Skip to main content

Thank you for visiting nature.com. You are using a browser version with limited support for CSS. To obtain the best experience, we recommend you use a more up to date browser (or turn off compatibility mode in Internet Explorer). In the meantime, to ensure continued support, we are displaying the site without styles and JavaScript.

  • Article
  • Published:

Changing the binding specificity of a represser by redesigning an α-helix

Nature volume 316, pages 601–605 (1985)Cite this article

Abstract

We replaced amino acids on the ‘outside’, or solvent-exposed, surface of the DNA recognition α-helix of 434 repressor with the corresponding amino acids from the recognition helix of P22 repressor. The binding specificity of the resulting hybrid protein, as measured in vivo and in vitro, was that of P22 repressor.

This is a preview of subscription content, access via your institution

Access options

Buy this article

  • Purchase on Springer Link
  • Instant access to full article PDF
Buy now

Prices may be subject to local taxes which are calculated during checkout

Similar content being viewed by others

References

  1. Anderson, J. E., Ptashne, M. & Harrison, S. C. Nature 316, 596–601 (1985).

    Article  ADS  CAS  Google Scholar 

  2. Pabo, C. O. & Lewis, M. Nature 298, 443–447 (1982).

    Article  ADS  CAS  Google Scholar 

  3. Anderson, W. F., Ohlendorf, D. H., Takeda, Y. & Matthews, B. W. Nature 290, 754–758 (1981).

    Article  ADS  CAS  Google Scholar 

  4. McKay, D. B. & Steitz, T. A. Nature 290, 744–749 (1981).

    Article  ADS  CAS  Google Scholar 

  5. Steitz, T. A., Ohlendorf, D. H., McKay, D. B., Anderson, W. F. & Matthews, B. W. Proc. natn. Acad. Sci. U.S.A. 79, 3097–3100 (1982).

    Article  ADS  CAS  Google Scholar 

  6. Ohlendorf, D. H., Anderson, W. F., Lewis, M., Pabo, C. O. & Matthews, B. W. J. molec. Biol. 169, 757–769 (1983).

    Article  CAS  Google Scholar 

  7. Lewis, M. et al. Cold Spring Harb. Symp. quant. Biol. 47, 435–440 (1983).

    Article  Google Scholar 

  8. Ohlendorf, D. H., Anderson, W. F., Fisher, R. G., Takeda, Y. & Matthews, B. W. Nature 298, 718–723 (1982).

    Article  ADS  CAS  Google Scholar 

  9. Weber, I. T. & Steitz, T. A. Proc. natn. Acad. Sci. U.S.A. 81, 3973–3977 (1984).

    Article  ADS  CAS  Google Scholar 

  10. Sauer, R. T., Yocum, R. R., Doolittle, R. F., Lewis, M. & Pabo, C. O. Nature 298, 447–451 (1982).

    Article  ADS  CAS  Google Scholar 

  11. Anderson, W. D., Takeda, Y., Ohlendorf, D. H. & Matthews, B. W. J molec. Biol. 159, 745–751 (1982).

    Article  CAS  Google Scholar 

  12. Weber, I. T., McKay, D. B. & Steitz, T. A. Nucleic Acids Res. 10, 5085–5102 (1982).

    Article  CAS  Google Scholar 

  13. Wharton, R. P., Brown, E. L. & Ptashne, M. Cell 38, 361–369 (1984).

    Article  CAS  Google Scholar 

  14. Hecht, M. H., Nelson, H. C. M. & Sauer, R. T. Proc. natn. Acad. Sci. U.S.A. 80, 2676–2680 (1983).

    Article  ADS  CAS  Google Scholar 

  15. Pabo, C. O. & Sauer, R. T. A. Rev. Biochem. 53, 293–321 (1984).

    Article  CAS  Google Scholar 

  16. Ebright, R. H., Cossart, P., Gicquel-Sanzey, B. & Beckwith, J. Nature 311, 232–235 (1984).

    Article  ADS  CAS  Google Scholar 

  17. Kelley, R. L. & Yanofsky, C. Proc. natn. Acad. Sci. U.S.A. 82, 483–487 (1985).

    Article  ADS  CAS  Google Scholar 

  18. Eisenbis, S. et al. Proc. natn. Acad. Sci. U.S.A. 82, 1084–1088 (1985).

    Article  ADS  Google Scholar 

  19. Kaiser, A. D. & Jacob, F. Virology 4, 509–512 (1957).

    Article  CAS  Google Scholar 

  20. Botstein, D. & Herskowitz, I. Nature 251, 584–589 (1974).

    Article  ADS  CAS  Google Scholar 

  21. Anderson, J. E. thesis, Harvard Univ. (1984).

  22. De Anda, J., Poteete, A. R. & Sauer, R. T. J. biol Chem. 258, 10536–10542 (1983).

    CAS  PubMed  Google Scholar 

  23. Poteete, A. R., Ptashne, M., Ballivet, M. & Eisen, H. J. molec. Biol. 137, 81–91 (1980).

    Article  CAS  Google Scholar 

  24. Poteete, A. R. & Ptashne, M. J. molec. Biol. 157, 21–48 (1982).

    Article  CAS  Google Scholar 

  25. Nelson, H. C. M. & Sauer, R. T. Cell (in the press).

  26. Coulondre, C. & Miller, J. H. J molec. Biol. 117, 525–575 (1977).

    Article  CAS  Google Scholar 

  27. Calos, M. P. & Miller, J. H. Mol. gen. Genet. 183, 559–560 (1981).

    Article  CAS  Google Scholar 

  28. Laemmli, U. Nature 227, 680–682 (1970).

    Article  ADS  CAS  Google Scholar 

  29. Poteete, A. R. & Roberts, T. M. Gene 13, 153–161 (1981).

    Article  CAS  Google Scholar 

  30. Hochschild, A., Irwin, N. & Ptashne, M. Cell 32, 319–325 (1983).

    Article  CAS  Google Scholar 

  31. Schiffer, M. & Edmundson, A. B. Biophys. J. 7, 121–135 (1967).

    Article  CAS  Google Scholar 

  32. Zoller, M. J. & Smith, M. Meth. Enzym. 100, 468–500 (1983).

    Article  CAS  Google Scholar 

  33. Dente, L., Cesareni, G. & Cortese, R. Nucleic Acids Res. 11, 1645–1655 (1983).

    Article  CAS  Google Scholar 

  34. Sgaramella, V. & Khorana, H. G. J. molec. Biol. 72, 427–444 (1972).

    Article  CAS  Google Scholar 

  35. Riggs, A. D., Suzuki, H. & Bourgeois, S. J. molec. Biol. 48, 67–83 (1970).

    Article  CAS  Google Scholar 

  36. Anderson, J. E., Ptashne, M. & Harrison, S. C. Proc. natn. Acad. Sci. U.S.A. 81, 1307–1311 (1984).

    Article  ADS  CAS  Google Scholar 

  37. Maniatis, T., Fritsch, E. F. & Sambrook, J. Molecular Cloning, A Laboratory Manual (Cold Spring Harbor Laboratory, New York, 1982).

    Google Scholar 

  38. Sanger, F., Coulson, A. R., Barrell, B. G., Smith, A. J. H. & Roe, B. A. J. molec. Biol. 143, 161–178 (1980).

    Article  CAS  Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Department of Biochemistry and Molecular Biology, Harvard University, 7 Divinity Avenue, Cambridge, Massachusetts, 02138, USA

    Robin P. Wharton & Mark Ptashne

Authors
  1. Robin P. Wharton
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Mark Ptashne
    View author publications

    You can also search for this author in PubMed Google Scholar

Rights and permissions

Reprints and permissions

About this article

Cite this article

Wharton, R., Ptashne, M. Changing the binding specificity of a represser by redesigning an α-helix. Nature 316, 601–605 (1985). https://doi.org/10.1038/316601a0

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1038/316601a0

This article is cited by

Comments

By submitting a comment you agree to abide by our Terms and Community Guidelines. If you find something abusive or that does not comply with our terms or guidelines please flag it as inappropriate.

Search

Advanced search

Quick links

Nature Briefing

Sign up for the Nature Briefing newsletter — what matters in science, free to your inbox daily.

Get the most important science stories of the day, free in your inbox. Sign up for Nature Briefing