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. 1981 Jan;37(1):524–529. doi: 10.1128/jvi.37.1.524-529.1981

In vitro synthesis of adenovirus type 5 T antigens. I. Translation of early region 1-specific rna from lytically infected cells.

J H Lupker, A Davis, H Jochemsen, A J van der Eb
PMCID: PMC171033  PMID: 7218430

Abstract

Translation of early RNA specific for the leftmost early region 1 of adenovirus type 5 DNA in a rabbit reticulocyte cell-free system resulted in the synthesis of proteins with the following molecular weights: 65,000 (65K), 54K, 42 to 34K, 29K, 25K, 19K, and 18K. All of these proteins could be immunoprecipitated with hamster antitumor serum. The 42 to 34K proteins mapped in early region 1a, and those with molecular weights of 65K, 19K, and 18K mapped in early region 1b. The gene coding for the 54K protein may be localized outside of early region 1. We could not map unambiguously the 29K and 25K proteins. The identification of a 65K protein among products synthesized in vitro suggests that this protein may be identical to the 65K major T antigen present in adenovirus type 5-infected and -transformed cells, and this indicates that it is indeed encoded by the viral genome. This protein is encoded by a 23S mRNA. The other early region 1-specific proteins appear to be encoded by mRNA of approximately 13S, except for the 19K protein, which is synthesized with RNA sedimenting at both 13S and 23S.

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Selected References

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  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. 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. Büttner W., Veres-Molnár Z., Green M. Isolation of DNA Strand-specific early messenger RNA species in cells infected by human adenovirus 2. Proc Natl Acad Sci U S A. 1974 Aug;71(8):2951–2955. doi: 10.1073/pnas.71.8.2951. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Chow L. T., Broker T. R., Lewis J. B. Complex splicing patterns of RNAs from the early regions of adenovirus-2. J Mol Biol. 1979 Oct 25;134(2):265–303. doi: 10.1016/0022-2836(79)90036-6. [DOI] [PubMed] [Google Scholar]
  5. Davies J. W., Kaesberg P. Translation of virus mRNA: protein synthesis directed by several virus RNAs in a cell-free extract from wheat germ. J Gen Virol. 1974 Oct;25(1):11–20. doi: 10.1099/0022-1317-25-1-11. [DOI] [PubMed] [Google Scholar]
  6. Flint S. J., Sambrook J., Williams J. F., Sharp P. A. Viral nucleic acid sequences in transformed cells. IV. A study of the sequences of adenovirus 5 DNA and RNA in four lines of adenovirus 5-transformed rodent cells using specific fragments of the viral genome. Virology. 1976 Jul 15;72(2):456–470. doi: 10.1016/0042-6822(76)90174-4. [DOI] [PubMed] [Google Scholar]
  7. 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]
  8. Gillespie D., Spiegelman S. A quantitative assay for DNA-RNA hybrids with DNA immobilized on a membrane. J Mol Biol. 1965 Jul;12(3):829–842. doi: 10.1016/s0022-2836(65)80331-x. [DOI] [PubMed] [Google Scholar]
  9. 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]
  10. Halbert D. N., Spector D. J., Raskas H. J. In vitro translation products specified by the transforming region of adenovirus type 2. J Virol. 1979 Sep;31(3):621–629. doi: 10.1128/jvi.31.3.621-629.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Levinson A. D., Levine A. J. The group C adenovirus tumor antigens: identification in infected and transformed cells and a peptide map analysis. Cell. 1977 Aug;11(4):871–879. doi: 10.1016/0092-8674(77)90298-7. [DOI] [PubMed] [Google Scholar]
  12. 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]
  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. Maat J., van Beveren C. P., van Ormondt H. The nucleotide sequence of adenovirus type 5 early region E1: the region between map positions 8.0 (HindIII site) and 11.8 (SmaI site). Gene. 1980 Jun;10(1):27–38. doi: 10.1016/0378-1119(80)90140-7. [DOI] [PubMed] [Google Scholar]
  15. Noyes B. E., Stark G. R. Nucleic acid hybridization using DNA covalently coupled to cellulose. Cell. 1975 Jul;5(3):301–310. doi: 10.1016/0092-8674(75)90105-1. [DOI] [PubMed] [Google Scholar]
  16. Persson H., Mathisen B., Philipson L., Pettersson U. A maturation protein in adenovirus morphogenesis. Virology. 1979 Feb;93(1):198–208. doi: 10.1016/0042-6822(79)90287-3. [DOI] [PubMed] [Google Scholar]
  17. Schrier P. I., Van Den Elsen P. J., Hertoghs J. J., Van Der Eb A. J. Characterization of tumor antigens in cells transformed by fragments of adenovirus type 5 DNA. Virology. 1979 Dec;99(2):372–385. doi: 10.1016/0042-6822(79)90016-3. [DOI] [PubMed] [Google Scholar]
  18. Van der Eb A. J., Mulder C., Graham F. L., Houweling A. Transformation with specific fragments of adenovirus DNAs. I. Isolation of specific fragments with transforming activity of adenovirus 2 and 5 DNA. Gene. 1977;2(3-4):115–132. doi: 10.1016/0378-1119(77)90012-9. [DOI] [PubMed] [Google Scholar]

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