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. 1982 Jul;43(1):73–82. doi: 10.1128/jvi.43.1.73-82.1982

Discrete regions of simian virus 40 large T antigen are required for nonspecific and viral origin-specific DNA binding.

C Prives, B Barnet, A Scheller, G Khoury, G Jay
PMCID: PMC256098  PMID: 6287017

Abstract

The nondefective adenovirus type 2 (Ad2)-simian virus 40 (SV40) hybrid viruses, Ad2+ND2 and Ad2+ND4, have been used to determine which regions of the SV40 genome coding for the large tumor (T) antigen are involved in specific and nonspecific DNA binding. Ad2+ND2 encodes 45,000 M4 (45K) and 56,000 Mr (56K) T antigen-related polypeptides. The 45K polypeptide did not bind to DNA, but the 56K polypeptide bound nonspecifically to calf thymus DNA, Ad2+ND4 encodes 50,000 Mr (60K), 66,000 Mr (66K), 70,000 Mr (70K), 74,000 Mr (74K), and 90,000 Mr (90K) T antigen-related polypeptides, all of which bound nonspecifically to calf thymus DNA. However, in more stringent assays, where tight binding to viral origin sequences was tested, only the 90K protein specified by Ad2A+ND4 showed specific high affinity for sequences at the viral origin of replication. From these results and previously published experiments describing the SV40 DNA integrated into these hybrid viruses, it was concluded that SV40 early gene sequences located between 0.39 and 0.44 SV40 map units contribute to nonspecific DNA binding, whereas sequences located between 0.50 and 0.63 SV40 map units are necessary for specific binding to the viral origin of replication.

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Selected References

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

  1. Berk A. J., Sharp P. A. Spliced early mRNAs of simian virus 40. Proc Natl Acad Sci U S A. 1978 Mar;75(3):1274–1278. doi: 10.1073/pnas.75.3.1274. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Carroll R. B., Hager L., Dulbecco R. Simian virus 40 T antigen binds to DNA. Proc Natl Acad Sci U S A. 1974 Sep;71(9):3754–3757. doi: 10.1073/pnas.71.9.3754. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Deppert W., Walter G. Simian virus to (SV40) tumor-specific proteins in nucleus and plasma membrane of HeLa cells infected by adenovirus 2-SV40 hybrid virus Ad2+ND2. Proc Natl Acad Sci U S A. 1976 Jul;73(7):2505–2509. doi: 10.1073/pnas.73.7.2505. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Fiers W., Contreras R., Haegemann G., Rogiers R., Van de Voorde A., Van Heuverswyn H., Van Herreweghe J., Volckaert G., Ysebaert M. Complete nucleotide sequence of SV40 DNA. Nature. 1978 May 11;273(5658):113–120. doi: 10.1038/273113a0. [DOI] [PubMed] [Google Scholar]
  5. Gluzman Y., Davison J., Oren M., Winocour E. Properties of permissive monkey cells transformed by UV-irradiated simian virus 40. J Virol. 1977 May;22(2):256–266. doi: 10.1128/jvi.22.2.256-266.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Gurney E. G., Harrison R. O., Fenno J. Monoclonal antibodies against simian virus 40 T antigens: evidence for distinct sublcasses of large T antigen and for similarities among nonviral T antigens. J Virol. 1980 Jun;34(3):752–763. doi: 10.1128/jvi.34.3.752-763.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Jay G., Ferrini U., Robinson E. A., Khoury G., Appella E. Cell-free synthesis of mouse H-2 histocompatibility antigens. Proc Natl Acad Sci U S A. 1979 Dec;76(12):6562–6566. doi: 10.1073/pnas.76.12.6562. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Jay G., Jay F. T., Friedman R. M., Levine A. S. Biosynthesis, immunological specificity, and intracellular distribution of the simian virus 40-specific protein induced by the nondefective hybrid Ad2+ND1. J Virol. 1978 May;26(2):411–419. doi: 10.1128/jvi.26.2.411-419.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Jessel D., Landau T., Hudson J., Lalor T., Tenen D., Livingston D. M. Identification of regions of the SV40 genome which contain preferred SV40 T antigen-binding sites. Cell. 1976 Aug;8(4):535–545. doi: 10.1016/0092-8674(76)90222-1. [DOI] [PubMed] [Google Scholar]
  10. Khoury G., Alwine J., Goldman N., Gruss P., Jay G. New chimeric splice junction in adenovirus type 2-simian virus 40 hybrid viral mRNA. J Virol. 1980 Oct;36(1):143–151. doi: 10.1128/jvi.36.1.143-151.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Lane D. P., Crawford L. V. T antigen is bound to a host protein in SV40-transformed cells. Nature. 1979 Mar 15;278(5701):261–263. doi: 10.1038/278261a0. [DOI] [PubMed] [Google Scholar]
  12. Lawrence C., Hunter T., Walter G. Characterization of the simian virus 40-specific messenger RNAs isolated from HeLa cells infected with the non-defective adenovirus 2-simian virus 40 hybrid viruses Ad2+ND2 and Ad2+ND4. J Mol Biol. 1979 Oct 15;134(1):95–108. doi: 10.1016/0022-2836(79)90415-7. [DOI] [PubMed] [Google Scholar]
  13. Levine A. J., Ginsberg H. S. Role of adenovirus structural proteins in the cessation of host-cell biosynthetic functions. J Virol. 1968 May;2(5):430–439. doi: 10.1128/jvi.2.5.430-439.1968. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Lewis A. M., Jr Defective and nondefective Ad2-SV40 hybrids. Prog Med Virol. 1977;23:96–139. [PubMed] [Google Scholar]
  15. Linke H. K., Hunter T., Walter G. Structural relationship between the 100,000- and 17,000- molecular-weight T antigens of simian virus 40 (SV40) as deduced by comparison with the SV40-specific proteins coded by the nondefective adenovirus type 2-SV40 hybrid viruses. J Virol. 1979 Jan;29(1):390–394. doi: 10.1128/jvi.29.1.390-394.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Livingston D. M., Henderson I. C., Hudson J. SV40 T antigen: partial purification and properties. Cold Spring Harb Symp Quant Biol. 1975;39(Pt 1):283–289. doi: 10.1101/sqb.1974.039.01.037. [DOI] [PubMed] [Google Scholar]
  17. Mann K., Hunter T., Walter G., Linke H. Evidence for simian virus 40 (SV40) coding of SV40 T-antigen and the SV40-specific proteins in HeLa cells infected with nondefective adenovirus type 2-SV40 hybrid viruses. J Virol. 1977 Oct;24(1):151–169. doi: 10.1128/jvi.24.1.151-169.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. McKay R. D. Binding of a simian virus 40 T antigen-related protein to DNA. J Mol Biol. 1981 Jan 25;145(3):471–488. doi: 10.1016/0022-2836(81)90540-4. [DOI] [PubMed] [Google Scholar]
  19. Melero J. A., Stitt D. T., Mangel W. F., Carroll R. B. Identification of new polypeptide species (48-55K) immunoprecipitable by antiserum to purified large T antigen and present in SV40-infected and -transformed cells. Virology. 1979 Mar;93(2):466–480. doi: 10.1016/0042-6822(79)90250-2. [DOI] [PubMed] [Google Scholar]
  20. Morrow J. F., Berg P., Kelly T. J., Jr, Lewis A. M., Jr Mapping of simian virus 40 early functions on the viral chromosome. J Virol. 1973 Sep;12(3):653–658. doi: 10.1128/jvi.12.3.653-658.1973. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Myers R. M., Williams R. C., Tjian R. Oligomeric structure of a simian virus 40 T antigen in free form and bound to DNA. J Mol Biol. 1981 Jun 5;148(4):347–353. doi: 10.1016/0022-2836(81)90180-7. [DOI] [PubMed] [Google Scholar]
  22. Oren M., Winocour E., Prives C. Differential affinities of simian virus 40 large tumor antigen for DNA. Proc Natl Acad Sci U S A. 1980 Jan;77(1):220–224. doi: 10.1073/pnas.77.1.220. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Osborn M., Weber K. SV40: T antigen, the A function and transformation. Cold Spring Harb Symp Quant Biol. 1975;39(Pt 1):267–276. doi: 10.1101/sqb.1974.039.01.035. [DOI] [PubMed] [Google Scholar]
  24. Oxman M. N., Levin M. J., Lewis A. M., Jr Control of simian virus 40 gene expression in adenovirus-simian virus 40 hybrid viruses. Synthesis of hybrid adenovirus 2-simian virus 40 RNA molecules in cells infected with a nondefective adenovirus 2-simian virus 40 hybrid virus. J Virol. 1974 Feb;13(2):322–330. doi: 10.1128/jvi.13.2.322-330.1974. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Prives C., Beck Y., Shure H. DNA binding properties of simian virus 40 T-antigens synthesized in vivo and in vitro. J Virol. 1980 Feb;33(2):689–696. doi: 10.1128/jvi.33.2.689-696.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Reddy V. B., Thimmappaya B., Dhar R., Subramanian K. N., Zain B. S., Pan J., Ghosh P. K., Celma M. L., Weissman S. M. The genome of simian virus 40. Science. 1978 May 5;200(4341):494–502. doi: 10.1126/science.205947. [DOI] [PubMed] [Google Scholar]
  27. Reed S. I., Ferguson J., Davis R. W., Stark G. R. T antigen binds to simian virus 40 DNA at the origin of replication. Proc Natl Acad Sci U S A. 1975 Apr;72(4):1605–1609. doi: 10.1073/pnas.72.4.1605. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Reed S. I., Stark G. R., Alwine J. C. Autoregulation of simian virus 40 gene A by T antigen. Proc Natl Acad Sci U S A. 1976 Sep;73(9):3083–3087. doi: 10.1073/pnas.73.9.3083. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Rigby P. W., Dieckmann M., Rhodes C., Berg P. Labeling deoxyribonucleic acid to high specific activity in vitro by nick translation with DNA polymerase I. J Mol Biol. 1977 Jun 15;113(1):237–251. doi: 10.1016/0022-2836(77)90052-3. [DOI] [PubMed] [Google Scholar]
  30. Rundell K., Collins J. K., Tegtmeyer P., Ozer H. L., Lai C. J., Nathans D. Identification of simian virus 40 protein A. J Virol. 1977 Feb;21(2):636–646. doi: 10.1128/jvi.21.2.636-646.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Shalloway D., Kleinberger T., Livingston D. M. Mapping of SV40 DNA replication origin region binding sites for the SV40 T antigen by protection against exonuclease III digestion. Cell. 1980 Jun;20(2):411–422. doi: 10.1016/0092-8674(80)90627-3. [DOI] [PubMed] [Google Scholar]
  32. Shortle D. R., Margolskee R. F., Nathans D. Mutational analysis of the simian virus 40 replicon: pseudorevertants of mutants with a defective replication origin. Proc Natl Acad Sci U S A. 1979 Dec;76(12):6128–6131. doi: 10.1073/pnas.76.12.6128. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Tegtmeyer P. Simian virus 40 deoxyribonucleic acid synthesis: the viral replicon. J Virol. 1972 Oct;10(4):591–598. doi: 10.1128/jvi.10.4.591-598.1972. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Thummel C., Tjian R., Grodzicker T. Expression of SV40 T antigen under control of adenovirus promoters. Cell. 1981 Mar;23(3):825–836. doi: 10.1016/0092-8674(81)90447-5. [DOI] [PubMed] [Google Scholar]
  35. Tjian R., Robbins A. Enzymatic activities associated with a purified simian virus 40 T antigen-related protein. Proc Natl Acad Sci U S A. 1979 Feb;76(2):610–614. doi: 10.1073/pnas.76.2.610. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Westphal H. Genomic arrangement of an adenovirus-simian virus 40 hybrid virus, Ad2+ND4del. J Virol. 1981 Nov;40(2):526–532. doi: 10.1128/jvi.40.2.526-532.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. Westphal H., Lai S. P., Lawrence C., Hunter T., Walter G. Mosaic adenovirus-SV40 RNA specified by the non-defective hybrid virus Ad2+ND4. J Mol Biol. 1979 May 25;130(3):337–351. doi: 10.1016/0022-2836(79)90545-x. [DOI] [PubMed] [Google Scholar]

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