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. 1980 Feb;33(2):795–806. doi: 10.1128/jvi.33.2.795-806.1980

State of hepatitis B viral DNA in a human hepatoma cell line.

P L Marion, F H Salazar, J J Alexander, W S Robinson
PMCID: PMC288606  PMID: 6251250

Abstract

PLC/PRF/5, a tissue culture cell line isolated from a human hepatocellular carcinoma and producing hepatitis B surface antigen, was studied for the presence of hepatitis B virus (HBV)-specific DNA and RNA. PLC/PRF/5 cell DNA accelerated the rate of reassociation of HBV [32P]DNA, and quantitative experiments indicated that the cells contained approximately four copies of viral DNA per haploid, mammalian cell DNA equivalent. PLC/PRF/5 DNA accelerated the rate of reassociation of all individual restriction endonucleases HincII and HaeIII fragments of HBV [32P]DNA, indicating that DNA from all regions of the viral genome is present in the cells. This suggests that these cells contain at least most, and possibly all, of the viral genome. Digestion of PLC/PRF/5 cell DNA with restriction endonuclease HindIII (an enzyme found not to cleave the DNA of any HBV isolate so far examined) yielded only three fragments, all larger than virion DNA, which contained HBV DNA base sequences, suggesting that HBV DNA is integrated in high-molecular-weight DNA at three different sites in these cells and that there is no viral DNA in an episomal form. PLC/PRF/5 cell [32P]RNA was found to hybridize with all restriction fragments of HBV DNA adequately tested, indicating that at least most, and possibly all, of the viral DNA in these cells is transcribed.

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Selected References

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

  1. Almeida J. D., Rubenstein D., Stott E. J. New antigen-antibody system in Australia-antigen-positive hepatitis. Lancet. 1971 Dec 4;2(7736):1225–1227. doi: 10.1016/s0140-6736(71)90543-5. [DOI] [PubMed] [Google Scholar]
  2. Alter H. J., Blumberg B. S. Further studies on a "new" human isoprecipitin system (Australia antigen). Blood. 1966 Mar;27(3):297–309. [PubMed] [Google Scholar]
  3. Dane D. S., Cameron C. H., Briggs M. Virus-like particles in serum of patients with Australia-antigen-associated hepatitis. Lancet. 1970 Apr 4;1(7649):695–698. doi: 10.1016/s0140-6736(70)90926-8. [DOI] [PubMed] [Google Scholar]
  4. Hirt B. Selective extraction of polyoma DNA from infected mouse cell cultures. J Mol Biol. 1967 Jun 14;26(2):365–369. doi: 10.1016/0022-2836(67)90307-5. [DOI] [PubMed] [Google Scholar]
  5. Hruska J. F., Clayton D. A., Rubenstein J. L., Robinson W. S. Structure of hepatitis B Dane particle DNA before and after the Dane particle DNA polymerase reaction. J Virol. 1977 Feb;21(2):666–672. doi: 10.1128/jvi.21.2.666-672.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Kaplan P. M., Greenman R. L., Gerin J. L., Purcell R. H., Robinson W. S. DNA polymerase associated with human hepatitis B antigen. J Virol. 1973 Nov;12(5):995–1005. doi: 10.1128/jvi.12.5.995-1005.1973. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Landers T. A., Greenberg H. B., Robinson W. S. Structure of hepatitis B Dane particle DNA and nature of the endogenous DNA polymerase reaction. J Virol. 1977 Aug;23(2):368–376. doi: 10.1128/jvi.23.2.368-376.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Lutwick L. I., Robinson W. S. DNA synthesized in the hepatitis B Dane particle DNA polymerase reaction. J Virol. 1977 Jan;21(1):96–104. doi: 10.1128/jvi.21.1.96-104.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. 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]
  10. 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]
  11. Rigby P. W., Berg P. Does simian virus 40 DNA integrate into cellular DNA during productive infection? J Virol. 1978 Nov;28(2):475–489. doi: 10.1128/jvi.28.2.475-489.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. 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]
  13. 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]
  14. Robinson W. S. The genome of hepatitis B virus. Annu Rev Microbiol. 1977;31:357–377. doi: 10.1146/annurev.mi.31.100177.002041. [DOI] [PubMed] [Google Scholar]
  15. Siddiqui A., Sattler F., Robinson W. S. Restriction endonuclease cleavage map and location of unique features of the DNA of hepatitis B virus, subtype adw2. Proc Natl Acad Sci U S A. 1979 Sep;76(9):4664–4668. doi: 10.1073/pnas.76.9.4664. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Sninsky J. J., Siddiqui A., Robinson W. S., Cohen S. N. Cloning and endonuclease mapping of the hepatitis B viral genome. Nature. 1979 May 24;279(5711):346–348. doi: 10.1038/279346a0. [DOI] [PubMed] [Google Scholar]
  17. Soulier J. P. Australia antigen subtypes. Discussion. Bibl Haematol. 1976;42:128–136. [PubMed] [Google Scholar]
  18. 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]
  19. Summers J., O'Connell A., Millman I. Genome of hepatitis B virus: restriction enzyme cleavage and structure of DNA extracted from Dane particles. Proc Natl Acad Sci U S A. 1975 Nov;72(11):4597–4601. doi: 10.1073/pnas.72.11.4597. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Summers J., Smolec J. M., Snyder R. A virus similar to human hepatitis B virus associated with hepatitis and hepatoma in woodchucks. Proc Natl Acad Sci U S A. 1978 Sep;75(9):4533–4537. doi: 10.1073/pnas.75.9.4533. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Szmuness W. Hepatocellular carcinoma and the hepatitis B virus: evidence for a causal association. Prog Med Virol. 1978;24:40–69. [PubMed] [Google Scholar]
  22. Takahashi K., Akahane Y., Gotanda T., Mishiro T., Imai M., Miyakawa Y., Mayumi M. Demonstration of hepatitis B e antigen in the core of Dane particles. J Immunol. 1979 Jan;122(1):275–279. [PubMed] [Google Scholar]

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