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ATP-dependent assembly of double hexamers of SV40 T antigen at the viral origin of DNA replication

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Abstract

SIMIAN virus 40 (SV40) replicates in nuclei of human and monkey cells. One viral protein, large tumour (T) antigen, is required for the initiation of DNA replication1,2. The development of in vitro replication systems which retain this property has facilitated the identification of the cellular components required for replication3–5. T antigen recognizes the pentanucleotide 5′-GAGGC-3′ which is present in four copies6,7 within the 64 base-pairs (bp) of the core origin8,9. In the presence of ATP it binds with increased affinity10,11 forming a distinctive, bilobed structure visible in electron micrographs12. As a helicase13, it unwinds SV40 DNA bidirectional ly from the origin14–16. We report here that in vitro and in the presence of ATP, T antigen assembles a double hexamer, centred on the core origin and extending beyond it by 12 bp in each direction. The assembly of this dodecamer initiates an untwisting of the duplex by 2–3 turns. In the absence of ATP, a tetrameric structure is the largest found at the core origin. In the absence of DNA, but in the presence of ATP or its non-hydrolysable analogues, T antigen assembles into hexamers. This suggests that ATP effects an allosteric change in the monomer. The change alters protein-protein interactions and allows the assembly of a double hexamer, which initiates replication at the core origin.

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References

  1. Tegtmeyer, P. J. Virol. 10, 591–598 (1972).

    CAS  PubMed  PubMed Central  Google Scholar 

  2. Myers, R. M. & Tijan, R. Proc. natn. Acad. Sci. U.S.A. 77, 6491–6495 (1980).

    Article  ADS  CAS  Google Scholar 

  3. Li, J. J. & Kelly, T. J. Proc. natn. Acad. Sci. U.S.A. 81, 6973–6977 (1984).

    Article  ADS  CAS  Google Scholar 

  4. Stillman, B. W. & Gluzman, Y. Molec. cell. Biol. 5, 2051–2060 (1985).

    Article  CAS  Google Scholar 

  5. Wobbe, C. R., Dean, F. B., Weissbach, L. & Hurwitz, J. Proc. natn. Acad. Sci. U.S.A. 82, 5710–5714 (1985).

    Article  ADS  CAS  Google Scholar 

  6. Tjian, R. Cold Spring Harb. Symp. quant. Biol. 43, 655–662 (1978).

    Article  Google Scholar 

  7. DeLucia, A., Lewton, B., Tjian, R. & Tegtmeyer, P. J. Virol. 46, 143–150 (1983).

    CAS  PubMed  PubMed Central  Google Scholar 

  8. Deb, S., DeLucia, A., Baur, C., Koff, A. & Tegtmeyer, P. Molec. cell. Biol. 6, 1663–1670 (1986).

    Article  CAS  Google Scholar 

  9. Deb, S., DeLucia, A., Koff, A., Tsui, S. & Tegtmeyer, P. Molec. cell. Biol. 6, 4578–4584 (1986).

    Article  CAS  Google Scholar 

  10. Deb, S. P. & Tegtmeyer, P. J. Virol. 61, 3649–3654 (1987).

    CAS  PubMed  PubMed Central  Google Scholar 

  11. Borowiec, J. A. & Hurwitz, J. Proc. natn. Acad. Sci. U.S.A. 85, 64–68 (1988).

    Article  ADS  CAS  Google Scholar 

  12. Dean, F. B., Dodson, M., Echols, H. & Hurwitz, J. Proc. natn. Acad. Sci. U.S.A. 84, 8981–8985 (1987).

    Article  ADS  CAS  Google Scholar 

  13. Stahl, H., Droge, P. & Knippers, R. EMBO J. 5, 1939–1944 (1986).

    Article  CAS  Google Scholar 

  14. Dean, F. B. et al. Proc. natn. Acad. Sci. U.S.A. 84, 16–20 (1987).

    Article  ADS  CAS  Google Scholar 

  15. Wold, M. S., Li, J. J. & Kelly, T. J. Proc. natn. Acad. Sci. U.S.A. 84, 3643–3647 (1987).

    Article  ADS  CAS  Google Scholar 

  16. Dodson, M., Dean, F. B., Bullock, P., Echols, H. & Hurwitz, J. Science 238, 964–967 (1987).

    Article  ADS  CAS  Google Scholar 

  17. Wall, J. S. in Analytical Electron Microscopy (eds Hren, J., Goldstein, J. & Joy, D.) 333–342 (Plenum, New York, 1979).

    Google Scholar 

  18. Mastrangelo, I. A. et al Proc. natn. Acad. Sci. U.S.A. 82, 3626–3630 (1985).

    Article  ADS  CAS  Google Scholar 

  19. Hough, P. V. C. et al. Proc. natn. Acad. Sci. U.S.A. 84, 4826–4830 (1987).

    Article  ADS  CAS  Google Scholar 

  20. Buchman, A. R., Burnett, L. & Berg, P. in DNA Tumor Viruses (ed. Tooze, J.) 799–818 (Cold Spring Harbor Laboratory, New York, 1980).

    Google Scholar 

  21. Wall, J. S., Hainfeld, J. K., Bartlett, P. A. & Singer, S. J. J. Ultramicros. 8, 397–402 (1982).

    Article  CAS  Google Scholar 

  22. Bradley, M. K., Griffin, J. D. & Livingston, D. M. Cell 28, 125–134 (1982).

    Article  CAS  Google Scholar 

  23. Fanning, E., Westphal, D.-H., Brauer, D. & Corlin, D. EMBO J. 1, 1023–1028 (1982).

    Article  CAS  Google Scholar 

  24. Gidoni, D., Scheller, B. B., Hantzopoulos, P., Oren, M. & Prives, C. J. Virol. 42, 456–466, (1982).

    CAS  PubMed  PubMed Central  Google Scholar 

  25. Montenarh, M. & Henning, R. J. Virol. 45, 531–538 (1983).

    CAS  PubMed  PubMed Central  Google Scholar 

  26. Wobbe, C. R., Dean, F. B., Murakami, Y., Weissbach, L. & Hurwitz, J. Proc natn Acad. Sci. U.S.A. 83, 4612–4616 (1986).

    Article  ADS  CAS  Google Scholar 

  27. Arai, K. & Kornberg, A. J. biol. Chem. 256, 5260–5266 (1981).

    CAS  PubMed  Google Scholar 

  28. LeBowitz, J. H. & McMacken, R. J. biol. Chem. 261, 4738–4748 (1986).

    CAS  PubMed  Google Scholar 

  29. Liu, C.-C. & Alberts, B. M. J. biol. Chem. 256, 2813–2820 (1981).

    CAS  PubMed  Google Scholar 

  30. Reha-Krantz, L. J. & Hurwitz, J. J. biol. Chem. 253, 4043–4050 (1978).

    CAS  PubMed  Google Scholar 

  31. Finger, L. R. & Richardson, J. P. J. molec. Biol. 156, 203–219 (1982).

    Article  CAS  Google Scholar 

  32. Brennan, C. A., Dombroski, A. J. & Platt, T. Cell 48, 945–952 (1987).

    Article  CAS  Google Scholar 

  33. Borowiec, J. A. & Hurwitz, J. EMBO J. 7, 3149–3158 (1988).

    Article  CAS  Google Scholar 

  34. DeLucia, A., Deb, S., Partin, K. & Tegtmeyer, P. J. Virol. 57, 138–144 (1986).

    CAS  PubMed  PubMed Central  Google Scholar 

  35. Dixon, R. A. F. & Nathans, D. J. Virol. 53, 1001–1004 (1985).

    CAS  PubMed  PubMed Central  Google Scholar 

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Authors and Affiliations

  1. Biology Department, Brookhaven National Laboratory, Upton, New York, 11973, USA

    Iris A. Mastrangelo, Paul V. C. Hough & Joseph S. Wall

  2. Department of Biochemistry, Stanford University School of Medicine, Stanford, California, 94305, USA

    Mark Dodson

  3. Graduate Program in Molecular Biology, Sloan Kettering Memorial Institute, New York, 10021, USA

    Frank B. Dean & Jerard Hurwrtz

Authors
  1. Iris A. Mastrangelo
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  2. Paul V. C. Hough
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  3. Joseph S. Wall
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  4. Mark Dodson
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  5. Frank B. Dean
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  6. Jerard Hurwrtz
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Mastrangelo, I., Hough, P., Wall, J. et al. ATP-dependent assembly of double hexamers of SV40 T antigen at the viral origin of DNA replication. Nature 338, 658–662 (1989). https://doi.org/10.1038/338658a0

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