Letter | Published:

Novel guanosine requirement for catalysis by the hairpin ribozyme

Nature volume 354, pages 320322 (28 November 1991) | Download Citation

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Abstract

THERE is much interest in the development of 'designer ribozymes' to target destruction of RNAs in vitro and in vivo1. Engineering of ribozymes with novel specificities requires detailed knowledge of the ribozyme-substrate interaction, and a rigorous evaluation of sequence specificity. The hairpin ribozyme catalyses an efficient and reversible site-specific cleavage reaction2–4. We have used mutagenesis and in vitro selection strategies to show that RNA cleavage and ligation has an absolute requirement for guanosine immediately 3' to the cleavage-ligation site. This G is not required for efficient substrate binding, rather, its 2-amino group is an essential component of the active site required for catalysis.

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Author information

Affiliations

  1. Department of Microbiology and Molecular Genetics, Markey Center for Molecular Genetics, The University of Vermont, Burlington, Vermont 05405, USA

    • Bharat M. Chowrira
    • , Alfredo Berzal-Herranz
    •  & John M. Burke

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DOI

https://doi.org/10.1038/354320a0

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