen C. A., Haseltine W. A., Robinson W. S. Identification of a region within the human immunodeficiency virus type 1 long terminal repeat that is essential for transactivation by the hepatitis B virus gene X. J Virol. 1989 Jun;63(6):2857–2860. doi: 10.1128/jvi.63.6.2857-2860.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  58. 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]
  59. Will H., Reiser W., Weimer T., Pfaff E., Büscher M., Sprengel R., Cattaneo R., Schaller H. Replication strategy of human hepatitis B virus. J Virol. 1987 Mar;61(3):904–911. doi: 10.1128/jvi.61.3.904-911.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  60. Wollersheim M., Debelka U., Hofschneider P. H. A transactivating function encoded in the hepatitis B virus X gene is conserved in the integrated state. Oncogene. 1988 Nov;3(5):545–552. [PubMed] [Google Scholar]
  61. Yaginuma K., Shirakata Y., Kobayashi M., Koike K. Hepatitis B virus (HBV) particles are produced in a cell culture system by transient expression of transfected HBV DNA. Proc Natl Acad Sci U S A. 1987 May;84(9):2678–2682. doi: 10.1073/pnas.84.9.2678. [DOI] [PMC free article] [PubMed] [Google Scholar]
  62. Yee J. K. A liver-specific enhancer in the core promoter region of human hepatitis B virus. Science. 1989 Nov 3;246(4930):658–661. doi: 10.1126/science.2554495. [DOI] [PubMed] [Google Scholar]
  63. Yokosuka O., Omata M., Imazeki F., Ito Y., Okuda K. Hepatitis B virus RNA transcripts and DNA in chronic liver disease. N Engl J Med. 1986 Nov 6;315(19):1187–1192. doi: 10.1056/NEJM198611063151903. [DOI] [PubMed] [Google Scholar]
  64. Zahm P., Hofschneider P. H., Koshy R. The HBV X-ORF encodes a transactivator: a potential factor in viral hepatocarcinogenesis. Oncogene. 1988 Aug;3(2):169–177. [PubMed] [Google Scholar]
  65. de Wet J. R., Wood K. V., DeLuca M., Helinski D. R., Subramani S. Firefly luciferase gene: structure and expression in mammalian cells. Mol Cell Biol. 1987 Feb;7(2):725–737. doi: 10.1128/mcb.7.2.725. [DOI] [PMC free article] [PubMed] [Google Scholar]
  66. van Zonneveld A. J., Curriden S. A., Loskutoff D. J. Type 1 plasminogen activator inhibitor gene: functional analysis and glucocorticoid regulation of its promoter. Proc Natl Acad Sci U S A. 1988 Aug;85(15):5525–5529. doi: 10.1073/pnas.85.15.5525. [DOI] [PMC free article] [PubMed] [Google Scholar]

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