Abstract
THE RNA of viroids and virusoids in plants, and the RNA tran-scripts of some tandemly repeated DNA sequences in the newt, can undergo self-catalysed cleavage to generate RNA with 5'′OH and 2′,3′-cyclic-phosphate termini1–4. These catalytic RNAs, or ribozymes, form a stem-loop secondary structure called a 'ham-merhead' in which the catalytic (ribozyme) and substrate sequences are brought close together (Fig. la). Catalytically active mimics of hammerhead ribozymes can be readily made using oligoribo-nucleotides5–7. Consequently, hammerhead analogues in which certain ribonucleotides are replaced by different ones have been constructed both to identify consensus residues required for cleavage activity and to determine the details of the cleavage mechanism8–10. But these ribonucleotide-replacements tend to alter the conformation of the hammerhead by changing hydrogen-bonding and stacking potential at the position of substitition. We have now constructed structurally less-disrupted hammerhead analogues in which deoxyribonucleotides, which lack 2′-OH groups, are substituted for ribonucleotides. These mixed RNA-DNA polymers11 were synthesized using a strategy for the chemical synthesis of RNA that is compatible with DNA synthesis12. Analy-sis of the cleavage products of several of these hammerhead analogues confirms the involvement in the reaction of the 2′-OH adjacent to the cleavage site in the substrate, and demonstrates that some 2′-OH groups in the catalytic region strongly affect activity. The results also indicate that the three-dimensional struc-ture producing nucleic acid-type catalysis is not restricted to RNA.
This is a preview of subscription content, access via your institution
Access options
Subscribe to this journal
Receive 51 print issues and online access
$199.00 per year
only $3.90 per issue
Buy this article
- Purchase on Springer Link
- Instant access to full article PDF
Prices may be subject to local taxes which are calculated during checkout
Similar content being viewed by others
References
Buzayan, J. M., Gerlach, W. L. & Bruening, G. B. Proc. natn. Acad. Sci. U.S.A. 83, 8859–8862 (1986).
Hutchins, C. J., Rathjen, P. D., Forster, A. C. & Symons, R. H. Nucleic Acids Res. 14, 3627–3640 (1986).
Forster, A. C. & Symons, R. H. Cell 49, 211–220 (1987).
Epstein, L. M. & Gall, J. G. Cell 48, 535–543 (1987).
Uhlenbeck, O. C. Nature 328, 596–600 (1987).
Jeffries, A. C. & Symons, R. H. Nucleic Acids Res. 17, 1371–1377 (1989).
Haseloff, J. & Gerlach, W. L. Nature 334, 585–591 (1988).
Sampson, J. R., Sullivan, F. X., Behlen, L. S., DiRenzo, A. B. & Uhlenbeck, O. C. Cold Spring Harb. Symp. quant. Biol. 52, 267–275 (1987).
Koizumi, M., Iwai, S. & Ohtsuka, E. FEBS Lett. 228, 228–230 (1988).
Sheldon, C. C. & Symons, R. H. Nucleic Acids Res. 17, 5679–5685 (1989).
Wu, T., Ogilvie, K. K., Perreault, J. P. & Cedergren R. J. Am. chem. Soc. 111, 8531–8533 (1989).
Usman, N., Ogilvie, K. K., Jiang, M. Y. & Cedergren, R. J. Am. chem. Soc. 109, 7845–7854 (1987).
Wu, T., Ogilvie, K. K. & Pon, R. T. Nucleic Acids Res. 17, 3501–3517 (1989).
Cedergren, R., Lang, B. F. & Gravel, D. FEBS Lett. 226, 63–66 (1987).
Zaug, A. J., Been, M. D. & Cech, T. Nature 324, 429–433 (1986).
Guerrier-Takada, C., Gardiner, K., Marsh, T., Pace, N. & Altman, S. Cell 35, 849–857 (1983).
Brown, R. S., Dewan, J. C. & Klug, A. Biochemistry 24, 4785–4801 (1985).
Koizumi, M., Hayase, Y., Iwai, S., Kamiya, H., Inoue, H. & Ohtsuka, E. Nucleic Acids Res. 17, 7059–7070 (1989).
Dickerson, R. E. J. molec. Biol. 166, 419–441 (1983).
Dock-Bregeon, A. C. et al. Nature 335, 375–378 (1988).
Chou, S.-H., Flynn, P. & Reid, B. Biochemistry 28, 2435–2443 (1989).
Paquette, J., Nicoghosian, K., Qi, G.-R., Beauchemin, N. & Cedergren, R. Eur. J. Biochem. (in the press).
Saenger, W. Principles of Nucleic Acid Structure, 331–344 (Springer, New York, 1984).
van den Hoogen, Y. et al. Eur. J. Biochem. 173, 295–303 (1988).
Joyce, G. F. Nature 338, 217–224 (1989).
Jack, A., Ladner, J. E., Rhodes, D., Brown, R. S. & Klug, A. J. molec. Biol. 111, 315–328 (1977).
Mei, H.-Y., Kaaret, T. W. & Bruice, T. C. Proc. natn. Acad. Sci. U.S.A. 86, 9727–9731 (1989).
Orgel, L. E. Cold Spring Harb. Symp. quant. Biol. 52, 9–16 (1987).
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Perreault, JP., Wu, T., Cousineau, B. et al. Mixed deoxyribo- and ribo-oligonucleotides with catalytic activity. Nature 344, 565–567 (1990). https://doi.org/10.1038/344565a0
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1038/344565a0
This article is cited by
-
Structure and mechanism of a methyltransferase ribozyme
Nature Chemical Biology (2022)
-
Crystal structure and mechanistic investigation of the twister ribozyme
Nature Chemical Biology (2014)
-
Therapeutic potential of siRNA and DNAzymes in cancer
Tumor Biology (2014)
-
Ribozyme cleavage of Plasmodium falciparum gyrase A gene transcript affects the parasite growth
Parasitology Research (2008)
-
Antiviral ribozymes
Molecular Biotechnology (1997)
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.