Skip to main content
Nucleic Acids Research logoLink to Nucleic Acids Research
. 1980 Sep 11;8(17):3993–4007. doi: 10.1093/nar/8.17.3993

Differential effect of aphidicolin on adenovirus DNA synthesis and cellular DNA synthesis.

M M Kwant, P C van der Vliet
PMCID: PMC324210  PMID: 6777759

Abstract

There is strong evidence for a participation of DNA polymerase gamma in the replication of adenovirus (Ad) DNA. To study a possible additional role of DNA polymerase alpha we measured the effect of aphidicolin on viral DNA replication. In intact cells, aphidicolin inhibits Ad DNA synthesis weakly. The drug concentration required for 50% inhibition of Ad DNA replication was 300-400 fold higher than for a similar effect on cellular DNA synthesis. Such a differential inhibition was also observed in AGMK cells doubly infected with SV40 and the simian adenovirus SA7. No evidence was found for modification of aphidicolin in infected cells or for a change in aphidicolin sensitivity of DNA polymerase alpha after infection. The extent of inhibition of purified DNA polymerase alpha was dependent upon the dCTP concentration. The same situation was observed when DNA synthesis was studied in isolated nuclei from uninfected cells. However, in nuclei from Ad infected cells no effect of dCTP on aphidicolin sensitivity was found. These results were taken as evidence that DNA polymerase alpha does not participate in the replication of adenovirus DNA.

Full text

PDF
3993

Selected References

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

  1. Arens M., Yamashita T., Padmanabhan R., Tsuruo T., Green M. Adenovirus deoxyribonucleic acid replication. Characterization of the enzyme activities of a soluble replication system. J Biol Chem. 1977 Nov 25;252(22):7947–7954. [PubMed] [Google Scholar]
  2. Brison O., Kedinger C., Wilhelm J. Enzymatic properties of viral replication complexes isolated from adenovirus type 2-infected HeLa cell nuclei. J Virol. 1977 Nov;24(2):423–435. doi: 10.1128/jvi.24.2.423-435.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Bucknall R. A., Moores H., Simms R., Hesp B. Antiviral effects of aphidicolin, a new antibiotic produced by Cephalosporium aphidicola. Antimicrob Agents Chemother. 1973 Sep;4(3):294–298. doi: 10.1128/aac.4.3.294. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Challberg M. D., Kelly T. J., Jr Adenovirus DNA replication in vitro. Proc Natl Acad Sci U S A. 1979 Feb;76(2):655–659. doi: 10.1073/pnas.76.2.655. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Fraser J. M., Huberman J. A. In vitro HeLa cell DNA synthesis. II. Partial characterization of soluble factors stimulating nuclear dna synthesis. Biochim Biophys Acta. 1978 Sep 27;520(2):271–284. doi: 10.1016/0005-2787(78)90226-5. [DOI] [PubMed] [Google Scholar]
  6. Habara A., Kano K., Nagano H., Mano Y., Ikegami S., Yamashita T. Inhibition of DNA synthesis in the adenovirus DNA replication complex by aphidicolin and 2',3'-dideoxythymidine triphosphate. Biochem Biophys Res Commun. 1980 Jan 15;92(1):8–12. doi: 10.1016/0006-291x(80)91511-9. [DOI] [PubMed] [Google Scholar]
  7. Hanaoka F., Kato H., Ikegami S., Oashi M., Yamada M. Aphidicolin does inhibit repair replication in HeLa cells. Biochem Biophys Res Commun. 1979 Mar 30;87(2):575–580. doi: 10.1016/0006-291x(79)91833-3. [DOI] [PubMed] [Google Scholar]
  8. Horwitz M. S., Brayton C., Baum S. G. Synthesis of type 2 adenovirus DNA in the presence of cycloheximide. J Virol. 1973 Apr;11(4):544–551. doi: 10.1128/jvi.11.4.544-551.1973. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Ikegami S., Taguchi T., Ohashi M., Oguro M., Nagano H., Mano Y. Aphidicolin prevents mitotic cell division by interfering with the activity of DNA polymerase-alpha. Nature. 1978 Oct 5;275(5679):458–460. doi: 10.1038/275458a0. [DOI] [PubMed] [Google Scholar]
  10. Kedinger C., Brison O., Perrin F., Wilhelm J. Structural analysis of viral replicative intermediates isolated from adenovirus type 2-infected HeLa cell nuclei. J Virol. 1978 May;26(2):364–379. doi: 10.1128/jvi.26.2.364-379.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Knopf K. W., Yamada M., Weissbach A. HeLa cell DNA polymerase gamma: further purification and properties of the enzyme. Biochemistry. 1976 Oct 5;15(20):4540–4548. doi: 10.1021/bi00665a032. [DOI] [PubMed] [Google Scholar]
  12. Krokan H., Schaffer P., DePamphilis M. L. Involvement of eucaryotic deoxyribonucleic acid polymerases alpha and gamma in the replication of cellular and viral deoxyribonucleic acid. Biochemistry. 1979 Oct 2;18(20):4431–4443. doi: 10.1021/bi00587a025. [DOI] [PubMed] [Google Scholar]
  13. Levine A. J., Van Der Vliet P. C., Rosenwirth B., Rabek J., Frenkel G., Ensinger M. Adenovirus-infected, cell-specific, DNA-binding proteins. Cold Spring Harb Symp Quant Biol. 1975;39(Pt 1):559–566. doi: 10.1101/sqb.1974.039.01.069. [DOI] [PubMed] [Google Scholar]
  14. 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]
  15. Longiaru M., Ikeda J. E., Jarkovsky Z., Horwitz S. B., Horwitz M. S. The effect of aphidicolin on adenovirus DNA synthesis. Nucleic Acids Res. 1979 Jul 25;6(10):3369–3386. doi: 10.1093/nar/6.10.3369. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Longiaru M., Ikeda J. E., Jarkovsky Z., Horwitz S. B., Horwitz M. S. The effect of aphidicolin on adenovirus DNA synthesis. Nucleic Acids Res. 1979 Jul 25;6(10):3369–3386. doi: 10.1093/nar/6.10.3369. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Oguro M., Suzuki-Hori C., Nagano H., Mano Y., Ikegami S. The mode of inhibitory action by aphidicolin on eukaryotic DNA polymerase alpha. Eur J Biochem. 1979 Jul;97(2):603–607. doi: 10.1111/j.1432-1033.1979.tb13149.x. [DOI] [PubMed] [Google Scholar]
  18. Ohashi M., Taguchi T., Ikegami S. Aphidicolin: a specific inhibitor of DNA polymerases in the cytosol of rat liver. Biochem Biophys Res Commun. 1978 Jun 29;82(4):1084–1090. doi: 10.1016/0006-291x(78)90298-x. [DOI] [PubMed] [Google Scholar]
  19. Pedrali-Noy G., Spadari S. Effect of aphidicolin on viral and human DNA polymerases. Biochem Biophys Res Commun. 1979 Jun 27;88(4):1194–1202. doi: 10.1016/0006-291x(79)91106-9. [DOI] [PubMed] [Google Scholar]
  20. Smith R. G., Abrell J. W., Lewis B. J., Gallo R. C. Serological analysis of human deoxyribonucleic acid polymerases. Preparation and properties of antiserum to deoxyribonucleic acid polymerase I from human lymphoid cells. J Biol Chem. 1975 Mar 10;250(5):1702–1709. [PubMed] [Google Scholar]
  21. Tate V. E., Philipson L. Parental adenovirus DNA accumulates in nucleosome-like structures in infected cells. Nucleic Acids Res. 1979 Jun 25;6(8):2769–2785. doi: 10.1093/nar/6.8.2769. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Van der Vliet P. C., Zandberg J., Jansz H. S. Evidence for a function of the adenovirus DNA-binding protein in initiation in DNA synthesis as well as in elongation of nascent DNA chains. Virology. 1977 Jul 1;80(1):98–110. doi: 10.1016/0042-6822(77)90383-x. [DOI] [PubMed] [Google Scholar]
  23. Weissbach A. The functional roles of mammalian DNA polymerase. Arch Biochem Biophys. 1979 Dec;198(2):386–396. doi: 10.1016/0003-9861(79)90511-3. [DOI] [PubMed] [Google Scholar]
  24. Winnacker E. L. Adenovirus DNA: structure and function of a novel replicon. Cell. 1978 Aug;14(4):761–773. doi: 10.1016/0092-8674(78)90332-x. [DOI] [PubMed] [Google Scholar]
  25. Wist E., Prydz H. The effect of aphidicolin on DNA synthesis in isolated HeLa cell nuclei. Nucleic Acids Res. 1979 Apr;6(4):1583–1590. doi: 10.1093/nar/6.4.1583. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. de Jong A., van der Vliet P. C., Jansz H. S. DNA polymerases in adenovirus type 5-infected and uninfected KB cells. Induction of an alpha-type DNA polymerase in adenovirus type 5-infected and in fast growing cells. Biochim Biophys Acta. 1977 May 17;476(2):156–165. doi: 10.1016/0005-2787(77)90092-2. [DOI] [PubMed] [Google Scholar]
  27. van der Vliet P. C., Kwant M. M. Role of DNA polymerase gamma in adenovirus DNA replication. Nature. 1978 Nov 30;276(5687):532–534. doi: 10.1038/276532a0. [DOI] [PubMed] [Google Scholar]
  28. 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 Nucleic Acids Research are provided here courtesy of Oxford University Press

RESOURCES