Skip to main content
NCBI home page
Journal of Virology logoLink to Journal of Virology
. 1978 Jan;25(1):459–463. doi: 10.1128/jvi.25.1.459-463.1978

Adenovirus DNA synthesis in vitro in an isolated complex.

G D Frenkel
PMCID: PMC353950  PMID: 202756

Abstract

DNA-protein complexes isolated from adenovirus-infected cells by a modification of the M-band technique were used as an in vitro system for the study of adenovirus DNA replication. The synthesis in vitro was semiconservative, inhibited by N-ethylmaleimide, and stimulated by ATP. Studies on DNA-negative mutants of adenovirus showed that the DNA synthesis in vitro represents a continuation of adenovirus DNA replication in vivo. DNA synthesis in vitro was inhibited 38% by 20 microgram of phosphonoacetic acid per ml, which is several-fold higher than the inhibition obtained with purified DNA polymerase beta or gamma, but was similar to the degree of inhibition of DNA polymerase alpha. DNA synthesis in complexes from uninfected cells was much less sensitive to inhibition by phosphonoacetic acid. In addition, complexes from infected cells contained a greater proportion of the alpha-polymerase than complexes from uninfected cells, suggesting that an association of alpha-polymerase with the replication complex may be occurring during adenovirus infection, with subsequent utilization of the alpha-polymerase for viral DNA synthesis.

Full text

PDF
459

Selected References

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

  1. Bolden A., Aucker J., Weissbach A. Synthesis of herpes simplex virus, vaccinia virus, and adenovirus DNA in isolated HeLa cell nuclei. I. Effect of viral-specific antisera and phosphonoacetic acid. J Virol. 1975 Dec;16(6):1584–1592. doi: 10.1128/jvi.16.6.1584-1592.1975. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Ensinger M. J., Ginsberg H. S. Selection and preliminary characterization of temperature-sensitive mutants of type 5 adenovirus. J Virol. 1972 Sep;10(3):328–339. doi: 10.1128/jvi.10.3.328-339.1972. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Ito K., Arens M., Green M. Characterization of DNA polymerase associated with nuclear membrane fractions from uninfected and adenovirus 2-infected KB cells. Biochim Biophys Acta. 1976 Oct 18;447(3):340–352. doi: 10.1016/0005-2787(76)90057-5. [DOI] [PubMed] [Google Scholar]
  4. Ito K., Arens M., Green M. Isolation of DNA polymerase gamma from an adenovirus 2 DNA replication complex. J Virol. 1975 Jun;15(6):1507–1510. doi: 10.1128/jvi.15.6.1507-1510.1975. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Levine A. J., van der Vliet P. C., Sussenbach J. S. The replication of papovavirus and adenovirus DNA. Curr Top Microbiol Immunol. 1976;73:67–124. doi: 10.1007/978-3-642-66306-2_3. [DOI] [PubMed] [Google Scholar]
  6. Lewis B. J., Abrell J. W., Smith R. G., Gallo R. C. DNA polymerases in human lymphoblastoid cells infected with simian sarcoma virus. Biochim Biophys Acta. 1974 May 17;349(2):148–160. doi: 10.1016/0005-2787(74)90076-8. [DOI] [PubMed] [Google Scholar]
  7. Moses R. E., Richardson C. C. Replication and repair of DNA in cells of Escherichia coli treated with toluene. Proc Natl Acad Sci U S A. 1970 Oct;67(2):674–681. doi: 10.1073/pnas.67.2.674. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Shiroki K., Shimojo H., Yamaguchi K. The viral DNA replication complex of adenovirus 12. Virology. 1974 Jul;60(1):192–199. doi: 10.1016/0042-6822(74)90376-6. [DOI] [PubMed] [Google Scholar]
  9. Spadari S., Weissbach A. HeLa cell R-deoxyribonucleic acid polymerases. Separation and characterization of two enzymatic activities. J Biol Chem. 1974 Sep 25;249(18):5809–5815. [PubMed] [Google Scholar]
  10. Spadari S., Weissbach A. The interrelation between DNA synthesis and various DNA polymerase activities in synchronized HeLa cells. J Mol Biol. 1974 Jun 15;86(1):11–20. doi: 10.1016/s0022-2836(74)80003-3. [DOI] [PubMed] [Google Scholar]
  11. Tremblay G. Y., Daniels M. J., Schaechter M. Isolation of a cell membrane-DNA-nascent RNA complex from bacteria. J Mol Biol. 1969 Feb 28;40(1):65–76. doi: 10.1016/0022-2836(69)90296-4. [DOI] [PubMed] [Google Scholar]
  12. Van Der Vliet P. C., Levine A. J., Ensinger M. J., Ginsberg H. S. Thermolabile DNA binding proteins from cells infected with a temperature-sensitive mutant of adenovrius defective in viral DNA synthesis. J Virol. 1975 Feb;15(2):348–354. doi: 10.1128/jvi.15.2.348-354.1975. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Vliet P. C., Sussenbach J. S. An adenovirus type 5 gene function required for initiation of viral DNA replication. Virology. 1975 Oct;67(2):415–426. doi: 10.1016/0042-6822(75)90443-2. [DOI] [PubMed] [Google Scholar]
  14. Wilkie N. M., Ustacelebi S., Williams J. F. Characterization of temperature-sensitive mutants of adenovirus type 5: nucleic acid synthesis. Virology. 1973 Feb;51(2):499–503. doi: 10.1016/0042-6822(73)90450-9. [DOI] [PubMed] [Google Scholar]
  15. Yamashita T., Arens M., Green M. Adenovirus deoxyribonucleic acid replication. II. Synthesis of viral deoxyribonucleic acid in vitro by a nuclear membrane fraction from infected KB cells. J Biol Chem. 1975 May 10;250(9):3273–3279. [PubMed] [Google Scholar]
  16. Yamashita T., Shimojo H. Replication of adenovirus 12 deoxyribonucleic acid in association with the nuclear membrane. Jpn J Microbiol. 1973 Sep;17(5):419–423. doi: 10.1111/j.1348-0421.1973.tb00793.x. [DOI] [PubMed] [Google Scholar]
  17. van der Vliet P. C., Sussenbach J. S. The mechanism of adenovirus-DNA synthesis in isolated nuclei. Eur J Biochem. 1972 Nov 7;30(3):584–592. doi: 10.1111/j.1432-1033.1972.tb02130.x. [DOI] [PubMed] [Google Scholar]

Articles from Journal of Virology are provided here courtesy of American Society for Microbiology (ASM)

RESOURCES