Abstract
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.
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
Zaug, A. J. & Cech, T. R. Science 231, 470–475 (1986).
Grosshans, C. A. & Cech, T. R. Biochemistry 28, 6888–6894 (1989).
Pyle, A. M., McSwiggen, J. A. & Cech, T. R. Proc. natn. Acad. Sci. U.S.A. 87, 8187–8191 (1990).
Celander, D. W. & Cech, T. R. Science 251, 401–407 (1991).
Piccirilli, J. A., Vyle, J. S., Caruthers, M. H. & Cech, T. R. Nature 361, 85–88 (1993).
Joyce, G. F. Gene 82, 83–87 (1989).
Beaudry, A. A. & Joyce, G. F. Science 257, 635–641 (1992).
Davies, R. W., Waring, R. B., Ray, J. A., Brown, T. A. & Scazzocchio, C. Nature 300, 719–724 (1982).
Michel, F., Jacquier, A. & Dujon, B. Biochimie 64, 867–881 (1982).
Michel, F. & Dujon, B. EMBO J. 2, 33–38 (1983).
Cech, T. R. Gene 73, 259–271 (1988).
Michel, F. & Westhof, E. J. molec. Biol. 216, 585–610 (1990).
Price, J. V., Kieft, G. L., Kent, J. R., Sievers, E. L. & Cech, T. R. Nucleic Acids Res. 13, 1871–1889 (1985).
Szostak, J. W. Nature 322, 83–86 (1986).
Burke, J. M. Gene 73, 273–294 (1988).
Couture, S. et al. J. molec. Biol. 215, 345–358 (1990).
Beaudry, A. A. & Joyce, G. F. Biochemistry 29, 6534–6539 (1990).
Herschlag, D. & Cech, T. R. Biochemistry 29, 10159–10171 (1990).
Fisher, R. A. The Genetical Theory of Natural Selection (Clarendon, Oxford, 1930).
Guatelli, J. C. et al. Proc. natn. Acad. Sci. U.S.A. 87, 1874–1878 (1990).
Joyce, G. F. in Antisense RNA and DNA (ed. Murray, J. A. H.) 353–372 (Wiley-Liss, New York, 1992).
Milligan, J. F., Groebe, D. R., Witherell, G. W. & Uhlenbeck, O. C. Nucleic Acids Res. 15, 8783–8798 (1987).
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Lehman, N., Joyce, G. Evolution in vitro of an RNA enzyme with altered metal dependence. Nature 361, 182–185 (1993). https://doi.org/10.1038/361182a0
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1038/361182a0
This article is cited by
-
Evolution of the Standard Genetic Code
Journal of Molecular Evolution (2021)
-
Construction and characterization of an improved DNA-launched infectious clone of duck hepatitis a virus type 1
Virology Journal (2017)
-
Empirical demonstration of environmental sensing in catalytic RNA: evolution of interpretive behavior at the origins of life
BMC Evolutionary Biology (2014)
-
The Effects of Stabilizing and Directional Selection on Phenotypic and Genotypic Variation in a Population of RNA Enzymes
Journal of Molecular Evolution (2014)
-
HAPIscreen, a method for high-throughput aptamer identification
Journal of Nanobiotechnology (2011)
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.