Skip to main content
NCBI home page
Journal of Virology logoLink to Journal of Virology
. 1977 May;22(2):402–411. doi: 10.1128/jvi.22.2.402-411.1977

Adenovirus type 2 coded single-stranded DNA binding protein: in vivo phosphorylation and modification.

Y H Jeng, W S Wold, K Sugawara, Z Gilead, M Green
PMCID: PMC515731  PMID: 864833

Abstract

The adenovirus type 2-coded single-stranded DNA binding protein (DBP) was shown to be a phosphoprotein and to exist in at least two forms that differ in mobility by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. After a 30-min pulse with [35S]methionine or 32PO4, 35S- or 32P-labeled DBP had a nominal molecular weight of 74,000 whereas after a 30-min label followed by a 20-h chase, 35S- and 32P-labeled DBP had a nominal molecular weight of 77,000. Both large and small forms of 35S- and 32P-labeled DBP bound to single-stranded DNA-cellulose columns and were eluted by 0.4 to 0.6 M NaCl; both forms also were immunoprecipitated by antiserum against adenovirus type 1-simian virus 40-induced tumor cells (this antiserum contains antibodies against DBP) and by monospecific antiserum against 95 to 99% purified DBP. With highly purified 32P-DBP labeled 7 to 10 h postinfection, it was shown that the 32P radioactivity was firmly associated with protein material (i.e., not contaminating nucleic acids or phospholipids) and had properties expected of a phosphoester of an amino acid; paper electrophoresis of acid hydrolysates of this preparation identified phosphoserine but not phosphothreonine. Phosphoserine but not phosphothreonine was also identified in acid hydrolysates of another preparation of 32P-DBP labeled for 30 min, chased for 20 h, and then immunoprecipitated by adenovirus type 1-simian virus 40 antiserum.

Full text

PDF
402

Images in this article

405Image
on p.405
406Image
on p.406
407Image
on p.407
407Image
on p.407
409Image
on p.409
409Image
on p.409

Selected References

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

  1. Alberts B. M., Frey L. T4 bacteriophage gene 32: a structural protein in the replication and recombination of DNA. Nature. 1970 Sep 26;227(5265):1313–1318. doi: 10.1038/2271313a0. [DOI] [PubMed] [Google Scholar]
  2. Allerton S. E., Perlmann G. E. Chemical characterization of the phosphoprotein phosvitin. J Biol Chem. 1965 Oct;240(10):3892–3898. [PubMed] [Google Scholar]
  3. Anderson C. W., Baum P. R., Gesteland R. F. Processing of adenovirus 2-induced proteins. J Virol. 1973 Aug;12(2):241–252. doi: 10.1128/jvi.12.2.241-252.1973. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Cantoni G. L. Biological methylation: selected aspects. Annu Rev Biochem. 1975;44:435–451. doi: 10.1146/annurev.bi.44.070175.002251. [DOI] [PubMed] [Google Scholar]
  5. GREEN M., PINA M. Biochemical studies on adenovirus multiplication. IV. Isolation, purification, and chemical analysis of adenovirus. Virology. 1963 May;20:199–207. doi: 10.1016/0042-6822(63)90157-0. [DOI] [PubMed] [Google Scholar]
  6. Gielkens A. L., Van Zaane D., Bloemers H. P., Bloemendal H. Synthesis of Rauscher murine leukemia virus-specific polypeptides in vitro. Proc Natl Acad Sci U S A. 1976 Feb;73(2):356–360. doi: 10.1073/pnas.73.2.356. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Gilden R. V., Kern J., Freeman A. E., Martin C. E., McAllister R. C., Turner H. C., Huebner R. J. T and tumour antigens of adenovirus group C-infected and transformed cells. Nature. 1968 Aug 3;219(5153):517–518. doi: 10.1038/219517a0. [DOI] [PubMed] [Google Scholar]
  8. Gilead Z., Arens M. Q., Bhaduri S., Shanmugam G., Green M. Tumour antigen specificity of a DNA-binding protein from cells infected with adenovirus 2. Nature. 1975 Apr 10;254(5500):533–536. doi: 10.1038/254533a0. [DOI] [PubMed] [Google Scholar]
  9. 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]
  10. Gumport R. I., Lehman I. R. Structure of the DNA ligase-adenylate intermediate: lysine (epsilon-amino)-linked adenosine monophosphoramidate. Proc Natl Acad Sci U S A. 1971 Oct;68(10):2559–2563. doi: 10.1073/pnas.68.10.2559. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Harter M. L., Shanmugam G., Wold W. S., Green M. Detection of adenovirus type 2-induced early polypeptides using cycloheximide pretreatment to enhance viral protein synthesis. J Virol. 1976 Jul;19(1):232–242. doi: 10.1128/jvi.19.1.232-242.1976. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Kabat D. Phosphorylation of ribosomal proteins in rabbit reticulocytes. A cell-free system with ribosomal protein kinase activity. Biochemistry. 1971 Jan 19;10(2):197–203. doi: 10.1021/bi00778a001. [DOI] [PubMed] [Google Scholar]
  13. Kleinsmith L. J., Stein J., Stein G. Dephosphorylation of nonhistone proteins specifically alters the pattern of gene transcription in reconstituted chromatin. Proc Natl Acad Sci U S A. 1976 Apr;73(4):1174–1178. doi: 10.1073/pnas.73.4.1174. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
  15. 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]
  16. 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]
  17. PERLMANN G. E. The nature of phosphorus linkages in phosphoproteins. Adv Protein Chem. 1955;10:1–30. doi: 10.1016/s0065-3233(08)60103-5. [DOI] [PubMed] [Google Scholar]
  18. Philipson L., Pettersson U., Lindberg U. Molecular biology of adenoviruses. Virol Monogr. 1975;14:1–115. doi: 10.1007/978-3-7091-8391-5_1. [DOI] [PubMed] [Google Scholar]
  19. Rahmsdorf H. J., Pai S. H., Ponta H., Herrlich P., Roskoski R., Jr, Schweiger M., Studier F. W. Protein kinase induction in Escherichia coli by bacteriophage T7. Proc Natl Acad Sci U S A. 1974 Feb;71(2):586–589. doi: 10.1073/pnas.71.2.586. [DOI] [PMC free article] [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. Rubin C. S., Rosen O. M. Protein phosphorylation. Annu Rev Biochem. 1975;44:831–887. doi: 10.1146/annurev.bi.44.070175.004151. [DOI] [PubMed] [Google Scholar]
  23. Sambrook J., Botchan M., Gallimore P., Ozanne B., Pettersson U., Williams J., Sharp P. A. Viral DNA sequences in cells transformed by simian virus 40, adenovirus type 2 and adenovirus type 5. Cold Spring Harb Symp Quant Biol. 1975;39(Pt 1):615–632. doi: 10.1101/sqb.1974.039.01.075. [DOI] [PubMed] [Google Scholar]
  24. Shanmugam G., Bhaduri S., Arens M., Green M. DNA binding proteins in the cytoplasm and in a nuclear membrane complex isolated from uninfected and adenovirus 2 infected cells. Biochemistry. 1975 Jan 28;14(2):332–337. doi: 10.1021/bi00673a020. [DOI] [PubMed] [Google Scholar]
  25. Stein G. S., Spelsberg T. C., Kleinsmith L. J. Nonhistone chromosomal proteins and gene regulation. Science. 1974 Mar 1;183(4127):817–824. doi: 10.1126/science.183.4127.817. [DOI] [PubMed] [Google Scholar]
  26. Studier F. W. Analysis of bacteriophage T7 early RNAs and proteins on slab gels. J Mol Biol. 1973 Sep 15;79(2):237–248. doi: 10.1016/0022-2836(73)90003-x. [DOI] [PubMed] [Google Scholar]
  27. 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]
  28. Sugawara K., Gilead Z., Wold W. S., Green M. Immunofluorescence study of the adenovirus type 2 single-stranded DNA binding protein in infected and transformed cells. J Virol. 1977 May;22(2):527–539. doi: 10.1128/jvi.22.2.527-539.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. 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]
  30. 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]
  31. Walter G., Martin H. Simian virus 40-specific proteins in HeLa cells infected with nondefective adenovirus 2-simian virus 40 hybrid viruses. J Virol. 1975 Nov;16(5):1236–1247. doi: 10.1128/jvi.16.5.1236-1247.1975. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Weber J. Genetic analysis of adenovirus type 2 III. Temperature sensitivity of processing viral proteins. J Virol. 1976 Feb;17(2):462–471. doi: 10.1128/jvi.17.2.462-471.1976. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. 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]
  34. Yamashita T., Green M. Adenovirus DNA replication. I. Requirement for protein synthesis and isolation of nuclear membrane fractions containing newly synthesized viral DNA and proteins. J Virol. 1974 Sep;14(3):412–420. doi: 10.1128/jvi.14.3.412-420.1974. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. 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]

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

RESOURCES