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. 1989 May;63(5):2308–2316. doi: 10.1128/jvi.63.5.2308-2316.1989

Nuclear subcompartmentalization of simian virus 40 large T antigen: evidence for in vivo regulation of biochemical activities.

R Schirmbeck 1, W Deppert 1
PMCID: PMC250649  PMID: 2539520

Abstract

Simian virus 40 large T antigen (large T) in the early and the late phases of infection differs significantly in its sequence-specific DNA-binding and ATPase activities, indicating that different large-T populations participate in virus-specific events at various stages of the infectious cycle. To further characterize these large-T populations, we have analyzed nuclear subclasses of large T, isolated from their in vivo location, for their biochemical activities. We show that chromatin- and nuclear matrix-associated large-T molecules exhibit different simian virus 40 control region (ORI) DNA-binding and ATPase activities. The association of large T with a certain nuclear substructure, therefore, subcompartmentalizes large-T molecules exerting different biochemical activities. Nuclear subcompartmentalization thus may provide a higher-order level for the regulation of biochemical activities of large T in vivo.

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Selected References

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  1. Buckler-White A. J., Humphrey G. W., Pigiet V. Association of polyoma T antigen and DNA with the nuclear matrix from lytically infected 3T6 cells. Cell. 1980 Nov;22(1 Pt 1):37–46. doi: 10.1016/0092-8674(80)90152-x. [DOI] [PubMed] [Google Scholar]
  2. Butel J. S., Jarvis D. L. The plasma-membrane-associated form of SV40 large tumor antigen: biochemical and biological properties. Biochim Biophys Acta. 1986 Oct 28;865(2):171–195. doi: 10.1016/0304-419x(86)90027-2. [DOI] [PubMed] [Google Scholar]
  3. Caron de Fromentel C., Viron A., Puvion E., May P. SV40 large T-antigen and transformation related protein p53 are associated in situ with nuclear RNP structures containing hnRNA of transformed cells. Exp Cell Res. 1986 May;164(1):35–48. doi: 10.1016/0014-4827(86)90452-0. [DOI] [PubMed] [Google Scholar]
  4. Clark R., Lane D. P., Tjian R. Use of monoclonal antibodies as probes of simian virus 40 T antigen ATPase activity. J Biol Chem. 1981 Nov 25;256(22):11854–11858. [PubMed] [Google Scholar]
  5. DiMaio D., Nathans D. Regulatory mutants of simian virus 40. Effect of mutations at a T antigen binding site on DNA replication and expression of viral genes. J Mol Biol. 1982 Apr 15;156(3):531–548. doi: 10.1016/0022-2836(82)90265-0. [DOI] [PubMed] [Google Scholar]
  6. Fanning E., Westphal K. H., Brauer D., Cörlin D. Subclasses of simian virus 40 large T antigen: differential binding of two subclasses of T antigen from productively infected cells to viral and cellular DNA. EMBO J. 1982;1(9):1023–1028. doi: 10.1002/j.1460-2075.1982.tb01290.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Gannon J. V., Lane D. P. p53 and DNA polymerase alpha compete for binding to SV40 T antigen. Nature. 1987 Oct 1;329(6138):456–458. doi: 10.1038/329456a0. [DOI] [PubMed] [Google Scholar]
  8. Giacherio D., Hager L. P. A poly(dT)-stimulated ATPase activity associated with simian virus 40 large T antigen. J Biol Chem. 1979 Sep 10;254(17):8113–8116. [PubMed] [Google Scholar]
  9. Grässer F. A., Mann K., Walter G. Removal of serine phosphates from simian virus 40 large T antigen increases its ability to stimulate DNA replication in vitro but has no effect on ATPase and DNA binding. J Virol. 1987 Nov;61(11):3373–3380. doi: 10.1128/jvi.61.11.3373-3380.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Gurney E. G., Tamowski S., Deppert W. Antigenic binding sites of monoclonal antibodies specific for simian virus 40 large T antigen. J Virol. 1986 Mar;57(3):1168–1172. doi: 10.1128/jvi.57.3.1168-1172.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Harper F., Florentin Y., Puvion E. Large T antigen-rich viral DNA replication loci in SV40-infected monkey kidney cells. Exp Cell Res. 1985 Dec;161(2):434–444. doi: 10.1016/0014-4827(85)90099-0. [DOI] [PubMed] [Google Scholar]
  12. Hinzpeter M., Deppert W. Analysis of biological and biochemical parameters for chromatin and nuclear matrix association of SV40 large T antigen in transformed cells. Oncogene. 1987 May;1(2):119–129. [PubMed] [Google Scholar]
  13. Hinzpeter M., Fanning E., Deppert W. A new sensitive target-bound DNA binding assay for SV40 large T antigen. Virology. 1986 Jan 15;148(1):159–167. doi: 10.1016/0042-6822(86)90411-3. [DOI] [PubMed] [Google Scholar]
  14. Hirt B. Selective extraction of polyoma DNA from infected mouse cell cultures. J Mol Biol. 1967 Jun 14;26(2):365–369. doi: 10.1016/0022-2836(67)90307-5. [DOI] [PubMed] [Google Scholar]
  15. 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]
  16. MacArthur H., Walter G. Monoclonal antibodies specific for the carboxy terminus of simian virus 40 large T antigen. J Virol. 1984 Nov;52(2):483–491. doi: 10.1128/jvi.52.2.483-491.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Manos M. M., Gluzman Y. Genetic and biochemical analysis of transformation-competent, replication-defective simian virus 40 large T antigen mutants. J Virol. 1985 Jan;53(1):120–127. doi: 10.1128/jvi.53.1.120-127.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Margolskee R. F., Nathans D. Simian virus 40 mutant T antigens with relaxed specificity for the nucleotide sequence at the viral DNA origin of replication. J Virol. 1984 Feb;49(2):386–393. doi: 10.1128/jvi.49.2.386-393.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Martin R. G. The transformation of cell growth and transmogrification of DNA synthesis by simian virus 40. Adv Cancer Res. 1981;34:1–68. doi: 10.1016/s0065-230x(08)60238-9. [DOI] [PubMed] [Google Scholar]
  20. McCormick F., Harlow E. Association of a murine 53,000-dalton phosphoprotein with simian virus 40 large-T antigen in transformed cells. J Virol. 1980 Apr;34(1):213–224. doi: 10.1128/jvi.34.1.213-224.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Mitchell P. J., Wang C., Tjian R. Positive and negative regulation of transcription in vitro: enhancer-binding protein AP-2 is inhibited by SV40 T antigen. Cell. 1987 Sep 11;50(6):847–861. doi: 10.1016/0092-8674(87)90512-5. [DOI] [PubMed] [Google Scholar]
  22. Mohr I. J., Stillman B., Gluzman Y. Regulation of SV40 DNA replication by phosphorylation of T antigen. EMBO J. 1987 Jan;6(1):153–160. doi: 10.1002/j.1460-2075.1987.tb04733.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Mole S. E., Gannon J. V., Ford M. J., Lane D. P. Structure and function of SV40 large-T antigen. Philos Trans R Soc Lond B Biol Sci. 1987 Dec 15;317(1187):455–469. doi: 10.1098/rstb.1987.0072. [DOI] [PubMed] [Google Scholar]
  24. Myers R. M., Rio D. C., Robbins A. K., Tjian R. SV40 gene expression is modulated by the cooperative binding of T antigen to DNA. Cell. 1981 Aug;25(2):373–384. doi: 10.1016/0092-8674(81)90056-8. [DOI] [PubMed] [Google Scholar]
  25. Rio D., Robbins A., Myers R., Tjian R. Regulation of simian virus 40 early transcription in vitro by a purified tumor antigen. Proc Natl Acad Sci U S A. 1980 Oct;77(10):5706–5710. doi: 10.1073/pnas.77.10.5706. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Robb J. A., Huebner K. Effect of cell chromosome number on simian virus 40 replication. Exp Cell Res. 1973 Sep;81(1):120–126. doi: 10.1016/0014-4827(73)90118-3. [DOI] [PubMed] [Google Scholar]
  27. Runzler R., Thompson S., Fanning E. Oligomerization and origin DNA-binding activity of simian virus 40 large T antigen. J Virol. 1987 Jul;61(7):2076–2083. doi: 10.1128/jvi.61.7.2076-2083.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Ryder K., Silver S., DeLucia A. L., Fanning E., Tegtmeyer P. An altered DNA conformation in origin region I is a determinant for the binding of SV40 large T antigen. Cell. 1986 Mar 14;44(5):719–725. doi: 10.1016/0092-8674(86)90838-x. [DOI] [PubMed] [Google Scholar]
  29. Ryder K., Vakalopoulou E., Mertz R., Mastrangelo I., Hough P., Tegtmeyer P., Fanning E. Seventeen base pairs of region I encode a novel tripartite binding signal for SV40 T antigen. Cell. 1985 Sep;42(2):539–548. doi: 10.1016/0092-8674(85)90111-4. [DOI] [PubMed] [Google Scholar]
  30. Scheidtmann K. H., Hardung M., Echle B., Walter G. DNA-binding activity of simian virus 40 large T antigen correlates with a distinct phosphorylation state. J Virol. 1984 Apr;50(1):1–12. doi: 10.1128/jvi.50.1.1-12.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Schirmbeck R., Deppert W. Analysis of mechanisms controlling the interactions of SV40 large T antigen with the SV40 ORI region. Virology. 1988 Aug;165(2):527–538. doi: 10.1016/0042-6822(88)90597-1. [DOI] [PubMed] [Google Scholar]
  32. Schirmbeck R., Deppert W. Specific interaction of simian virus 40 large T antigen with cellular chromatin and nuclear matrix during the course of infection. J Virol. 1987 Nov;61(11):3561–3569. doi: 10.1128/jvi.61.11.3561-3569.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. 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]
  34. Simanis V., Lane D. P. An immunoaffinity purification procedure for SV40 large T antigen. Virology. 1985 Jul 15;144(1):88–100. doi: 10.1016/0042-6822(85)90308-3. [DOI] [PubMed] [Google Scholar]
  35. Smale S. T., Tjian R. T-antigen-DNA polymerase alpha complex implicated in simian virus 40 DNA replication. Mol Cell Biol. 1986 Nov;6(11):4077–4087. doi: 10.1128/mcb.6.11.4077. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Smith H. C., Berezney R., Brewster J. M., Rekosh D. Properties of adenoviral DNA bound to the nuclear matrix. Biochemistry. 1985 Feb 26;24(5):1197–1202. doi: 10.1021/bi00326a022. [DOI] [PubMed] [Google Scholar]
  37. Stahl H., Knippers R. The simian virus 40 large tumor antigen. Biochim Biophys Acta. 1987 Oct 9;910(1):1–10. doi: 10.1016/0167-4781(87)90088-1. [DOI] [PubMed] [Google Scholar]
  38. Staufenbiel M., Deppert W. Different structural systems of the nucleus are targets for SV40 large T antigen. Cell. 1983 May;33(1):173–181. doi: 10.1016/0092-8674(83)90346-x. [DOI] [PubMed] [Google Scholar]
  39. Staufenbiel M., Deppert W. Preparation of nuclear matrices from cultured cells: subfractionation of nuclei in situ. J Cell Biol. 1984 May;98(5):1886–1894. doi: 10.1083/jcb.98.5.1886. [DOI] [PMC free article] [PubMed] [Google Scholar]
  40. Tack L. C., Wright J. H., Gurney E. G. Characterization of simian virus 40 large T antigen by using different monoclonal antibodies: T-p53 complexes are preferentially ATPase active and adenylylated. J Virol. 1988 Mar;62(3):1028–1037. doi: 10.1128/jvi.62.3.1028-1037.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  41. 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]
  42. Verderame M. F., Kohtz D. S., Pollack R. E. 94,000- and 100,000-molecular-weight simian virus 40 T-antigens are associated with the nuclear matrix in transformed and revertant mouse cells. J Virol. 1983 May;46(2):575–583. doi: 10.1128/jvi.46.2.575-583.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  43. Voelkerding K., Klessig D. F. Identification of two nuclear subclasses of the adenovirus type 5-encoded DNA-binding protein. J Virol. 1986 Nov;60(2):353–362. doi: 10.1128/jvi.60.2.353-362.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  44. Wilson V. G., Tevethia M. J., Lewton B. A., Tegtmeyer P. DNA binding properties of simian virus 40 temperature-sensitive A proteins. J Virol. 1982 Nov;44(2):458–466. doi: 10.1128/jvi.44.2.458-466.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  45. van der Velden H. M., Wanka F. The nuclear matrix--its role in the spatial organization and replication of eukaryotic DNA. Mol Biol Rep. 1987;12(2):69–77. doi: 10.1007/BF00368873. [DOI] [PubMed] [Google Scholar]

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