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The Quaternary Structures of SV40 Large T Antigen and Tumor Suppressor p53: Analysis by Gel Electrophoresis

  • Judith E. Stenger
  • Gregory A. Mayr
  • Kristine Mann
  • Satyajit Ray
  • Mary E. Anderson
  • Peter Tegtmeyer
Part of the Genetic Engineering book series (GEPM, volume 14)

Abstract

Protein-protein interactions have become a major theme in molecular biology. In many cases, the homo-oligomeric structure of a protein is essential for the activity of that protein. For example, both SV40 large T antigen and the cellular tumor suppressor p53 have distinctive quaternary structures that determine their intrinsic functions. In other cases, hetero-oligomeric structures lead to the gain or loss of function of the interacting proteins. Again, T antigen and p53 are excellent examples of this kind of interaction. T antigen-binding blocks the function of p53 in the regulation of cellular proliferation (1–3), and p53 in turn may modulate T antigen’s replication functions (4–6). In view of the obvious importance of protein-protein interactions, simple techniques for the analysis of protein oligomers will be crucial to an understanding of their functions. Here, we review the use of gradient gel electrophoresis and protein crosslinking to analyze the quaternary structures of SV40 large T antigen, murine p53, and their complexes. This technique offers several advantages over conventional gradient centrifugation for analyzing quaternary structure. Multiple samples can be examined in parallel with resolution superior to that of gradient centrifugation. We have used purified proteins produced by a baculovirus expression system. The methods for these studies have been described previously (7,8).

Keywords

Zinc Finger Quaternary Structure Oligomeric Form Oligomeric Structure Hexamer Formation 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.

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References

  1. 1.
    Levine, A.J., Momand, J. and Finlay, C.A. (1991) Nature 351, 453–456.PubMedCrossRefGoogle Scholar
  2. 2.
    Michalovitz, D., Halevy, O. and Oren, M. (1991) J. Cell. Biochem. 45, 22–29.PubMedCrossRefGoogle Scholar
  3. 3.
    Vogelstein, B. (1990) Nature 348, 681–682.PubMedCrossRefGoogle Scholar
  4. 4.
    Friedman, P.N., Kern, S.E., Vogelstein, B. and Prives, C. (1990) Proc. Nat. Acad. Sci. U.S.A. 87, 9275–9279.CrossRefGoogle Scholar
  5. 5.
    Tack, L.C., Wright, J.H., Deb, S.P. and Tegtmeyer, P. (1988) J. Virol. 63, 1310–1317.Google Scholar
  6. 6.
    Wang, E.H., Friedman, P.N. and Prives, C. (1989) Cell 57, 379–392.PubMedCrossRefGoogle Scholar
  7. 7.
    Loeber, G., Stenger, J.E., Ray, S., Parsons, R.E., Anderson, M.E. and Tegtmeyer, P. (1991) J. Virol. 65, 3167–3174.PubMedGoogle Scholar
  8. 8.
    Parsons, R.E., Stenger, J.E., Ray, S., Welker, R., Anderson, M.E. and Tegtmeyer, P. (1991) J. Virol. 65, 2798–2806.PubMedGoogle Scholar
  9. 9.
    Borowiec, J.A. and Hurwitz, J. (1988) EMBO J. 7, 3149–3158.PubMedGoogle Scholar
  10. 10.
    Dean, F.B., Bullock, P., Murakami, Y., Wobbe, R., Weissbach, L. and Hurwitz, J. (1987) Proc. Nat. Acad. Sci. U.S.A. 84, 16–20.CrossRefGoogle Scholar
  11. 11.
    Parsons, R., Anderson, M.E. and Tegtmeyer, P. (1990) J. Virol. 64, 509–518.PubMedGoogle Scholar
  12. 12.
    Stahl, H., Droege, P. and Knippers, R. (1986) EMBO J. 5, 1939–1944.PubMedGoogle Scholar
  13. 13.
    Wiekowski, M., Droge, P. and Stahl, H. (1987) J. Virol. 61, 411–418.PubMedGoogle Scholar
  14. 14.
    Wold, M.S., Li, J.J. and Kelly, T.J. (1987) Proc. Nat. Acad. Sci. U.S.A. 84, 3643–3647.CrossRefGoogle Scholar
  15. 15.
    Dornreiter, I., Höss, A., Arthur, A.K. and Fanning, E. (1990) EMBO J. 9, 3329–3336.PubMedGoogle Scholar
  16. 16.
    Smale, S.T. and Tjian, R. (1986) Mol. Cell. Biol. 6, 4077–4087.PubMedGoogle Scholar
  17. 17.
    DeCaprio, J.A., Ludlow, J.W., Figge, J., Shew, J.-Y., Huang, C.-M, Lee, W.-H., Marsilio, E., Paucha, E. and Livingston, D.M. (1988) Cell 54, 275–283.CrossRefGoogle Scholar
  18. 18.
    Lane, D.P. and Crawford, L.V. (1979) Nature 278, 261–263.PubMedCrossRefGoogle Scholar
  19. 19.
    Arthur, A.K., Höss, A. and Fanning, E. (1988) J. Virol. 62, 1999–2006.PubMedGoogle Scholar
  20. 20.
    Paucha, E., Kalderon, D., Harvey, R.W. and Smith, A.E. (1986) J. Virol. 57, 50–64.PubMedGoogle Scholar
  21. 21.
    Simmons, D.T. (1986) J. Virol. 57, 776–785.PubMedGoogle Scholar
  22. 22.
    Strauss, M., Argani, P., Mohr, I.J. and Gluzman, Y. (1987) J. Virol. 61, 3326–3330.PubMedGoogle Scholar
  23. 23.
    Klug, A. and Rhodes, D. (1987) Trends Biochem. Sci. 12, 464–469.CrossRefGoogle Scholar
  24. 24.
    Höss, A., Moarefi, I.F., Fanning, E. and Arthur, A.K. (1990) J. Virol. 64, 6291–6296.PubMedGoogle Scholar
  25. 25.
    Loeber, G., Parsons, R. and Tegtmeyer, P. (1989) J. Virol. 63, 94–100.PubMedGoogle Scholar
  26. 26.
    Berg, J.M. (1987) Cold Spring Harbor Symp. Quant. Biol. 52, 579–585.PubMedCrossRefGoogle Scholar
  27. 27.
    Berg, J.M. (1989) Cell 57, 1065–1068.PubMedCrossRefGoogle Scholar
  28. 28.
    Frankle, A.D. and Pabo, C.O. (1988) Cell 53, 675.CrossRefGoogle Scholar
  29. 29.
    Giedroc, D.P., Keating, K.P., Williams, K.R., Konigsberg, W.H. and Coleman, J.E. (1986) Proc. Nat. Acad. Sci. U.S.A. 83, 8452–8456.CrossRefGoogle Scholar
  30. 30.
    Green, S., Kumar, V., Theulaz, I., Wahli, W. and Chambon, P. (1988) EMBO J. 7, 3037–3044.PubMedGoogle Scholar
  31. 31.
    Struhl, K. (1989) Trends Biochem. Sci. 14, 137–140.PubMedCrossRefGoogle Scholar
  32. 32.
    Simmons, D.T., Wun-kim, K. and Young, W. (1990) J. Virol. 64, 4858–4865.PubMedGoogle Scholar
  33. 33.
    Finlay, C.A., Hinds, P.W. and Levine, A.J. (1989) Cell 57, 1083–1093.PubMedCrossRefGoogle Scholar
  34. 34.
    Herskowitz, I. (1987) Nature 329, 219–222.PubMedCrossRefGoogle Scholar
  35. 35.
    Fields, S. and Jang, S.K. (1990) Science 249, 1046–1049.PubMedCrossRefGoogle Scholar
  36. 36.
    O’Rourke, R.W., Miller, C.W., Kato, G.J., Simon, K.J., Chen, D.-L., Dang, S.W. and Koeffler, H.P. (1990) Oncogene 5, 1829–1832.Google Scholar
  37. 37.
    Raycroft, L., Wu, H. and Lozano, G. (1990) Science 249, 1049–1051.PubMedCrossRefGoogle Scholar
  38. 38.
    Bargonetti, J., Friedman, P.N., Kern, S.E., Vogelstein, B. and Prives, C. (1991) Cell 65, 1083–1091.PubMedCrossRefGoogle Scholar
  39. 39.
    Kern, S.E., Kinzler, K.W., Bruskin, A., Jarosz, D., Friedman, P., Prives, C. and Vogelstein, B. (1991) Science 252, 1708–1711.PubMedCrossRefGoogle Scholar
  40. 40.
    Jenkins, J.R., Chumakov, P., Addison, C., Sturzbecher, H.-W., and Wade-Evans, A. (1988) J. Virol. 62, 3906–3909.Google Scholar
  41. 41.
    Tan, T.-H., Wallis, J. and Levine, A.J. (1986) J. Virol. 59, 574–583.PubMedGoogle Scholar
  42. 42.
    Kraiss, S., Quaiser, A., Oren, M. and Montenarh, M. (1988) J. Virol. 62, 4737–4744.PubMedGoogle Scholar
  43. 43.
    McCormick, F., Clark, R., Harlow, E. and Tjian, R. (1981) Nature 292, 63–69.PubMedCrossRefGoogle Scholar
  44. 44.
    Montenarh, M. and Quaiser, A. (1989) Oncogene 4, 379–382.PubMedGoogle Scholar
  45. 45.
    Milner, J., Medcalf, E.A. and Cook, A.C. (1991) Mol. Cell. Biol. 11, 12–19.PubMedGoogle Scholar
  46. 46.
    Schmieg, F.I. and Simmons, D.T. (1988) J. Virol. 164, 132–140.CrossRefGoogle Scholar
  47. 47.
    Stenger, J.E., Mayr, G.A., Mann, K. and Tegtmeyer, P. Mol. Carcinogenesis (in press).Google Scholar
  48. 48.
    Martinez, J., Georgoff, I., Martinez, J. and Levine, A.J. (1991) Genes Develop. 5, 151–159.PubMedCrossRefGoogle Scholar
  49. 49.
    Michalovitz, D., Halevy, O. and Oren, M. (1990) Cell 62, 671–680.PubMedCrossRefGoogle Scholar
  50. 50.
    Lane, D.P. and Gannon, J. (1983) Cell Biol. Int. Rep. 7, 513–514.CrossRefGoogle Scholar
  51. 51.
    Steinmeyer, K. and Deppert, W. (1988) Oncogene 3, 501–507.PubMedGoogle Scholar
  52. 52.
    Jaenicke, R., Muiznieks, I., Aslanidis, C. and Schmitt, R. (1990) FEBS Lett. 260, 233–235.PubMedCrossRefGoogle Scholar
  53. 53.
    Kadonaga, J.T., Carner, K.R., Masiarz, F.R. and Tjian, R. (1987) Cell 51, 1079–1090.PubMedCrossRefGoogle Scholar
  54. 54.
    Mastrangelo, I.A., Courey, S.J., Wall, J.S., Jackson, S.P. and Hough, P.V.C. (1991) Proc. Nat. Acad. Sci. U.S.A. 88, 5670–5674.CrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media New York 1992

Authors and Affiliations

  • Judith E. Stenger
    • 1
  • Gregory A. Mayr
    • 1
  • Kristine Mann
    • 1
  • Satyajit Ray
    • 1
  • Mary E. Anderson
    • 1
  • Peter Tegtmeyer
    • 1
  1. 1.Department of MicrobiologyState University of New York Stony BrookUSA

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