Skip to main content
PMC home page
Journal of Virology logoLink to Journal of Virology
. 1978 Jun;26(3):736–749. doi: 10.1128/jvi.26.3.736-749.1978

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.

M L Harter, J B Lewis
PMCID: PMC525899  PMID: 671586

Abstract

Analysis by sodium dodecyl sulfate-polyacrylamide gel electrophoresis of protein synthesized early after injection of human cells with adenovirus type 2 (Ad2) showed that polypeptides of 11,000 (11K), 12K, 14K, 15K, 19K, 21K, 24K, and 72k molecular weight were present in infected but not in mock-infected cells. These polypeptides corresponded in electrophoretic mobility to the following polypeptides synthesized in vitro by using mRNA complementary to specific regions of the Ad2 genome: 11K, 19K, 21K (91.5 to 96.8 map units), 14K (78.2 to 83.4 map units), 72K (62.4 to 67.9 map units), and 15K (4.9 to 11.0 map units). Polypeptides of 25K, 17K, 15.5K, and 13K were also synthesized in vitro, but have not yet been detected in infected cells. In addition, six adeno-specific polypeptides of 38 to 50K molecular weight could be discerned in infected cells if two-dimensional isoelectric focusing-polyacrylamide gel electrophoresis was used to compare extracts from infected and mock-infected cells. Partial protease digestion showed these 38 to 50K polypeptides to be related in sequence to each other and to the 40 to 50K polypeptide made in vitro (1.3 to 4.0 map units).

Full text

PDF
736

Images in this article

739Image
on p.739
741Image
on p.741
742Image
on p.742
744Image
on p.744
746Image
on p.746

Selected References

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

  1. Anderson C. W., Lewis J. B., Atkins J. F., Gesteland R. F. Cell-free synthesis of adenovirus 2 proteins programmed by fractionated messenger RNA: a comparison of polypeptide products and messenger RNA lengths. Proc Natl Acad Sci U S A. 1974 Jul;71(7):2756–2760. doi: 10.1073/pnas.71.7.2756. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Atkins J. F., Lewis J. B., Anderson C. W., Gesteland R. F. Enhanced differential synthesis of proteins in a mammalian cell-free system by addition of polyamines. J Biol Chem. 1975 Jul 25;250(14):5688–5695. [PubMed] [Google Scholar]
  3. 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]
  4. Chow L. T., Roberts J. M., Lewis J. B., Broker T. R. A map of cytoplasmic RNA transcripts from lytic adenovirus type 2, determined by electron microscopy of RNA:DNA hybrids. Cell. 1977 Aug;11(4):819–836. doi: 10.1016/0092-8674(77)90294-x. [DOI] [PubMed] [Google Scholar]
  5. Cleveland D. W., Fischer S. G., Kirschner M. W., Laemmli U. K. Peptide mapping by limited proteolysis in sodium dodecyl sulfate and analysis by gel electrophoresis. J Biol Chem. 1977 Feb 10;252(3):1102–1106. [PubMed] [Google Scholar]
  6. Dunn A. R., Hassell J. A. A novel method to map transcripts: evidence for homology between an adenovirus mRNA and discrete multiple regions of the viral genome. Cell. 1977 Sep;12(1):23–36. doi: 10.1016/0092-8674(77)90182-9. [DOI] [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. Flint S. J., Gallimore P. H., Sharp P. A. Comparison of viral RNA sequences in adenovirus 2-transformed and lytically infected cells. J Mol Biol. 1975 Jul 25;96(1):47–68. doi: 10.1016/0022-2836(75)90181-3. [DOI] [PubMed] [Google Scholar]
  9. Gilead Z., Jeng Y. H., Wold W. S., Sugawara K., Rho H. M., Harter M. L., Green M. Immunological identification of two adenovirus 2-induced early proteins possibly involved in cell transformation. Nature. 1976 Nov 18;264(5583):263–266. doi: 10.1038/264263a0. [DOI] [PubMed] [Google Scholar]
  10. Graham F. L., Abrahams P. J., Mulder C., Heijneker H. L., Warnaar S. O., De Vries F. A., Fiers W., Van Der Eb A. J. Studies on in vitro transformation by DNA and DNA fragments of human adenoviruses and simian virus 40. Cold Spring Harb Symp Quant Biol. 1975;39(Pt 1):637–650. doi: 10.1101/sqb.1974.039.01.077. [DOI] [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. Klessig D. F. Two adenovirus mRNAs have a common 5' terminal leader sequence encoded at least 10 kb upstream from their main coding regions. Cell. 1977 Sep;12(1):9–21. doi: 10.1016/0092-8674(77)90181-7. [DOI] [PubMed] [Google Scholar]
  13. 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]
  14. Levinson A., Levine A. J. The isolation and identification of the adenovirus group C tumor antigens. Virology. 1977 Jan;76(1):1–11. doi: 10.1016/0042-6822(77)90275-6. [DOI] [PubMed] [Google Scholar]
  15. Lewis J. B., Anderson C. W., Atkins J. F. Further mapping of late adenovirus genes by cell-free translation of RNA selected by hybridization to specific DNA fragments. Cell. 1977 Sep;12(1):37–44. doi: 10.1016/0092-8674(77)90183-0. [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. Oberg B., Saborio J., Persson T., Everitt E., Philipson L. Identification of the in vitro translation products of adenovirus mRNA by immunoprecipitation. J Virol. 1975 Jan;15(1):199–207. doi: 10.1128/jvi.15.1.199-207.1975. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. 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]
  19. Schick J., Baczko K., Fanning E., Groneberg J., Burger H., Doerfler W. Intracellular forms of adenovirus DNA: integrated form of adenovirus DNA appears early in productive infection. Proc Natl Acad Sci U S A. 1976 Apr;73(4):1043–1047. doi: 10.1073/pnas.73.4.1043. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Weiner A. M., Weber K. A single UGA codon functions as a natural termination signal in the coliphage q beta coat protein cistron. J Mol Biol. 1973 Nov 15;80(4):837–855. doi: 10.1016/0022-2836(73)90213-1. [DOI] [PubMed] [Google Scholar]

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

RESOURCES