Skip to main content
Journal of Virology logoLink to Journal of Virology
. 1990 Feb;64(2):822–828. doi: 10.1128/jvi.64.2.822-828.1990

Structural rearrangement of integrated hepatitis B virus DNA as well as cellular flanking DNA is present in chronically infected hepatic tissues.

S Takada 1, Y Gotoh 1, S Hayashi 1, M Yoshida 1, K Koike 1
PMCID: PMC249177  PMID: 2296084

Abstract

Cellular DNAs from human livers chronically infected with hepatitis B virus (HBV) were analyzed by Southern blot hybridization for the presence of integrated HBV DNA. In 15 of 16 chronically infected hepatic tissues, random HBV DNA integration was evident. By molecular cloning and structural analyses of 19 integrants from three chronically infected hepatic tissues, deletion of cellular flanking DNA in all cases and rearrangement of HBV DNA with inverted duplication or translocation of cellular flanking DNA at the virus-cell junction in some cases were noted. Thus, the rearrangement of HBV DNA or cellular flanking DNA is not a specific incident of hepatocellular carcinoma formation. Detailed analyses of various integrants bearing rearranged viral DNA failed to indicate any gross structural alteration in cellular DNA, except for a small deletion at the integration site, indicating that viral DNA rearrangement with inverted duplication possibly occurs before integration of HBV DNA. Based on nucleotide sequencing analyses of virus-virus junctions, a one- to three-nucleotide identity was found. A mechanism for this inverted duplication of HBV DNA is proposed in which illegitimate recombination between two complementary viral strands may take place by means of a nucleotide identity at the junction site in a weakly homologous region (patchy homology) on one side of adjoining viral sequences. For virus-cell junctions, the mechanism may be basically similar to that for virus-virus junctions.

Full text

PDF
822

Images in this article

Selected References

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

  1. Benton W. D., Davis R. W. Screening lambdagt recombinant clones by hybridization to single plaques in situ. Science. 1977 Apr 8;196(4286):180–182. doi: 10.1126/science.322279. [DOI] [PubMed] [Google Scholar]
  2. Brechot C., Hadchouel M., Scotto J., Degos F., Charnay P., Trepo C., Tiollais P. Detection of hepatitis B virus DNA in liver and serum: a direct appraisal of the chronic carrier state. Lancet. 1981 Oct 10;2(8250):765–768. doi: 10.1016/s0140-6736(81)90182-3. [DOI] [PubMed] [Google Scholar]
  3. Bréchot C., Hadchouel M., Scotto J., Fonck M., Potet F., Vyas G. N., Tiollais P. State of hepatitis B virus DNA in hepatocytes of patients with hepatitis B surface antigen-positive and -negative liver diseases. Proc Natl Acad Sci U S A. 1981 Jun;78(6):3906–3910. doi: 10.1073/pnas.78.6.3906. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Chisari F. V., Filippi P., Buras J., McLachlan A., Popper H., Pinkert C. A., Palmiter R. D., Brinster R. L. Structural and pathological effects of synthesis of hepatitis B virus large envelope polypeptide in transgenic mice. Proc Natl Acad Sci U S A. 1987 Oct;84(19):6909–6913. doi: 10.1073/pnas.84.19.6909. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Dejean A., Sonigo P., Wain-Hobson S., Tiollais P. Specific hepatitis B virus integration in hepatocellular carcinoma DNA through a viral 11-base-pair direct repeat. Proc Natl Acad Sci U S A. 1984 Sep;81(17):5350–5354. doi: 10.1073/pnas.81.17.5350. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Hsu T., Möröy T., Etiemble J., Louise A., Trépo C., Tiollais P., Buendia M. A. Activation of c-myc by woodchuck hepatitis virus insertion in hepatocellular carcinoma. Cell. 1988 Nov 18;55(4):627–635. doi: 10.1016/0092-8674(88)90221-8. [DOI] [PubMed] [Google Scholar]
  7. Imazeki F., Yaginuma K., Omata M., Okuda K., Kobayashi M., Koike K. Integrated structures of duck hepatitis B virus DNA in hepatocellular carcinoma. J Virol. 1988 Mar;62(3):861–865. doi: 10.1128/jvi.62.3.861-865.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Kobayashi M., Koike K. Complete nucleotide sequence of hepatitis B virus DNA of subtype adr and its conserved gene organization. Gene. 1984 Oct;30(1-3):227–232. doi: 10.1016/0378-1119(84)90124-0. [DOI] [PubMed] [Google Scholar]
  9. Koch S., Freytag von Loringhoven A., Kahmann R., Hofschneider P. H., Koshy R. The genetic organization of integrated hepatitis B virus DNA in the human hepatoma cell line PLC/PRF/5. Nucleic Acids Res. 1984 Sep 11;12(17):6871–6886. doi: 10.1093/nar/12.17.6871. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Koike K., Kobayashi M., Mizusawa H., Yoshida E., Yaginuma K., Taira M. Rearrangement of the surface antigen gene of hepatitis B virus integrated in the human hepatoma cell lines. Nucleic Acids Res. 1983 Aug 25;11(16):5391–5402. doi: 10.1093/nar/11.16.5391. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Koshy R., Koch S., von Loringhoven A. F., Kahmann R., Murray K., Hofschneider P. H. Integration of hepatitis B virus DNA: evidence for integration in the single-stranded gap. Cell. 1983 Aug;34(1):215–223. doi: 10.1016/0092-8674(83)90152-6. [DOI] [PubMed] [Google Scholar]
  12. Maxam A. M., Gilbert W. Sequencing end-labeled DNA with base-specific chemical cleavages. Methods Enzymol. 1980;65(1):499–560. doi: 10.1016/s0076-6879(80)65059-9. [DOI] [PubMed] [Google Scholar]
  13. Mizusawa H., Taira M., Yaginuma K., Kobayashi M., Yoshida E., Koike K. Inversely repeating integrated hepatitis B virus DNA and cellular flanking sequences in the human hepatoma-derived cell line huSP. Proc Natl Acad Sci U S A. 1985 Jan;82(1):208–212. doi: 10.1073/pnas.82.1.208. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. 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]
  15. Ono Y., Onda H., Sasada R., Igarashi K., Sugino Y., Nishioka K. The complete nucleotide sequences of the cloned hepatitis B virus DNA; subtype adr and adw. Nucleic Acids Res. 1983 Mar 25;11(6):1747–1757. doi: 10.1093/nar/11.6.1747. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Popper H., Roth L., Purcell R. H., Tennant B. C., Gerin J. L. Hepatocarcinogenicity of the woodchuck hepatitis virus. Proc Natl Acad Sci U S A. 1987 Feb;84(3):866–870. doi: 10.1073/pnas.84.3.866. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Rigby P. W., Dieckmann M., Rhodes C., Berg P. Labeling deoxyribonucleic acid to high specific activity in vitro by nick translation with DNA polymerase I. J Mol Biol. 1977 Jun 15;113(1):237–251. doi: 10.1016/0022-2836(77)90052-3. [DOI] [PubMed] [Google Scholar]
  18. Rogler C. E., Sherman M., Su C. Y., Shafritz D. A., Summers J., Shows T. B., Henderson A., Kew M. Deletion in chromosome 11p associated with a hepatitis B integration site in hepatocellular carcinoma. Science. 1985 Oct 18;230(4723):319–322. doi: 10.1126/science.2996131. [DOI] [PubMed] [Google Scholar]
  19. Rogler C. E., Summers J. Cloning and structural analysis of integrated woodchuck hepatitis virus sequences from a chronically infected liver. J Virol. 1984 Jun;50(3):832–837. doi: 10.1128/jvi.50.3.832-837.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Rogler C. E., Summers J. Novel forms of woodchuck hepatitis virus DNA isolated from chronically infected woodchuck liver nuclei. J Virol. 1982 Dec;44(3):852–863. doi: 10.1128/jvi.44.3.852-863.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. 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]
  22. 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]
  23. Shafritz D. A., Shouval D., Sherman H. I., Hadziyannis S. J., Kew M. C. Integration of hepatitis B virus DNA into the genome of liver cells in chronic liver disease and hepatocellular carcinoma. Studies in percutaneous liver biopsies and post-mortem tissue specimens. N Engl J Med. 1981 Oct 29;305(18):1067–1073. doi: 10.1056/NEJM198110293051807. [DOI] [PubMed] [Google Scholar]
  24. 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]
  25. 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]
  26. 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]
  27. Szmuness W. Hepatocellular carcinoma and the hepatitis B virus: evidence for a causal association. Prog Med Virol. 1978;24:40–69. [PubMed] [Google Scholar]
  28. 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]
  29. Tokino T., Fukushige S., Nakamura T., Nagaya T., Murotsu T., Shiga K., Aoki N., Matsubara K. Chromosomal translocation and inverted duplication associated with integrated hepatitis B virus in hepatocellular carcinomas. J Virol. 1987 Dec;61(12):3848–3854. doi: 10.1128/jvi.61.12.3848-3854.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Yaginuma K., Kobayashi H., Kobayashi M., Morishima T., Matsuyama K., Koike K. Multiple integration site of hepatitis B virus DNA in hepatocellular carcinoma and chronic active hepatitis tissues from children. J Virol. 1987 Jun;61(6):1808–1813. doi: 10.1128/jvi.61.6.1808-1813.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Yaginuma K., Kobayashi M., Yoshida E., Koike K. Hepatitis B virus integration in hepatocellular carcinoma DNA: duplication of cellular flanking sequences at the integration site. Proc Natl Acad Sci U S A. 1985 Jul;82(13):4458–4462. doi: 10.1073/pnas.82.13.4458. [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