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. 1977 Aug;23(2):368–376. doi: 10.1128/jvi.23.2.368-376.1977

Structure of Hepatitis B Dane Particle DNA and Nature of the Endogenous DNA Polymerase Reaction

Terry A Landers a,1, Harry B Greenberg a,2, William S Robinson a
PMCID: PMC515839  PMID: 69727

Abstract

The circular DNA of hepatitis B Dane particles, which serves as the primer/template for an endogenous DNA polymerase, was analyzed by electrophoresis before and after a polymerase reaction and after digestion by restriction endonuclease or single-strand-specific endonuclease S1. The unreacted molecules extracted from the particles were electrophoretically heterogeneous, and treatment with S1 nuclease produced double-stranded linear DNA ranging in length from 1,700 to 2,800 base pairs (bp). After an endogenous DNA polymerase reaction, two discrete species of DNA molecules were found: a circular form and a linear form 3,200 bp long. The reaction resulted in a population of molecules with an elongated and more homogeneous double-stranded region. These results suggest that the circular molecules in Dane particles have single-stranded regions of varying lengths that are made double stranded during the DNA polymerase reaction. The endogenous DNA polymerase was found to initiate apparently at random in a region spanning more than a third of the molecule. Analysis of restriction endonuclease cleavage fragments of the fully elongated DNA revealed that although the molecules were of a uniform length, they were somewhat heterogeneous in sequence. The sum of the sizes of the 10 major endonuclease Hae III-generated fragments, detected by ethidium bromide, was 3,880 bp. Two additional fragments (B and G) detected by autoradiography after an endogenous DNA polymerase reaction with 32P-labeled deoxynucleoside triphosphates made the total 4,910 bp.

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Selected References

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  1. Almeida J. D. Individual morphological variations seen in Australia antigen positive sera. Am J Dis Child. 1972 Apr;123(4):303–309. doi: 10.1001/archpedi.1972.02110100035013. [DOI] [PubMed] [Google Scholar]
  2. 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]
  3. Alter H. J., Seeff L. B., Kaplan P. M., McAuliffe V. J., Wright E. C., Gerin J. L., Purcell R. H., Holland P. V., Zimmerman H. J. Type B hepatitis: the infectivity of blood positive for e antigen and DNA polymerase after accidental needlestick exposure. N Engl J Med. 1976 Oct 21;295(17):909–913. doi: 10.1056/NEJM197610212951701. [DOI] [PubMed] [Google Scholar]
  4. Barker L. F., Murray R. Relationship of virus dose to incubation time of clinical hepatitis and time of appearance of hepatitis--associated antigen. Am J Med Sci. 1972 Jan;263(1):27–33. doi: 10.1097/00000441-197201000-00005. [DOI] [PubMed] [Google Scholar]
  5. 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]
  6. 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]
  7. 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]
  8. Krugman S., Hoofnagle J. H., Gerety R. J., Kaplan P. M., Gerin J. L. Viral hepatitis, type B, DNA polymerase activity and antibody to hepatitis B core antigen. N Engl J Med. 1974 Jun 13;290(24):1331–1335. doi: 10.1056/NEJM197406132902401. [DOI] [PubMed] [Google Scholar]
  9. Krugman S. Viral hepatitis type B: prospects for active immunization. Am J Med Sci. 1975 Sep-Oct;270(2):391–393. doi: 10.1097/00000441-197509000-00023. [DOI] [PubMed] [Google Scholar]
  10. MCCOLLUM R. W. The size of serum hepatitis virus. Proc Soc Exp Biol Med. 1952 Oct;81(1):157–160. doi: 10.3181/00379727-81-19809. [DOI] [PubMed] [Google Scholar]
  11. Middleton J. H., Edgell M. H., Hutchison C. A., 3rd Specific fragments of phi X174 deoxyribonucleic acid produced by a restriction enzyme from Haemophilus aegyptius, endonuclease Z. J Virol. 1972 Jul;10(1):42–50. doi: 10.1128/jvi.10.1.42-50.1972. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Okada K., Kamiyama I., Inomata M., Imai M., Miyakawa Y. e antigen and anti-e in the serum of asymptomatic carrier mothers as indicators of positive and negative transmission of hepatitis B virus to their infants. N Engl J Med. 1976 Apr 1;294(14):746–749. doi: 10.1056/NEJM197604012941402. [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. DNA and DNA polymerase in the core of the Dane particle of hepatitis B. Am J Med Sci. 1975 Jul-Aug;270(1):151–159. doi: 10.1097/00000441-197507000-00021. [DOI] [PubMed] [Google Scholar]
  15. Robinson W. S., Greenman R. L. DNA polymerase in the core of the human hepatitis B virus candidate. J Virol. 1974 Jun;13(6):1231–1236. doi: 10.1128/jvi.13.6.1231-1236.1974. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Robinson W. S., Lutwick L. I. The virus of hepatitis, type B (first of two parts). N Engl J Med. 1976 Nov 18;295(21):1168–1175. doi: 10.1056/NEJM197611182952105. [DOI] [PubMed] [Google Scholar]
  17. Shenk T. E., Rhodes C., Rigby P. W., Berg P. Biochemical method for mapping mutational alterations in DNA with S1 nuclease: the location of deletions and temperature-sensitive mutations in simian virus 40. Proc Natl Acad Sci U S A. 1975 Mar;72(3):989–993. doi: 10.1073/pnas.72.3.989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Smith H. O., Wilcox K. W. A restriction enzyme from Hemophilus influenzae. I. Purification and general properties. J Mol Biol. 1970 Jul 28;51(2):379–391. doi: 10.1016/0022-2836(70)90149-x. [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]

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