Skip to main content
NCBI home page
Journal of Virology logoLink to Journal of Virology
. 1991 Aug;65(8):4284–4291. doi: 10.1128/jvi.65.8.4284-4291.1991

Diversity of hepatitis B virus X gene-related transcripts in hepatocellular carcinoma: a novel polyadenylation site on viral DNA.

C Hilger 1, I Velhagen 1, H Zentgraf 1, C H Schröder 1
PMCID: PMC248866  PMID: 1649331

Abstract

RNA was isolated from tissue of two patients with hepatocellular carcinoma developed on the background of a chronic hepatitis B virus infection. For identification and characterization of 3' ends of X gene open reading frame (ORF)-related transcripts, RNA was reverse transcribed into cDNA and subjected to polymerase chain reaction. Cloned amplification products from tumor tissue of one patient represented an approximately even distribution of transcripts terminating at the established poly(A) signal (standard transcripts) and of truncated transcripts terminating at a CATAAA poly(A) signal within the 3' end region of X gene ORF (truncated transcripts). Amplified cDNA from tumor tissue of the second patient could be attributed mainly to the standard type of transcripts, whereas cDNA from the nontumor tissue of the same patient could be assigned to four groups of transcripts: (i) standard transcripts, (ii) transcripts with internal deletions affecting the 3' end of the X gene, (iii) truncated transcripts, and (iv) hybrid transcripts displaying 5' sequences from the X gene ORF fused to cellular sequences.

Full text

PDF
4284

Images in this article

4286Image
on p.4286

Selected References

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

  1. Acs G., Sells M. A., Purcell R. H., Price P., Engle R., Shapiro M., Popper H. Hepatitis B virus produced by transfected Hep G2 cells causes hepatitis in chimpanzees. Proc Natl Acad Sci U S A. 1987 Jul;84(13):4641–4644. doi: 10.1073/pnas.84.13.4641. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Artelt P., Morelle C., Ausmeier M., Fitzek M., Hauser H. Vectors for efficient expression in mammalian fibroblastoid, myeloid and lymphoid cells via transfection or infection. Gene. 1988 Sep 7;68(2):213–219. doi: 10.1016/0378-1119(88)90023-6. [DOI] [PubMed] [Google Scholar]
  3. Beasley R. P. Hepatitis B virus. The major etiology of hepatocellular carcinoma. Cancer. 1988 May 15;61(10):1942–1956. doi: 10.1002/1097-0142(19880515)61:10<1942::aid-cncr2820611003>3.0.co;2-j. [DOI] [PubMed] [Google Scholar]
  4. Bradford M. M. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem. 1976 May 7;72:248–254. doi: 10.1016/0003-2697(76)90527-3. [DOI] [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. Chen C., Okayama H. High-efficiency transformation of mammalian cells by plasmid DNA. Mol Cell Biol. 1987 Aug;7(8):2745–2752. doi: 10.1128/mcb.7.8.2745. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Faktor O., Shaul Y. The identification of hepatitis B virus X gene responsive elements reveals functional similarity of X and HTLV-I tax. Oncogene. 1990 Jun;5(6):867–872. [PubMed] [Google Scholar]
  8. Frohman M. A., Dush M. K., Martin G. R. Rapid production of full-length cDNAs from rare transcripts: amplification using a single gene-specific oligonucleotide primer. Proc Natl Acad Sci U S A. 1988 Dec;85(23):8998–9002. doi: 10.1073/pnas.85.23.8998. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. 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]
  10. Hino O., Ohtake K., Rogler C. E. Features of two hepatitis B virus (HBV) DNA integrations suggest mechanisms of HBV integration. J Virol. 1989 Jun;63(6):2638–2643. doi: 10.1128/jvi.63.6.2638-2643.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Hu K. Q., Vierling J. M., Siddiqui A. Trans-activation of HLA-DR gene by hepatitis B virus X gene product. Proc Natl Acad Sci U S A. 1990 Sep;87(18):7140–7144. doi: 10.1073/pnas.87.18.7140. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Höhne M., Schaefer S., Seifer M., Feitelson M. A., Paul D., Gerlich W. H. Malignant transformation of immortalized transgenic hepatocytes after transfection with hepatitis B virus DNA. EMBO J. 1990 Apr;9(4):1137–1145. doi: 10.1002/j.1460-2075.1990.tb08220.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. 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]
  14. Knowles B. B., Howe C. C., Aden D. P. Human hepatocellular carcinoma cell lines secrete the major plasma proteins and hepatitis B surface antigen. Science. 1980 Jul 25;209(4455):497–499. doi: 10.1126/science.6248960. [DOI] [PubMed] [Google Scholar]
  15. Konopka A. K. Compilation of DNA strand exchange sites for non-homologous recombination in somatic cells. Nucleic Acids Res. 1988 Mar 11;16(5):1739–1758. doi: 10.1093/nar/16.5.1739. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. 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]
  17. Lo S. J., You L. R., Lee Y. H. The hepatitis B virus X-C fusion protein is unlikely to be produced by the mechanism of ribosomal frameshifting. Virology. 1990 Oct;178(2):584–587. doi: 10.1016/0042-6822(90)90358-x. [DOI] [PubMed] [Google Scholar]
  18. Loncarevic I. F., Zentgraf H., Schröder C. H. Sequence of a replication competent hepatitis B virus genome with a preX open reading frame. Nucleic Acids Res. 1990 Aug 25;18(16):4940–4940. doi: 10.1093/nar/18.16.4940. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Loncarević I. F., Schranz P., Zentgraf H., Liang X. H., Herrmann G., Tang Z. Y., Schröder C. H. Replication of hepatitis B virus in a hepatocellular carcinoma. Virology. 1990 Jan;174(1):158–168. doi: 10.1016/0042-6822(90)90064-x. [DOI] [PubMed] [Google Scholar]
  20. Luckow B., Schütz G. CAT constructions with multiple unique restriction sites for the functional analysis of eukaryotic promoters and regulatory elements. Nucleic Acids Res. 1987 Jul 10;15(13):5490–5490. doi: 10.1093/nar/15.13.5490. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Mount S. M. A catalogue of splice junction sequences. Nucleic Acids Res. 1982 Jan 22;10(2):459–472. doi: 10.1093/nar/10.2.459. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. 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]
  23. Ou J., Rutter W. J. Hybrid hepatitis B virus-host transcripts in a human hepatoma cell. Proc Natl Acad Sci U S A. 1985 Jan;82(1):83–87. doi: 10.1073/pnas.82.1.83. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. 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]
  25. Radziwill G., Tucker W., Schaller H. Mutational analysis of the hepatitis B virus P gene product: domain structure and RNase H activity. J Virol. 1990 Feb;64(2):613–620. doi: 10.1128/jvi.64.2.613-620.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Saiki R. K., Gelfand D. H., Stoffel S., Scharf S. J., Higuchi R., Horn G. T., Mullis K. B., Erlich H. A. Primer-directed enzymatic amplification of DNA with a thermostable DNA polymerase. Science. 1988 Jan 29;239(4839):487–491. doi: 10.1126/science.2448875. [DOI] [PubMed] [Google Scholar]
  27. 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]
  28. Sheets M. D., Ogg S. C., Wickens M. P. Point mutations in AAUAAA and the poly (A) addition site: effects on the accuracy and efficiency of cleavage and polyadenylation in vitro. Nucleic Acids Res. 1990 Oct 11;18(19):5799–5805. doi: 10.1093/nar/18.19.5799. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Shirakata Y., Kawada M., Fujiki Y., Sano H., Oda M., Yaginuma K., Kobayashi M., Koike K. The X gene of hepatitis B virus induced growth stimulation and tumorigenic transformation of mouse NIH3T3 cells. Jpn J Cancer Res. 1989 Jul;80(7):617–621. doi: 10.1111/j.1349-7006.1989.tb01686.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. 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]
  31. Simonsen C. C., Levinson A. D. Analysis of processing and polyadenylation signals of the hepatitis B virus surface antigen gene by using simian virus 40-hepatitis B virus chimeric plasmids. Mol Cell Biol. 1983 Dec;3(12):2250–2258. doi: 10.1128/mcb.3.12.2250. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Southern E. M. Detection of specific sequences among DNA fragments separated by gel electrophoresis. J Mol Biol. 1975 Nov 5;98(3):503–517. doi: 10.1016/s0022-2836(75)80083-0. [DOI] [PubMed] [Google Scholar]
  33. 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]
  34. Takada S., Gotoh Y., Hayashi S., Yoshida M., Koike K. Structural rearrangement of integrated hepatitis B virus DNA as well as cellular flanking DNA is present in chronically infected hepatic tissues. J Virol. 1990 Feb;64(2):822–828. doi: 10.1128/jvi.64.2.822-828.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. 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]
  36. 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]
  37. 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]
  38. 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]
  39. 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]
  40. 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]
  41. Zhou D. X., Taraboulos A., Ou J. H., Yen T. S. Activation of class I major histocompatibility complex gene expression by hepatitis B virus. J Virol. 1990 Aug;64(8):4025–4028. doi: 10.1128/jvi.64.8.4025-4028.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  42. 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]
  43. von Loringhoven A. F., Koch S., Hofschneider P. H., Koshy R. Co-transcribed 3' host sequences augment expression of integrated hepatitis B virus DNA. EMBO J. 1985 Jan;4(1):249–255. doi: 10.1002/j.1460-2075.1985.tb02343.x. [DOI] [PMC free article] [PubMed] [Google Scholar]

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

RESOURCES