Skip to main content
Journal of Virology logoLink to Journal of Virology
. 1989 Sep;63(9):3919–3925. doi: 10.1128/jvi.63.9.3919-3925.1989

Characterization of hepatitis B virus major surface antigen gene transcriptional regulatory elements in differentiated hepatoma cell lines.

A K Raney 1, D R Milich 1, A McLachlan 1
PMCID: PMC250988  PMID: 2548006

Abstract

The regulatory DNA sequence elements that control the expression of the hepatitis B virus major surface antigen gene in the hepatoblastoma cell line HepG2 were analyzed by using transient transfection assays. In this system, the hepatitis B virus enhancer increases transcription from the surface antigen promoter approximately twofold. The promoter elements regulating the expression of this gene are within a 200-nucleotide sequence located immediately upstream of the transcription initiation sites. The promoter consists of an 85-nucleotide distal element which increases transcription from the surface antigen gene by two- to fourfold and a proximal element of approximately 115 nucleotides which is essential for transcriptional activity. The proximal and distal promoter elements were shown to bind factors present in HepG2 nuclear extracts, which is consistent with the regulatory role demonstrated for these sequences. The regulatory role of these promoter sequences in the hepatocellular carcinoma cell lines PLC/PRF/5 and Hep3B was also demonstrated, indicating similar transcriptional regulation of the surface antigen gene in each of these differentiated hepatoma cell lines.

Full text

PDF
3919

Images in this article

Selected References

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

  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. Briggs M. R., Kadonaga J. T., Bell S. P., Tjian R. Purification and biochemical characterization of the promoter-specific transcription factor, Sp1. Science. 1986 Oct 3;234(4772):47–52. doi: 10.1126/science.3529394. [DOI] [PubMed] [Google Scholar]
  3. Bulla G. A., Siddiqui A. The hepatitis B virus enhancer modulates transcription of the hepatitis B virus surface antigen gene from an internal location. J Virol. 1988 Apr;62(4):1437–1441. doi: 10.1128/jvi.62.4.1437-1441.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. 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]
  5. 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]
  6. 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]
  7. Chang H. K., Chou C. K., Chang C., Su T. S., Hu C., Yoshida M., Ting L. P. The enhancer sequence of human hepatitis B virus can enhance the activity of its surface gene promoter. Nucleic Acids Res. 1987 Mar 11;15(5):2261–2268. doi: 10.1093/nar/15.5.2261. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Chomczynski P., Sacchi N. Single-step method of RNA isolation by acid guanidinium thiocyanate-phenol-chloroform extraction. Anal Biochem. 1987 Apr;162(1):156–159. doi: 10.1006/abio.1987.9999. [DOI] [PubMed] [Google Scholar]
  9. 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]
  10. Dignam J. D., Lebovitz R. M., Roeder R. G. Accurate transcription initiation by RNA polymerase II in a soluble extract from isolated mammalian nuclei. Nucleic Acids Res. 1983 Mar 11;11(5):1475–1489. doi: 10.1093/nar/11.5.1475. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. 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]
  12. Elfassi E. Broad specificity of the hepatitis B enhancer function. Virology. 1987 Sep;160(1):259–262. doi: 10.1016/0042-6822(87)90069-9. [DOI] [PubMed] [Google Scholar]
  13. Faktor O., De-Medina T., Shaul Y. Regulation of hepatitis B virus S gene promoter in transfected cell lines. Virology. 1988 Feb;162(2):362–368. doi: 10.1016/0042-6822(88)90476-x. [DOI] [PubMed] [Google Scholar]
  14. 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]
  15. Imazeki F., Yaginuma K., Omata M., Okuda K., Kobayashi M., Koike K. RNA transcripts of hepatitis B virus in hepatocellular carcinoma. Hepatology. 1987 Jul-Aug;7(4):753–757. doi: 10.1002/hep.1840070423. [DOI] [PubMed] [Google Scholar]
  16. 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]
  17. 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]
  18. 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]
  19. Linney E., Donerly S. DNA fragments from F9 PyEC mutants increase expression of heterologous genes in transfected F9 cells. Cell. 1983 Dec;35(3 Pt 2):693–699. doi: 10.1016/0092-8674(83)90102-2. [DOI] [PubMed] [Google Scholar]
  20. 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]
  21. 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]
  22. Robinson W. S., Clayton D. A., Greenman R. L. DNA of a human hepatitis B virus candidate. J Virol. 1974 Aug;14(2):384–391. doi: 10.1128/jvi.14.2.384-391.1974. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. 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]
  24. 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]
  25. 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]
  26. Shaul Y., Ben-Levy R., De-Medina T. High affinity binding site for nuclear factor I next to the hepatitis B virus S gene promoter. EMBO J. 1986 Aug;5(8):1967–1971. doi: 10.1002/j.1460-2075.1986.tb04451.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Shaul Y., Rutter W. J., Laub O. A human hepatitis B viral enhancer element. EMBO J. 1985 Feb;4(2):427–430. doi: 10.1002/j.1460-2075.1985.tb03646.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. 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]
  29. 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]
  30. 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]
  31. Sureau C., Eichberg J. W., Hubbard G. B., Romet-Lemonne J. L., Essex M. A molecularly cloned hepatitis B virus produced in vitro is infectious in a chimpanzee. J Virol. 1988 Aug;62(8):3064–3067. doi: 10.1128/jvi.62.8.3064-3067.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. 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]
  33. 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]
  34. Tognoni A., Cattaneo R., Serfling E., Schaffner W. A novel expression selection approach allows precise mapping of the hepatitis B virus enhancer. Nucleic Acids Res. 1985 Oct 25;13(20):7457–7472. doi: 10.1093/nar/13.20.7457. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. 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]
  36. Tsurimoto T., Fujiyama A., Matsubara K. Stable expression and replication of hepatitis B virus genome in an integrated state in a human hepatoma cell line transfected with the cloned viral DNA. Proc Natl Acad Sci U S A. 1987 Jan;84(2):444–448. doi: 10.1073/pnas.84.2.444. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. 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]
  38. Vannice J. L., Levinson A. D. Properties of the human hepatitis B virus enhancer: position effects and cell-type nonspecificity. J Virol. 1988 Apr;62(4):1305–1313. doi: 10.1128/jvi.62.4.1305-1313.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  39. 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]
  40. 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]
  41. 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]

Articles from Journal of Virology are provided here courtesy of American Society for Microbiology (ASM)

RESOURCES