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. 1985 Mar;53(3):885–892. doi: 10.1128/jvi.53.3.885-892.1985

Sequence of hepatitis B virus DNA incorporated into the genome of a human hepatoma cell line.

M Ziemer, P Garcia, Y Shaul, W J Rutter
PMCID: PMC254723  PMID: 2983098

Abstract

Seven copies of integrated hepatitis B virus (HBV) DNA and contiguous genomic DNA from a human hepatoma cell line (PLC/PRF/5) have been isolated by molecular cloning and have been partially sequenced. The HBV sequences are fragmented and rearranged. Thus, the surface antigen gene is the only intact HBV transcription unit present in these integrated sequences. The sites of integration-recombination are dispersed over the entire viral genome; there is some preference for integration within the double-stranded portion of the genome. There are no repeats at the ends of the integrated HBV DNA fragments. Thus, recombination does not take place in a manner resembling the integration of retroviruses. The sequence data suggest that each HBV fragment is of the adw subtype. However, the integrated DNAs show an unexpected degree of sequence divergence. Direct evidence for the duplication, transposition, and subsequent divergence of two sequences is presented. The data surprisingly suggest that infection-integration of four distinct adw strains occurred.

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Selected References

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

  1. Beasley R. P., Hwang L. Y., Lin C. C., Chien C. S. Hepatocellular carcinoma and hepatitis B virus. A prospective study of 22 707 men in Taiwan. Lancet. 1981 Nov 21;2(8256):1129–1133. doi: 10.1016/s0140-6736(81)90585-7. [DOI] [PubMed] [Google Scholar]
  2. Bell G. I., Selby M. J., Rutter W. J. The highly polymorphic region near the human insulin gene is composed of simple tandemly repeating sequences. Nature. 1982 Jan 7;295(5844):31–35. doi: 10.1038/295031a0. [DOI] [PubMed] [Google Scholar]
  3. 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]
  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. Couroucé-Pauty A. M., Plançon A., Soulier J. P. Distribution of HBsAg subtypes in the world. Vox Sang. 1983;44(4):197–211. doi: 10.1111/j.1423-0410.1983.tb01885.x. [DOI] [PubMed] [Google Scholar]
  6. Dejean A., Brechot C., Tiollais P., Wain-Hobson S. Characterization of integrated hepatitis B viral DNA cloned from a human hepatoma and the hepatoma-derived cell line PLC/PRF/5. Proc Natl Acad Sci U S A. 1983 May;80(9):2505–2509. doi: 10.1073/pnas.80.9.2505. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Deuring R., Winterhoff U., Tamanoi F., Stabel S., Doerfler W. Site of linkage between adenovirus type 12 and cell DNAs in hamster tumour line CLAC3. Nature. 1981 Sep 3;293(5827):81–84. doi: 10.1038/293081a0. [DOI] [PubMed] [Google Scholar]
  8. Edman J. C., Gray P., Valenzuela P., Rall L. B., Rutter W. J. Integration of hepatitis B virus sequences and their expression in a human hepatoma cell. Nature. 1980 Jul 31;286(5772):535–538. doi: 10.1038/286535a0. [DOI] [PubMed] [Google Scholar]
  9. Fujiyama A., Miyanohara A., Nozaki C., Yoneyama T., Ohtomo N., Matsubara K. Cloning and structural analyses of hepatitis B virus DNAs, subtype adr. Nucleic Acids Res. 1983 Jul 11;11(13):4601–4610. doi: 10.1093/nar/11.13.4601. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Galibert F., Chen T. N., Mandart E. Nucleotide sequence of a cloned woodchuck hepatitis virus genome: comparison with the hepatitis B virus sequence. J Virol. 1982 Jan;41(1):51–65. doi: 10.1128/jvi.41.1.51-65.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Gutai M. W., Nathans D. Evolutionary variants of simian virus 40: Cellular DNA sequences and sequences at recombinant joints of substituted variants. J Mol Biol. 1978 Dec 5;126(2):275–288. doi: 10.1016/0022-2836(78)90363-7. [DOI] [PubMed] [Google Scholar]
  12. Hayday A., Ruley H. E., Fried M. Structural and biological analysis of integrated polyoma virus DNA and its adjacent host sequences cloned from transformed rat cells. J Virol. 1982 Oct;44(1):67–77. doi: 10.1128/jvi.44.1.67-77.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Kam W., Rall L. B., Smuckler E. A., Schmid R., Rutter W. J. Hepatitis B viral DNA in liver and serum of asymptomatic carriers. Proc Natl Acad Sci U S A. 1982 Dec;79(23):7522–7526. doi: 10.1073/pnas.79.23.7522. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. 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]
  15. MacNab G. M., Alexander J. J., Lecatsas G., Bey E. M., Urbanowicz J. M. Hepatitis B surface antigen produced by a human hepatoma cell line. Br J Cancer. 1976 Nov;34(5):509–515. doi: 10.1038/bjc.1976.205. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Marion P. L., Salazar F. H., Alexander J. J., Robinson W. S. Polypeptides of hepatitis B virus surface antigen produced by a hepatoma cell line. J Virol. 1979 Dec;32(3):796–802. doi: 10.1128/jvi.32.3.796-802.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Ogston C. W., Jonak G. J., Rogler C. E., Astrin S. M., Summers J. Cloning and structural analysis of integrated woodchuck hepatitis virus sequences from hepatocellular carcinomas of woodchucks. Cell. 1982 Jun;29(2):385–394. doi: 10.1016/0092-8674(82)90155-6. [DOI] [PubMed] [Google Scholar]
  18. 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]
  19. Pasek M., Goto T., Gilbert W., Zink B., Schaller H., MacKay P., Leadbetter G., Murray K. Hepatitis B virus genes and their expression in E. coli. Nature. 1979 Dec 6;282(5739):575–579. doi: 10.1038/282575a0. [DOI] [PubMed] [Google Scholar]
  20. Rall L. B., Standring D. N., Laub O., Rutter W. J. Transcription of hepatitis B virus by RNA polymerase II. Mol Cell Biol. 1983 Oct;3(10):1766–1773. doi: 10.1128/mcb.3.10.1766. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Reanney D. Genetic noise in evolution? 1984 Jan 26-Feb 1Nature. 307(5949):318–319. doi: 10.1038/307318a0. [DOI] [PubMed] [Google Scholar]
  22. Ruley H. E., Fried M. Clustered illegitimate recombination events in mammalian cells involving very short sequence homologies. Nature. 1983 Jul 14;304(5922):181–184. doi: 10.1038/304181a0. [DOI] [PubMed] [Google Scholar]
  23. Ruley H. E., Lania L., Chaudry F., Fried M. Use of a cellular polyadenylation signal by viral transcripts in polyoma virus transformed cells. Nucleic Acids Res. 1982 Aug 11;10(15):4515–4524. doi: 10.1093/nar/10.15.4515. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. 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]
  25. Shafritz D. A., Kew M. C. Identification of integrated hepatitis B virus DNA sequences in human hepatocellular carcinomas. Hepatology. 1981 Jan-Feb;1(1):1–8. doi: 10.1002/hep.1840010102. [DOI] [PubMed] [Google Scholar]
  26. Shaul Y., Ziemer M., Garcia P. D., Crawford R., Hsu H., Valenzuela P., Rutter W. J. Cloning and analysis of integrated hepatitis virus sequences from a human hepatoma cell line. J Virol. 1984 Sep;51(3):776–787. doi: 10.1128/jvi.51.3.776-787.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Stabel S., Doerfler W. Nucleotide sequence at the site of junction between adenovirus type 12 DNA and repetitive hamster cell DNA in transformed cell line CLAC1. Nucleic Acids Res. 1982 Dec 20;10(24):8007–8023. doi: 10.1093/nar/10.24.8007. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Standring D. N., Rutter W. J., Varmus H. E., Ganem D. Transcription of the hepatitis B surface antigen gene in cultured murine cells initiates within the presurface region. J Virol. 1984 May;50(2):563–571. doi: 10.1128/jvi.50.2.563-571.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Stringer J. R. DNA sequence homology and chromosomal deletion at a site of SV40 DNA integration. Nature. 1982 Mar 25;296(5855):363–366. doi: 10.1038/296363a0. [DOI] [PubMed] [Google Scholar]
  30. Stringer J. R. Integrated simian virus 40 DNA: nucleotide sequences at cell-virus recombinant junctions. J Virol. 1981 May;38(2):671–679. doi: 10.1128/jvi.38.2.671-679.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Summers J., Mason W. S. Replication of the genome of a hepatitis B--like virus by reverse transcription of an RNA intermediate. Cell. 1982 Jun;29(2):403–415. doi: 10.1016/0092-8674(82)90157-x. [DOI] [PubMed] [Google Scholar]
  32. Szmuness W. Hepatocellular carcinoma and the hepatitis B virus: evidence for a causal association. Prog Med Virol. 1978;24:40–69. [PubMed] [Google Scholar]
  33. Ullrich A., Dull T. J., Gray A., Brosius J., Sures I. Genetic variation in the human insulin gene. Science. 1980 Aug 1;209(4456):612–615. doi: 10.1126/science.6248962. [DOI] [PubMed] [Google Scholar]

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