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Letters to Nature
Nature 361, 182 - 185 (14 January 1993); doi:10.1038/361182a0

Evolution in vitro of an RNA enzyme with altered metal dependence

Niles Lehman & Gerald F. Joyce

Departments of Chemistry and Molecular Biology, The Scripps Research Institute, 10666 North Torrey Pines Road, La Jolla, California 92037, USA

THE Tetrahymena group I ribozyme catalyses a sequence-specific phosphodiester cleavage reaction on an external RNA oligonucleotide substrate in the presence of a divalent metal cation cofactor1. This reaction proceeds readily with either Mg2+ or Mn2+, but no detectable reaction has been reported when other divalent cations are used as the sole cofactor2–5. Cations such as Ca2+, Sr2+ and Ba2+ can stabilize the correct folded conformation of the ribozyme, thereby partially alleviating the Mg2+ or Mn2+ requirement2–5. But catalysis by the ribozyme involves coordination of either Mg2+ or Mn2+ at the active site, resulting in an overall requirement for one of these two cations5. Here we use an in vitro evolution process6,7 to obtain variants of the Tetrahymena ribozyme that are capable of cleaving an RNA substrate in reaction mixtures containing Ca2+ as the divalent cation. These findings extend the range of different chemical environments available to RNA enzymes and illustrate the power of in vitro evolution in generating macromolecular catalysts with desired properties.

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References

1. Zaug, A. J. & Cech, T. R. Science 231, 470−475 (1986). | PubMed | ISI | ChemPort |
2. Grosshans, C. A. & Cech, T. R. Biochemistry 28, 6888−6894 (1989). | Article | PubMed | ISI | ChemPort |
3. Pyle, A. M., McSwiggen, J. A. & Cech, T. R. Proc. natn. Acad. Sci. U.S.A. 87, 8187−8191 (1990). | ChemPort |
4. Celander, D. W. & Cech, T. R. Science 251, 401−407 (1991). | PubMed | ISI | ChemPort |
5. Piccirilli, J. A., Vyle, J. S., Caruthers, M. H. & Cech, T. R. Nature 361, 85−88 (1993). | Article | PubMed | ISI | ChemPort |
6. Joyce, G. F. Gene 82, 83−87 (1989). | Article | PubMed | ISI | ChemPort |
7. Beaudry, A. A. & Joyce, G. F. Science 257, 635−641 (1992). | PubMed | ISI | ChemPort |
8. Davies, R. W., Waring, R. B., Ray, J. A., Brown, T. A. & Scazzocchio, C. Nature 300, 719−724 (1982). | Article | PubMed | ISI | ChemPort |
9. Michel, F., Jacquier, A. & Dujon, B. Biochimie 64, 867−881 (1982). | PubMed | ISI | ChemPort |
10. Michel, F. & Dujon, B. EMBO J. 2, 33−38 (1983). | PubMed | ISI | ChemPort |
11. Cech, T. R. Gene 73, 259−271 (1988). | Article | PubMed | ISI | ChemPort |
12. Michel, F. & Westhof, E. J. molec. Biol. 216, 585−610 (1990). | Article | PubMed | ISI | ChemPort |
13. Price, J. V., Kieft, G. L., Kent, J. R., Sievers, E. L. & Cech, T. R. Nucleic Acids Res. 13, 1871−1889 (1985). | PubMed | ChemPort |
14. Szostak, J. W. Nature 322, 83−86 (1986). | Article | PubMed | ISI | ChemPort |
15. Burke, J. M. Gene 73, 273−294 (1988). | Article | PubMed | ISI | ChemPort |
16. Couture, S. et al. J. molec. Biol. 215, 345−358 (1990). | PubMed | ISI | ChemPort |
17. Beaudry, A. A. & Joyce, G. F. Biochemistry 29, 6534−6539 (1990). | Article | PubMed | ChemPort |
18. Herschlag, D. & Cech, T. R. Biochemistry 29, 10159−10171 (1990). | Article | PubMed | ISI | ChemPort |
19. Fisher, R. A. The Genetical Theory of Natural Selection (Clarendon, Oxford, 1930).
20. Guatelli, J. C. et al. Proc. natn. Acad. Sci. U.S.A. 87, 1874−1878 (1990). | ChemPort |
21. Joyce, G. F. in Antisense RNA and DNA (ed. Murray, J. A. H.) 353−372 (Wiley-Liss, New York, 1992). | ChemPort |
22. Milligan, J. F., Groebe, D. R., Witherell, G. W. & Uhlenbeck, O. C. Nucleic Acids Res. 15, 8783−8798 (1987). | PubMed | ISI | ChemPort |



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