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. 1979 Mar;29(3):938–948. doi: 10.1128/jvi.29.3.938-948.1979

Purification and characterization of an early glycoprotein from adenovirus type 2-infected cells.

H Persson, C Signäs, L Philipson
PMCID: PMC353253  PMID: 448802

Abstract

An adenovirus type 2 early glycoprotein with an apparent molecular weight of 19,000 (E19K) in sodium dodecyl sulfate-polyacrylamide gels has been extensively purified. Purification involved detergent solubilization of membrane fractions from infected cells, followed by affinity chromatography on a lectin column and DEAE-Sephadex chromatography. The purified material contained three polypeptides (E40K, E19K, E17.5K), with approximately 90% of the material in the E19K moiety. All three polypeptides yielded identical tryptic peptide maps. The E19K polypeptide contained glucosamine as revealed by [3H]glucosamine labeling of infected cells and amino acid analysis of the purified protein. Immunoprecipitation with a monospecific antiserum showed that the E19K polypeptide started to be synthesized at 2 h, with a maximal rate at 4 h after infection. It was also synthesized at a low rate late in the infectious cycle (12 to 24 h postinfection). Immunoprecipitation from three adenovirus type 2-transformed hamster embryo cell lines and two adenovirus type 2-transformed rat cell lines revealed that one of the hamster cell lines (ad2HE4) and one of the rat cell lines (A2T2C4) expressed this protein.

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Selected References

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

  1. 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]
  2. Berk A. J., Sharp P. A. Structure of the adenovirus 2 early mRNAs. Cell. 1978 Jul;14(3):695–711. doi: 10.1016/0092-8674(78)90252-0. [DOI] [PubMed] [Google Scholar]
  3. Berk A. J., Sharp P. A. Ultraviolet mapping of the adenovirus 2 early promoters. Cell. 1977 Sep;12(1):45–55. doi: 10.1016/0092-8674(77)90184-2. [DOI] [PubMed] [Google Scholar]
  4. Bonner W. M., Laskey R. A. A film detection method for tritium-labelled proteins and nucleic acids in polyacrylamide gels. Eur J Biochem. 1974 Jul 1;46(1):83–88. doi: 10.1111/j.1432-1033.1974.tb03599.x. [DOI] [PubMed] [Google Scholar]
  5. Chin W. W., Maizel J. V., Jr The polypeptides of adenovirus. VII. Further studies of early polypeptides in vivo and localization of E2 and E2A to the cell plasma membrane. Virology. 1976 Jun;71(2):518–530. doi: 10.1016/0042-6822(76)90378-0. [DOI] [PubMed] [Google Scholar]
  6. Craig E. A., Raskas H. J. Effect of cycloheximide on RNA metabolism early in productive infection with adenovirus 2. J Virol. 1974 Jul;14(1):26–32. doi: 10.1128/jvi.14.1.26-32.1974. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Flint J. The topography and transcription of the adenovirus genome. Cell. 1977 Feb;10(2):153–166. doi: 10.1016/0092-8674(77)90211-2. [DOI] [PubMed] [Google Scholar]
  8. Gallimore P. H. Viral DNA in transformed cells. II. A study of the sequences of adenovirus 2 DNA IN NINE LINES OF TRANSFORMED RAT CELLS USING SPECIFIC FRAGMENTS OF THE VIRAL GENOME;. J Mol Biol. 1974 Oct 15;89(1):49–72. doi: 10.1016/0022-2836(74)90162-4. [DOI] [PubMed] [Google Scholar]
  9. Harter M. L., Lewis J. B. Adenovirus type 2 early proteins synthesized in vitro and in vivo: identification in infected cells of the 38,000- to 50,000- molecular-weight protein encoded by the left end of the adenovirus type 2 genome. J Virol. 1978 Jun;26(3):736–749. doi: 10.1128/jvi.26.3.736-749.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. 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]
  11. Hayman M. J., Crumpton M. J. Isolation of glycoproteins from pig lymphocyte plasma membrane using Lens culinaris phytohemagglutinin. Biochem Biophys Res Commun. 1972 May 26;47(4):923–930. doi: 10.1016/0006-291x(72)90581-5. [DOI] [PubMed] [Google Scholar]
  12. 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]
  13. Ishibashi M., Maizel J. V., Jr The polypeptides of adenovirus. VI. Early and late glycopolypeptides. Virology. 1974 Apr;58(2):345–361. doi: 10.1016/0042-6822(74)90070-1. [DOI] [PubMed] [Google Scholar]
  14. Johansson K., Persson H., Lewis A. M., Pettersson U., Tibbetts C., Philipson L. Viral DNA sequences and gene products in hamster cells transformed by adenovirus type 2. J Virol. 1978 Sep;27(3):628–639. doi: 10.1128/jvi.27.3.628-639.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Kessler S. W. Rapid isolation of antigens from cells with a staphylococcal protein A-antibody adsorbent: parameters of the interaction of antibody-antigen complexes with protein A. J Immunol. 1975 Dec;115(6):1617–1624. [PubMed] [Google Scholar]
  16. Kvist S., Ostberg L., Persson H., Philipson L., Peterson P. A. Molecular association between transplantation antigens and cell surface antigen in adenovirus-transformed cell line. Proc Natl Acad Sci U S A. 1978 Nov;75(11):5674–5678. doi: 10.1073/pnas.75.11.5674. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. LOWRY O. H., ROSEBROUGH N. J., FARR A. L., RANDALL R. J. Protein measurement with the Folin phenol reagent. J Biol Chem. 1951 Nov;193(1):265–275. [PubMed] [Google Scholar]
  18. Lewis A. M., Jr, Rabson A. S., Levine A. S. Studies of nondefective adenovirus 2-simian virus 40 hybrid viruses. Transformation of hamster kidney cells by adenovirus 2 and the nondefective hybrid viruses. J Virol. 1974 Jun;13(6):1291–1301. doi: 10.1128/jvi.13.6.1291-1301.1974. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. 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]
  20. 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]
  21. Neuwald P. D., Meyer J., Maizel J. V., Jr, Westpahl H. Early gene expression of adenovirus type 2: R-loop mapping of mRNA and time course of viral DNA, mRNA, and protein synthesis. J Virol. 1977 Mar;21(3):1019–1030. doi: 10.1128/jvi.21.3.1019-1030.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Parsons J. T., Green M. Biochemical studies on adenovirus multiplication. 18. Resolution of early virus-specific RNA species in Ad 2 infected and transformed cells. Virology. 1971 Jul;45(1):154–162. doi: 10.1016/0042-6822(71)90122-x. [DOI] [PubMed] [Google Scholar]
  23. Persson H., Oberg B., Philipson L. Purification and characterization of an early protein (E14K) from adenovirus type 2-infected cells. J Virol. 1978 Oct;28(1):119–139. doi: 10.1128/jvi.28.1.119-139.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Persson H., Pettersson U., Mathews M. B. Synthesis of a structural adenovirus polypeptide in the absence of viral DNA replication. Virology. 1978 Oct 1;90(1):67–79. doi: 10.1016/0042-6822(78)90334-3. [DOI] [PubMed] [Google Scholar]
  25. 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]
  26. Saborio J. L., Oberg B. In vivo and in vitro synthesis of adenovirus type 2 early proteins. J Virol. 1976 Mar;17(3):865–875. doi: 10.1128/jvi.17.3.865-875.1976. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Sharon N., Lis H. Lectins: cell-agglutinating and sugar-specific proteins. Science. 1972 Sep 15;177(4053):949–959. doi: 10.1126/science.177.4053.949. [DOI] [PubMed] [Google Scholar]
  28. 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]
  29. Walter G., Maizel J. V., Jr The polypeptides of adenovirus. IV. Detection of early and late virus-induced polypeptides and their distribution in subcellular fractions. Virology. 1974 Feb;57(2):402–408. doi: 10.1016/0042-6822(74)90180-9. [DOI] [PubMed] [Google Scholar]
  30. Zinkernagel R. M., Doherty P. C. The concept that surveillance of self is mediated via the same set of genes that determines recognition of allogenic cells. Cold Spring Harb Symp Quant Biol. 1977;41(Pt 2):505–510. doi: 10.1101/sqb.1977.041.01.058. [DOI] [PubMed] [Google Scholar]

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