Skip to main content
NCBI home page
Journal of Virology logoLink to Journal of Virology
. 1992 Sep;66(9):5284–5289. doi: 10.1128/jvi.66.9.5284-5289.1992

The hepatitis B virus pre-S/S(t) transactivator is generated by 3' truncations within a defined region of the S gene.

U Lauer 1, L Weiss 1, P H Hofschneider 1, A S Kekulé 1
PMCID: PMC289082  PMID: 1323694

Abstract

Recently, it was reported that 3' truncation of an integrated surface gene (pre-S2/S) cloned from a hepatitis B virus (HBV)-associated hepatoma gave rise to the generation of a C-terminally truncated middle surface protein (MHBst), which surprisingly exerted a transcriptional transactivator function. To define the molecular requirements for the generation of transactivating MHBs(t) proteins, a 3' deletion analysis of the HBV pre-S2/S gene was performed. In cotransfection experiments with reporter plasmid pSV2CAT, full-length pre-S2/S genes or pre-S2/S genes with minor 3'-terminal deletions did not exhibit transactivator activity. In contrast, deletion of C-terminal hydrophobic region III of the S domain generated the transactivator function. Further stepwise 3' deletions, removing hydrophobic region II and both hydrophilic regions of the S domain, did not interfere with the transactivator activity; it was completely abolished, however, after additional deletion of hydrophobic region I. The results of this study define a range within the S open reading frame (between HBV nucleotides 221 and 573), termed the trans-activity-on region, in which 3' deletions give rise to the generation of transactivating MHBs(t) proteins. Within this region, not only 3' deletions but also the introduction of a stop codon activated the transactivator function, indicating that point mutations of integrated HBV DNA also may give rise to the synthesis of transactivating MHBs(t) proteins in vivo.

Full text

PDF
5284

Images in this article

5286Image
on p.5286

Selected References

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

  1. Angel P., Imagawa M., Chiu R., Stein B., Imbra R. J., Rahmsdorf H. J., Jonat C., Herrlich P., Karin M. Phorbol ester-inducible genes contain a common cis element recognized by a TPA-modulated trans-acting factor. Cell. 1987 Jun 19;49(6):729–739. doi: 10.1016/0092-8674(87)90611-8. [DOI] [PubMed] [Google Scholar]
  2. Berger I., Shaul Y. Integration of hepatitis B virus: analysis of unoccupied sites. J Virol. 1987 Apr;61(4):1180–1186. doi: 10.1128/jvi.61.4.1180-1186.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. 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]
  4. Chirgwin J. M., Przybyla A. E., MacDonald R. J., Rutter W. J. Isolation of biologically active ribonucleic acid from sources enriched in ribonuclease. Biochemistry. 1979 Nov 27;18(24):5294–5299. doi: 10.1021/bi00591a005. [DOI] [PubMed] [Google Scholar]
  5. Choo K. B., Liu M. S., Chang P. C., Wu S. M., Su M. W., Pan C. C., Han S. H. Analysis of six distinct integrated hepatitis B virus sequences cloned from the cellular DNA of a human hepatocellular carcinoma. Virology. 1986 Oct 30;154(2):405–408. doi: 10.1016/0042-6822(86)90467-8. [DOI] [PubMed] [Google Scholar]
  6. Eble B. E., Lingappa V. R., Ganem D. The N-terminal (pre-S2) domain of a hepatitis B virus surface glycoprotein is translocated across membranes by downstream signal sequences. J Virol. 1990 Mar;64(3):1414–1419. doi: 10.1128/jvi.64.3.1414-1419.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Eble B. E., MacRae D. R., Lingappa V. R., Ganem D. Multiple topogenic sequences determine the transmembrane orientation of the hepatitis B surface antigen. Mol Cell Biol. 1987 Oct;7(10):3591–3601. doi: 10.1128/mcb.7.10.3591. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Galibert F., Mandart E., Fitoussi F., Tiollais P., Charnay P. Nucleotide sequence of the hepatitis B virus genome (subtype ayw) cloned in E. coli. Nature. 1979 Oct 25;281(5733):646–650. doi: 10.1038/281646a0. [DOI] [PubMed] [Google Scholar]
  9. 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]
  10. 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]
  11. Graham F. L., van der Eb A. J. A new technique for the assay of infectivity of human adenovirus 5 DNA. Virology. 1973 Apr;52(2):456–467. doi: 10.1016/0042-6822(73)90341-3. [DOI] [PubMed] [Google Scholar]
  12. Grässer F. A., Graf T., Lipsick J. S. Protein truncation is required for the activation of the c-myb proto-oncogene. Mol Cell Biol. 1991 Aug;11(8):3987–3996. doi: 10.1128/mcb.11.8.3987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Imai M., Hoshi Y., Okamoto H., Matsui T., Tsurimoto T., Matsubara K., Miyakawa Y., Mayumi M. Free and integrated forms of hepatitis B virus DNA in human hepatocellular carcinoma cells (PLC/342) propagated in nude mice. J Virol. 1987 Nov;61(11):3555–3560. doi: 10.1128/jvi.61.11.3555-3560.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Kekulé A. S., Lauer U., Meyer M., Caselmann W. H., Hofschneider P. H., Koshy R. The preS2/S region of integrated hepatitis B virus DNA encodes a transcriptional transactivator. Nature. 1990 Feb 1;343(6257):457–461. doi: 10.1038/343457a0. [DOI] [PubMed] [Google Scholar]
  15. Koch S., von Loringhoven A. F., Hofschneider P. H., Koshy R. Amplification and rearrangement in hepatoma cell DNA associated with integrated hepatitis B virus DNA. EMBO J. 1984 Sep;3(9):2185–2189. doi: 10.1002/j.1460-2075.1984.tb02111.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Lüscher B., Eisenman R. N. New light on Myc and Myb. Part II. Myb. Genes Dev. 1990 Dec;4(12B):2235–2241. doi: 10.1101/gad.4.12b.2235. [DOI] [PubMed] [Google Scholar]
  17. Marschall M., Motz M., Leser U., Schwarzmann F., Oker B., Wolf H. Hepatitis B virus surface antigen as a reporter of promoter activity. Gene. 1989 Sep 1;81(1):109–117. doi: 10.1016/0378-1119(89)90341-7. [DOI] [PubMed] [Google Scholar]
  18. Nagaya T., Nakamura T., Tokino T., Tsurimoto T., Imai M., Mayumi T., Kamino K., Yamamura K., Matsubara K. The mode of hepatitis B virus DNA integration in chromosomes of human hepatocellular carcinoma. Genes Dev. 1987 Oct;1(8):773–782. doi: 10.1101/gad.1.8.773. [DOI] [PubMed] [Google Scholar]
  19. Peterson D. L. The structure of hepatitis B surface antigen and its antigenic sites. Bioessays. 1987 Jun;6(6):258–262. doi: 10.1002/bies.950060604. [DOI] [PubMed] [Google Scholar]
  20. 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]
  21. Shaul Y., Garcia P. D., Schonberg S., Rutter W. J. Integration of hepatitis B virus DNA in chromosome-specific satellite sequences. J Virol. 1986 Sep;59(3):731–734. doi: 10.1128/jvi.59.3.731-734.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Shih C., Burke K., Chou M. J., Zeldis J. B., Yang C. S., Lee C. S., Isselbacher K. J., Wands J. R., Goodman H. M. Tight clustering of human hepatitis B virus integration sites in hepatomas near a triple-stranded region. J Virol. 1987 Nov;61(11):3491–3498. doi: 10.1128/jvi.61.11.3491-3498.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. 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]
  24. Takahashi N., Kobayashi I. Evidence for the double-strand break repair model of bacteriophage lambda recombination. Proc Natl Acad Sci U S A. 1990 Apr;87(7):2790–2794. doi: 10.1073/pnas.87.7.2790. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. 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]
  26. 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]
  27. 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]
  28. Will H., Cattaneo R., Darai G., Deinhardt F., Schellekens H., Schaller H. Infectious hepatitis B virus from cloned DNA of known nucleotide sequence. Proc Natl Acad Sci U S A. 1985 Feb;82(3):891–895. doi: 10.1073/pnas.82.3.891. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. 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]
  30. 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]
  31. Zhou Y. Z., Slagle B. L., Donehower L. A., vanTuinen P., Ledbetter D. H., Butel J. S. Structural analysis of a hepatitis B virus genome integrated into chromosome 17p of a human hepatocellular carcinoma. J Virol. 1988 Nov;62(11):4224–4231. doi: 10.1128/jvi.62.11.4224-4231.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Ziemer M., Garcia P., Shaul Y., Rutter W. J. Sequence of hepatitis B virus DNA incorporated into the genome of a human hepatoma cell line. J Virol. 1985 Mar;53(3):885–892. doi: 10.1128/jvi.53.3.885-892.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]

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

RESOURCES