1887

Abstract

We report the covalent addition of [P]dCMP to a protein from group A adenovirus 12 (Ad12)-infected human (KB) cells , using crude extracts. Synthesis of the 60K protein-dCMP complex required a DNA template containing a terminally located adenovirus replication origin; the protein-dCMP bond was alkali-labile but acid-stable. We therefore conclude that this product is the Ad12 terminal protein precursor (pTP)-dCMP initiation complex for DNA replication. Synthesis of Ad12 pTP-dCMP was specific for dCTP but was stimulated by dATP. In contrast to Ad2, the Ad12 initiation reaction required ATP. Antipeptide antiserum targeted to Ad DNA polymerase inhibited Ad12 pTP-dCMP synthesis , providing evidence that Ad DNA polymerase catalyses dCMP addition to pTP during initiation.

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/content/journal/jgv/10.1099/0022-1317-72-2-427
1991-02-01
2021-10-22
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References

  1. Challberg M. D., Kelly T. J. Jr 1979; Adenovirus DNA replication in vitro . Proceedings of the National Academy of Sciences, U,. S,. A. 76:655–659
    [Google Scholar]
  2. Challberg M. D., Kelly T. J. Jr 1989; Animal virus DNA replication. Annual Review of Biochemistry 58:671–717
    [Google Scholar]
  3. Chen M., Horwitz M. S. 1989; Dissection of functional domains of adenovirus DNA polymerase by linker-insertion mutagenesis. Proceedings of the National Academy of Sciences, U,. S,. A 86:6116–6120
    [Google Scholar]
  4. Chowrira B. M., Lucher L. A. 1990; Extracts of hamster cells abortively infected with human adenovirus type 12 are competent to support initiation of viral DNA replication. Virology 176:289–291
    [Google Scholar]
  5. Enomoto T., Lichy J. H., Ikeda J.-E., Hurwitz J. 1981; Adenovirus DNA replication in vitro: purification of the terminal protein in a functional form. Proceedings of the National Academy of Sciences, U,. S,. A. 78:6779–6783
    [Google Scholar]
  6. Esche H., Siegmann B. 1982; Expression of early viral gene products in adenovirus type 12-infected and transformed cells. Journal of General Virology 60:99–113
    [Google Scholar]
  7. Hay R. T. 1985; Origin of adenovirus DNA replication: role of the nuclear factor I binding site in vivo . Journal of Molecular Biology 186:129–136
    [Google Scholar]
  8. Kelly T. J. Jr, Wold M. S., Li J. 1988; Initiation of viral DNA replication. Advances in Virus Research 34:1–42
    [Google Scholar]
  9. Kenny M. K., Hurwitz J. 1988; Initiation of adenovirus DNA replication. II. Structural requirements using synthetic oligonucleotide adenovirus templates. Journal of Biological Chemistry 263:9809–9817
    [Google Scholar]
  10. Khuntirat B., Lucher L. A. 1990; A microscale analytical batch chromatographic method for detecting soluble viral DNA-binding proteins in crude extracts. Journal of Virological Methods 29:97–104
    [Google Scholar]
  11. Lichy J. H., Horwitz M. S., Hurwitz J. 1981; Formation of a covalent complex between the 80, 000-dalton adenovirus terminal protein and 5′-dCMP in vitro . Proceedings of the National Academy of Sciences, U,. S,. A. 78:2678–2682
    [Google Scholar]
  12. Lucher L. A. 1990; Adenovirus type 12 tumour antigen synthesis differs during infection of permissive and non-permissive cells. Journal of General Virology 71:579–583
    [Google Scholar]
  13. Shiroki K., Shimojo H., Yamaguchi K. 1974; The viral DNA replication complex of adenovirus 12. Virology 60:192–199
    [Google Scholar]
  14. Shiroki K., Hashimoto S., Saito I., Fukui Y., Fukui Y., Kato H., Shimojo H. 1984; Expression of the E4 gene is required for establishment of soft-agar colony-forming rat cell lines transformed by the adenovirus 12 E1 gene. Journal of Virology 50:854–863
    [Google Scholar]
  15. Shu L., Hong J. S., Wei Y., Engler J. A. 1986; Nucleotide sequence of the genes encoded in early region 2b of human adenovirus type 12. Gene 46:187–195
    [Google Scholar]
  16. Stillman B. W., Tamanoi F., Mathews M. B. 1982; Purification of an adenovirus-coded DNA polymerase that is required for initiation of DNA replication. Cell 31:613–623
    [Google Scholar]
  17. Sussenbach J. S., Van der Vliet P. C. 1983; The mechanism of adenovirus DNA replication and the characterization of replication proteins. Current Topics in Microbiology and Immunology 109:53–73
    [Google Scholar]
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