Abstract
DNA polymerase activity was detected in each of eight preparations of concentrated human hepatitis B antigen (HBAg) rich in Dane particles prepared by high-speed centrifugation of antigen-positive human plasma and in none of seven control preparations prepared in the same way from HBAg-negative plasma. The incorporation of 3H-thymidine-methyl-5′-triphosphate into DNA was dependent on four deoxyribonucleoside triphosphates and MgCl2. Treatment of the concentrated HBAg preparations with the nonionic detergent Nonidet P-40 (NP40) more than doubled the enzyme activity. Fractionation of the concentrated HBAg preparation in sucrose density gradients after treatment with NP40 revealed that the enzyme activity appeared within the density range of Dane core antigen but at a slightly higher density than the average for core antigen. The only particles observed by electron microscopy in this region of the gradient were typical 28-nm cores, suggesting that the DNA polymerase activity was associated with a subpopulation of cores. No DNA polymerase activity was found in purified 20-nm HBAg particles. The DNA product of the reaction remained associated with the 110S core and was not susceptible to DNase digestion when associated with the core. Inhibition of the reaction by actinomycin D and daunomycin suggested that the reaction was dependent on a DNA template associated with the core.
Full text
PDFSelected References
These references are in PubMed. This may not be the complete list of references from this article.
- Almeida J. D. Electron microscopic observations and speculations on Australia antigen. Postgrad Med J. 1971 Jul;47(549):484–487. doi: 10.1136/pgmj.47.549.484. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Bayer M. E., Blumberg B. S., Werner B. Particles associated with Australia antigen in the sera of patients with leukaemia, Down's Syndrome and hepatitis. Nature. 1968 Jun 15;218(5146):1057–1059. doi: 10.1038/2181057a0. [DOI] [PubMed] [Google Scholar]
- Blumberg B. S., Gerstley B. J., Hungerford D. A., London W. T., Sutnick A. I. A serum antigen (Australia antigen) in Down's syndrome, leukemia, and hepatitis. Ann Intern Med. 1967 May;66(5):924–931. doi: 10.7326/0003-4819-66-5-924. [DOI] [PubMed] [Google Scholar]
- CRAWFORD L. V., BLACK P. H. THE NUCLEIC ACID OF SIMIAN VIRUS 40. Virology. 1964 Nov;24:388–392. doi: 10.1016/0042-6822(64)90176-x. [DOI] [PubMed] [Google Scholar]
- 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]
- Gerin J. L., Purcell R. H., Hoggan M. D., Holland P. V., Chanock R. M. Biophysical properties of Australia antigen. J Virol. 1969 Nov;4(5):763–768. doi: 10.1128/jvi.4.5.763-768.1969. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Ginsberg A. L., Conrad M. E., Bancroft W. H., Ling C. M., Overby L. R. Prevention of endemic HAA-positive hepatitis with gamma globulin. Use of a simple radioimmune assay to detect HAA. N Engl J Med. 1972 Mar 16;286(11):562–566. doi: 10.1056/NEJM197203162861102. [DOI] [PubMed] [Google Scholar]
- Goldberg I. H., Friedman P. A. Antibiotics and nucleic acids. Annu Rev Biochem. 1971;40:775–810. doi: 10.1146/annurev.bi.40.070171.004015. [DOI] [PubMed] [Google Scholar]
- Hirschman S. Z., Vernace S. J., Schaffner F. D.N.A. polymerase in preparations containing Australia antigen. Lancet. 1971 May 29;1(7709):1099–1103. doi: 10.1016/s0140-6736(71)91839-3. [DOI] [PubMed] [Google Scholar]
- Hung P. P., Robinson H. L., Robinson W. S. Isolation and characterization of proteins from Rous sarcoma virus. Virology. 1971 Jan;43(1):251–266. doi: 10.1016/0042-6822(71)90243-1. [DOI] [PubMed] [Google Scholar]
- Kates J. R., McAuslan B. R. Poxvirus DNA-dependent RNA polymerase. Proc Natl Acad Sci U S A. 1967 Jul;58(1):134–141. doi: 10.1073/pnas.58.1.134. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Krugman S., Giles J. P. Viral hepatitis. New light on an old disease. JAMA. 1970 May 11;212(6):1019–1029. doi: 10.1001/jama.212.6.1019. [DOI] [PubMed] [Google Scholar]
- LOWRY O. H., ROSEBROUGH N. J., FARR A. L., RANDALL R. J. Protein measurement with the Folin phenol reagent. J Biol Chem. 1951 Nov;193(1):265–275. [PubMed] [Google Scholar]
- Lander J. J., Alter H. J., Purcell R. H. Frequency of antibody to hepatitis-associated antigen as measured by a new radioimmunoassay technique. J Immunol. 1971 May;106(5):1166–1171. [PubMed] [Google Scholar]
- Lander J. J., Giles J. P., Purcell R. H., Krugman S. Viral hepatitis, type B (MS-2 strain). Detection of antibody after primary infection. N Engl J Med. 1971 Aug 5;285(6):303–307. doi: 10.1056/NEJM197108052850601. [DOI] [PubMed] [Google Scholar]
- Le Bouvier G. L. The heterogeneity of Australia antigen. J Infect Dis. 1971 Jun;123(6):671–675. doi: 10.1093/infdis/123.6.671. [DOI] [PubMed] [Google Scholar]
- Mosley J. W., Edwards V. M., Meihaus J. E., Redeker A. G. Subdeterminants d and y of hepatitis B antigen as epidemiologic markers. Am J Epidemiol. 1972 Jun;95(6):529–535. [PubMed] [Google Scholar]
- Okochi K., Murakami S. Observations on Australia antigen in Japanese. Vox Sang. 1968;15(5):374–385. doi: 10.1111/j.1423-0410.1968.tb04078.x. [DOI] [PubMed] [Google Scholar]
- Prince A. M. An antigen detected in the blood during the incubation period of serum hepatitis. Proc Natl Acad Sci U S A. 1968 Jul;60(3):814–821. doi: 10.1073/pnas.60.3.814. [DOI] [PMC free article] [PubMed] [Google Scholar]
- RICHARDSON C. C., INMAN R. B., KORNBERG A. ENZYMIC SYNTHESIS OF DEOXYRIBONUCLEIC ACID. 18. THE REPAIR OF PARTIALLY SINGLE-STRANDED DNA TEMPLATES BY DNA POLYMERASE. J Mol Biol. 1964 Jul;9:46–69. doi: 10.1016/s0022-2836(64)80090-5. [DOI] [PubMed] [Google Scholar]
- Reich E., Goldberg I. H. Actinomycin and nucleic acid function. Prog Nucleic Acid Res Mol Biol. 1964;3:183–234. doi: 10.1016/s0079-6603(08)60742-4. [DOI] [PubMed] [Google Scholar]
- Robinson W. S., Robinson H. L. DNA polymerase in defective Rous sarcoma virus. Virology. 1971 May;44(2):457–462. doi: 10.1016/0042-6822(71)90278-9. [DOI] [PubMed] [Google Scholar]
- Rosenthal P. N., Robinson H. L., Robinson W. S., Hanafusa T., Hanafusa H. DNA in uninfected and virus-infected cells complementary to avian tumor virus RNA. Proc Natl Acad Sci U S A. 1971 Oct;68(10):2336–2340. doi: 10.1073/pnas.68.10.2336. [DOI] [PMC free article] [PubMed] [Google Scholar]
- SCHACHMAN H. K., ADLER J., RADDING C. M., LEHMAN I. R., KORNBERG A. Enzymatic synthesis of deoxyribonucleic acid. VII. Synthesis of a polymer of deoxyadenylate and deoxythymidylate. J Biol Chem. 1960 Nov;235:3242–3249. [PubMed] [Google Scholar]
- Shulman N. R. Hepatitis-associated antigen. Am J Med. 1970 Nov;49:669–692. doi: 10.1016/S0002-9343(70)80133-4. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Temin H. M., Baltimore D. RNA-directed DNA synthesis and RNA tumor viruses. Adv Virus Res. 1972;17:129–186. doi: 10.1016/s0065-3527(08)60749-6. [DOI] [PubMed] [Google Scholar]
- Weissbach A., Bolden A., Muller R., Hanafusa H., Hanafusa T. Deoxyribonucleic acid polymerase activities in normal and leukovirus-infected chicken embryo cells. J Virol. 1972 Sep;10(3):321–327. doi: 10.1128/jvi.10.3.321-327.1972. [DOI] [PMC free article] [PubMed] [Google Scholar]