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. 1980 Aug;35(2):314–319. doi: 10.1128/jvi.35.2.314-319.1980

Adenovirus-specific DNA-binding protein inhibits the hydrolysis of DNA by DNase in vitro.

K Nass, G D Frenkel
PMCID: PMC288815  PMID: 6893723

Abstract

The adenovirus-specific DNA-binding protein was isolated from adenovirus type 5-infected KB cells and shown to possess DNase inhibitor activity. The protein decreased the rate of hydrolysis of single-strand DNA proportionately to its concentration in the reaction. Two peaks of activity were obtained upon sedimentation in a glycerol gradient, probably corresponding to the two major adenovirus-specific polypeptides in the preparation (molecular weights, 72,000 and 44,000). The DNase inhibitor activity of the adenovirus DNA-binding protein was distinguishable from that of the cellular DNA-binding protein, which we have described previously (K, Nass and G. D. Frenkel, J. Biol. Chem. 254:3407-3410, 1979), by its pattern of sedimentation and by the effect of temperature on the two activities. For the adenovirus DNA-binding protein, the ratio of DNase inhibitor activity at 43 degrees C to that at 30 degrees C was approximately 14, whereas for the cellular protein this ratio was less than 3. The DNase inhibitor activity with the temperature coefficient of 14 was absent from cells infected with adenovirus type 5 ts125 at 40 degrees C. DNase inhibition is a simple, sensitive, quantitative method for assay of the adenovirus DNA-binding protein.

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Selected References

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  1. Axelrod N. Phosphoproteins of adenovirus 2. Virology. 1978 Jun 15;87(2):366–383. doi: 10.1016/0042-6822(78)90141-1. [DOI] [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. Gilead Z., Sugawara K., Shanmugam G., Green M. Synthesis of the adenovirus-coded DNA binding protein in infected cells. J Virol. 1976 May;18(2):454–460. doi: 10.1128/jvi.18.2.454-460.1976. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Ginsberg H. S., Lundholm U., Linné T. Adenovirus DNA-binding protein in cells infected with wild-type 5 adenovirus and two DNA-minus, temperature-sensitive mutants, H5ts125 and H5ts149. J Virol. 1977 Jul;23(1):142–151. doi: 10.1128/jvi.23.1.142-151.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Grodzicker T., Anderson C., Sambrook J., Mathews M. B. The physical locations of structural genes in adenovirus DNA. Virology. 1977 Jul 1;80(1):111–126. doi: 10.1016/0042-6822(77)90384-1. [DOI] [PubMed] [Google Scholar]
  6. 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]
  7. Horwitz M. S. Temperature-sensitive replication of H5ts125 adenovirus DNA in vitro. Proc Natl Acad Sci U S A. 1978 Sep;75(9):4291–4295. doi: 10.1073/pnas.75.9.4291. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Jeng Y. H., Wold W. S., Sugawara K., Gilead Z., Green M. Adenovirus type 2 coded single-stranded DNA binding protein: in vivo phosphorylation and modification. J Virol. 1977 May;22(2):402–411. doi: 10.1128/jvi.22.2.402-411.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. 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]
  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. Levinson A. D., Postel E. H., Levine A. J. In vivo and in vitro phosphorylation of the adenovirus type 5 single strand-specific DNA-binding protein. Virology. 1977 Jun 1;79(1):144–159. doi: 10.1016/0042-6822(77)90341-5. [DOI] [PubMed] [Google Scholar]
  12. Levinson A., Levine A. J., Anderson S., Osborn M., Rosenwirth B., Weber K. The relationship between group C adenovirus tumor antigen and the adenovirus single-strand DNA-binding protein. Cell. 1976 Apr;7(4):575–584. doi: 10.1016/0092-8674(76)90208-7. [DOI] [PubMed] [Google Scholar]
  13. Lewis J. B., Atkins J. F., Baum P. R., Solem R., Gesteland R. F., Anderson C. W. Location and identification of the genes for adenovirus type 2 early polypeptides. Cell. 1976 Jan;7(1):141–151. doi: 10.1016/0092-8674(76)90264-6. [DOI] [PubMed] [Google Scholar]
  14. Linné T., Jörnvall H., Philipson L. Purification and characterization of the phosphorylated DNA-binding protein from adenovirus-type-2-infected cells. Eur J Biochem. 1977 Jun 15;76(2):481–490. doi: 10.1111/j.1432-1033.1977.tb11618.x. [DOI] [PubMed] [Google Scholar]
  15. Mackay V., Linn S. Selective inhibition of the dnase activity of the recBC enzyme by the DNA binding protein from Escherichia coli. J Biol Chem. 1976 Jun 25;251(12):3716–3719. [PubMed] [Google Scholar]
  16. Molineux I. J., Gefter M. L. Properties of the Escherichia coli DNA-binding (unwinding) protein interaction with nucleolytic enzymes and DNA. J Mol Biol. 1975 Nov 15;98(4):811–825. doi: 10.1016/s0022-2836(75)80012-x. [DOI] [PubMed] [Google Scholar]
  17. Naas K., Frenkel G. D. A deoxyribonucleic acid binding protein from KB cells which inhibits deoxyribonuclease activity on single-stranded DNA. J Biol Chem. 1979 May 10;254(9):3407–3410. [PubMed] [Google Scholar]
  18. Nass K., Frenkel G. D. Adenovirus-induced inhibition of cellular DNase. J Virol. 1978 May;26(2):540–543. doi: 10.1128/jvi.26.2.540-543.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Robin J., Bourgaux-Ramoisy D., Bourgaux P. Single-stranded regions in replicating DNA of adenovirus type 2. J Gen Virol. 1973 Aug;20(2):233–237. doi: 10.1099/0022-1317-20-2-233. [DOI] [PubMed] [Google Scholar]
  20. Rosenwirth B., Anderson C., Levine A. J. Tryptic fingerprint analysis of adenovirus types 2, 5 and 12 DNA-Binding proteins. Virology. 1976 Feb;69(2):617–625. doi: 10.1016/0042-6822(76)90490-6. [DOI] [PubMed] [Google Scholar]
  21. Rosenwirth B., Shiroki K., Levine A. J., Shimojo H. Isolation and characterization of adenovirus type 12 DNA binding proteins. Virology. 1975 Sep;67(1):14–23. doi: 10.1016/0042-6822(75)90399-2. [DOI] [PubMed] [Google Scholar]
  22. Russell W. C., Blair G. E. Polypeptide phosphorylation in adenovirus-infected cells. J Gen Virol. 1977 Jan;34(1):19–35. doi: 10.1099/0022-1317-34-1-19. [DOI] [PubMed] [Google Scholar]
  23. Sugawara K., Gilead Z., Green M. Purification and molecular characterization of adenovirus type 2 DNA-binding protein. J Virol. 1977 Jan;21(1):338–346. doi: 10.1128/jvi.21.1.338-346.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Sussenbach J. S., van der Vliet P. C., Ellens D. J., Jansz H. S. Linear intermediates in the replication of adenovirus DNA. Nat New Biol. 1972 Sep 13;239(89):47–49. [PubMed] [Google Scholar]
  25. 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]
  26. 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]
  27. 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]
  28. van der Eb A. J. Intermediates in type 5 adenovirus DNA replication. Virology. 1973 Jan;51(1):11–23. doi: 10.1016/0042-6822(73)90361-9. [DOI] [PubMed] [Google Scholar]
  29. van der Vliet P. C., Keegstra W., Jansz H. S. Complex formation between the adenovirus type 5 DNA-binding protein and single-stranded DNA. Eur J Biochem. 1978 May 16;86(2):389–398. doi: 10.1111/j.1432-1033.1978.tb12321.x. [DOI] [PubMed] [Google Scholar]
  30. van der Vliet P. C., Levine A. J. DNA-binding proteins specific for cells infected by adenovirus. Nat New Biol. 1973 Dec 12;246(154):170–174. doi: 10.1038/newbio246170a0. [DOI] [PubMed] [Google Scholar]

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