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.

  • Letter
  • Published:

A DNA metalloenzyme with DNA ligase activity

Abstract

SINGLE-STRANDED DNA can fold into well-defined sequence-dependent tertiary structures that specifically bind a variety of target molecules1–10, raising the possibility that some folded single-stranded DNAs might exhibit catalytic activities similar to those of ribozymes and protein enzymes. Derivatives of the hammerhead ribozyme that contain a majority of deoxyribonucleotides retain the ability to cleave RNA11, and a 'deoxyribozyme' was generated by leaving all essential ribonucleotides of the hammerhead on the RNA 'substrate'12. Recently in vitro selection has been used to isolate a DNA sequence that shows Pb2+-dependent RNA-cleaving activity13. Here we report the isolation by in vitro selection14–17 of a small single-stranded DNA that is a Zn2+/Cu2 +-dependent metalloenzyme. The enzyme catalyses the formation of a new phos-phodiester bond by the condensation of the 5′–hydroxyl of one oligodeoxynucleotide and a 3′-phosphorimidazolide on another oli-godeoxynucleotide, and shows multiple turnover ligation.

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

Access options

Rent or buy this article

Prices vary by article type

from$1.95

to$39.95

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

Similar content being viewed by others

References

  1. Khan, A. S. & Roe, B. A. Science 241, 74–76 (1988).

    Article  ADS  CAS  Google Scholar 

  2. Perreault, J.-P. et al. Eur. J. Biochem. 186, 87–93 (1989).

    Article  CAS  Google Scholar 

  3. Bock, L. C., Griffin, L. C., Latham, J. A., Vermaas, E. H. & Toole, J. J. Nature 355, 564–566 (1992).

    Article  ADS  CAS  Google Scholar 

  4. Ellington, A. D. & Szostak, J. W. Nature 355, 850–852 (1992).

    Article  ADS  CAS  Google Scholar 

  5. Blackburn, E. H. A. Rev. Biochem. 61, 113–129 (1992).

    Article  CAS  Google Scholar 

  6. Wang, K. Y., Krawczyk, S. H., Bischofberger, N., Swaminathan, S. & Bolton, P. H. Biochemistry 32, 11285–11292 (1993).

    Article  CAS  Google Scholar 

  7. Paborsky, L. R., McCurdy, S. N., Griffin, L. C., Toole, J. J. & Leung, L. L. K. J. biol. Chem. 268, 20808–20811 (1993).

    CAS  PubMed  Google Scholar 

  8. Padmanabhan, K., Padmanabhan, K. P., Ferrara, J. D., Sadler, J. E. & Tulinsky, A. J. biol. Chem. 268, 17651–17654 (1993).

    CAS  PubMed  Google Scholar 

  9. Wyatt, J. R. et al. Proc. natn. Acad. Sci. U.S.A. 91, 1356–1360 (1994).

    Article  ADS  CAS  Google Scholar 

  10. Huizenga, D. E. & Szostak, J. W. Biochemistry 34, 656–665 (1995).

    Article  CAS  Google Scholar 

  11. Bratty, J., Chartrand, P., Ferbeyre, G. & Cedergren, R. Biochim. biophys. Acta 1216, 345–359 (1993).

    Article  CAS  Google Scholar 

  12. Chartrand, P., Harvey, S. C., Ferbeyre, G., Usman, N. & Cedergren, R. in RNA Processing 77 (Proc. RNA Soc., Madison, Wisconsin, 1994).

    Google Scholar 

  13. Breaker, R. R. & Joyce, G. F. Chem. Biol. 1, 223–229 (1994).

    Article  CAS  Google Scholar 

  14. Szostak, J. W. Trends biochem. Sci. 17, 89–93 (1992).

    Article  CAS  Google Scholar 

  15. Chapman, K. B. & Szostak, J. W. Curr. Opin. Struct. Biol. 4, 618–622 (1994).

    Article  CAS  Google Scholar 

  16. Breaker, R. R. & Joyce G. F. Trends Biotechnol. 12, 268–275 (1994).

    Article  CAS  Google Scholar 

  17. Joyce, G. F. Curr. Opin. struct. Biol. 4, 331–336 (1994).

    Article  CAS  Google Scholar 

  18. Shabarova, Z. A. Biochimie 70, 1323–1334 (1988).

    Article  CAS  Google Scholar 

  19. Luebke, K. J. & Dervan, P. B. J. Am. chem. Soc. 111, 8733–8735 (1989).

    Article  CAS  Google Scholar 

  20. Lerner, R. A., Benkovic, S. J. & Schultz, P. G. Science 252, 659–667 (1991).

    Article  ADS  CAS  Google Scholar 

  21. Fersht, A. Enzyme Structure and Mechanism (Freeman, New York, 1985).

    Google Scholar 

  22. Pyle, A. M. Science 261, 709–714 (1993).

    Article  ADS  CAS  Google Scholar 

  23. Karlin, K. D. Science 261, 701–708 (1993).

    Article  ADS  CAS  Google Scholar 

  24. Dahm, S. C., Derrick, W. B. & Uhlenbeck, O. C. Biochemistry 32, 13040–13045 (1993).

    Article  CAS  Google Scholar 

  25. Pan, T., Ditchl, B. & Uhlenbeck, O. C. Biochemistry 33, 9561–9565 (1994).

    Article  CAS  Google Scholar 

  26. Kao, T. H. & Crothers, D. M. Proc. natn. Acad. Sci. U.S.A. 77, 3360–3364 (1980).

    Article  ADS  CAS  Google Scholar 

  27. Orgel, L. E. Scient. Am. 271(4), 76–83 (1994).

    Article  CAS  Google Scholar 

  28. Chu, B. C. F., Kramer, F. R. & Orgel, L. E. Nucleic Acids Res. 14, 5591–5603 (1986).

    Article  CAS  Google Scholar 

  29. Wilson, C. & Szostak, J. W. Nature 374, 777–782 (1995).

    Article  ADS  CAS  Google Scholar 

  30. Bailar, J. C. Jr, Emeléus, H. J. & Trotman-Dickenson, A. F. Comprehensive Inorganic Chemistry (Pergamon, Elmsford, N.Y., 1973).

    Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Rights and permissions

Reprints and permissions

About this article

Cite this article

Cuenoud, B., Szostak, J. A DNA metalloenzyme with DNA ligase activity. Nature 375, 611–614 (1995). https://doi.org/10.1038/375611a0

Download citation

  • Received:

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1038/375611a0

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

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