Skip to main content
Proceedings of the National Academy of Sciences of the United States of America logoLink to Proceedings of the National Academy of Sciences of the United States of America
. 1981 May;78(5):2678–2682. doi: 10.1073/pnas.78.5.2678

Formation of a covalent complex between the 80,000-dalton adenovirus terminal protein and 5'-dCMP in vitro.

J H Lichy, M S Horwitz, J Hurwitz
PMCID: PMC319420  PMID: 6942401

Abstract

An in vitro adenovirus DNA replication system catalyzed the formation of a covalent complex between an 80,000-dalton protein and 5'-dCMP in the presence of [alpha-32P-dCTP, MgCl2, ATP, and adenovirus (Ad) DNA with a protein covalently bound to the 5' end of each strand (Ad DNA-prot). The requirement for Ad DNA-prot in this reaction was similar to that for in vitro DNA replication. When dATP, dTTP, and the 2',3'-dideoxynucleoside triphosphate (ddNTP) ddGTP were included in the reaction mixture, an elongated complex was detected, which consisted of an 80,000-dalton protein bound to a 26-base oligonucleotide. Formation of the elongated product, but not of the protein-dCMP complex, was inhibited by ddATP, ddCTP, or ddTTP. The requirements for formation of the protein-dCMP complex, the nature of the linkage between protein and dCMP, the size of the protein, and the existence of elongated forms indicated that the protein associated with the complex was identical to the 80,000-dalton Ad terminal protein found on replicating DNA molecules as described by Challberg et al. [Challberg, M. D., Desiderio, S. V. & Kelly, T. J., Jr. (1980) Proc. Natl. Acad. Sci. USA 77, 5105-5109].

Full text

PDF
2679

Images in this article

Selected References

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

  1. Arrand J. R., Roberts R. J. The nucleotide sequences at the termini of adenovirus-2 DNA. J Mol Biol. 1979 Mar 15;128(4):577–594. doi: 10.1016/0022-2836(79)90294-8. [DOI] [PubMed] [Google Scholar]
  2. Carusi E. A. Evidence for blocked 5'-termini in human adenovirus DNA. Virology. 1977 Jan;76(1):380–394. doi: 10.1016/0042-6822(77)90310-5. [DOI] [PubMed] [Google Scholar]
  3. Challberg M. D., Desiderio S. V., Kelly T. J., Jr Adenovirus DNA replication in vitro: characterization of a protein covalently linked to nascent DNA strands. Proc Natl Acad Sci U S A. 1980 Sep;77(9):5105–5109. doi: 10.1073/pnas.77.9.5105. [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. Challberg M. D., Kelly T. J., Jr Adenovirus DNA replication in vitro: origin and direction of daughter strand synthesis. J Mol Biol. 1979 Dec 25;135(4):999–1012. doi: 10.1016/0022-2836(79)90524-2. [DOI] [PubMed] [Google Scholar]
  6. Horwitz M. S., Ariga H. Multiple rounds of adenovirus DNA synthesis in vitro. Proc Natl Acad Sci U S A. 1981 Mar;78(3):1476–1480. doi: 10.1073/pnas.78.3.1476. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Horwitz M. S., Kaplan L. M., Abboud M., Maritato J., Chow L. T., Broker T. R. Adenovirus DNA replication in soluble extracts of infected cell nuclei. Cold Spring Harb Symp Quant Biol. 1979;43(Pt 2):769–780. doi: 10.1101/sqb.1979.043.01.084. [DOI] [PubMed] [Google Scholar]
  8. Ikeda J. E., Enomoto T., Hurwitz J. Replication of adenovirus DNA-protein complex with purified proteins. Proc Natl Acad Sci U S A. 1981 Feb;78(2):884–888. doi: 10.1073/pnas.78.2.884. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Ikeda J. E., Longiaru M., Horwitz M. S., Hurwitz J. Elongation of primed DNA templates by eukaryotic DNA polymerases. Proc Natl Acad Sci U S A. 1980 Oct;77(10):5827–5831. doi: 10.1073/pnas.77.10.5827. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Kaplan L. M., Ariga H., Hurwitz J., Horwitz M. S. Complementation of the temperature-sensitive defect in H5ts125 adenovirus DNA replication in vitro. Proc Natl Acad Sci U S A. 1979 Nov;76(11):5534–5538. doi: 10.1073/pnas.76.11.5534. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. 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]
  12. Rekosh D. M., Russell W. C., Bellet A. J., Robinson A. J. Identification of a protein linked to the ends of adenovirus DNA. Cell. 1977 Jun;11(2):283–295. doi: 10.1016/0092-8674(77)90045-9. [DOI] [PubMed] [Google Scholar]
  13. Robinson A. J., Bellett J. D. A circular DNA-protein complex adenoviruses and its possible role in DNA replication. Cold Spring Harb Symp Quant Biol. 1975;39(Pt 1):523–531. doi: 10.1101/sqb.1974.039.01.064. [DOI] [PubMed] [Google Scholar]
  14. Robinson A. J., Bodnar J. W., Coombs D. H., Pearson G. D. Replicating adenovirus 2 DNA molecules contain terminal protein. Virology. 1979 Jul 15;96(1):143–158. doi: 10.1016/0042-6822(79)90180-6. [DOI] [PubMed] [Google Scholar]
  15. Robinson A. J., Younghusband H. B., Bellett A. J. A circula DNA-protein complex from adenoviruses. Virology. 1973 Nov;56(1):54–69. doi: 10.1016/0042-6822(73)90287-0. [DOI] [PubMed] [Google Scholar]
  16. Roninson I., Padmanabhan R. Studies on the nature of the linkage between the terminal protein and the adenovirus DNA. Biochem Biophys Res Commun. 1980 May 14;94(1):398–405. doi: 10.1016/s0006-291x(80)80234-8. [DOI] [PubMed] [Google Scholar]
  17. Shinagawa M., Padmanabhan R. Comparative sequence analysis of the inverted terminal repetitions from different adenoviruses. Proc Natl Acad Sci U S A. 1980 Jul;77(7):3831–3835. doi: 10.1073/pnas.77.7.3831. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Shinagawa M., Padmanabhan R. Nucleotide sequence at the inverted terminal repetition of adenovirus type 2 DNA. Biochem Biophys Res Commun. 1979 Apr 13;87(3):671–678. doi: 10.1016/0006-291x(79)92011-4. [DOI] [PubMed] [Google Scholar]
  19. Stillman B. W., Bellett A. J. An adenovirus protein associated with the ends of replicating DNA molecules. Virology. 1979 Feb;93(1):69–79. doi: 10.1016/0042-6822(79)90276-9. [DOI] [PubMed] [Google Scholar]
  20. Stillman B. W., Lewis J. B., Chow L. T., Mathews M. B., Smart J. E. Identification of the gene and mRNA for the adenovirus terminal protein precursor. Cell. 1981 Feb;23(2):497–508. doi: 10.1016/0092-8674(81)90145-8. [DOI] [PubMed] [Google Scholar]

Articles from Proceedings of the National Academy of Sciences of the United States of America are provided here courtesy of National Academy of Sciences

RESOURCES