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Science 1 October 1999:
Vol. 286. no. 5437, pp. 123 - 126
DOI: 10.1126/science.286.5437.123
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Reports

Imidazole Rescue of a Cytosine Mutation in a Self-Cleaving Ribozyme

Anne T. Perrotta, I-hung Shih, Michael D. Been *

Ribozymes use a number of the same catalytic strategies as protein enzymes. However, general base catalysis by a ribozyme has not been demonstrated. In the hepatitis delta virus antigenomic ribozyme, imidazole buffer rescued activity of a mutant with a cytosine-76 (C76) to uracil substitution. In addition, a C76 to adenine substitution reduced the apparent pKa (where Ka is the acid constant) of the self-cleavage reaction by an amount consistent with differences in the pKa values of these two side chains. These results suggest that, in the wild-type ribozyme, C76 acts as a general base. This finding has implications for potential catalytic functions of conserved cytosines and adenines in other ribozymes and in ribonuclear proteins with enzymatic activity.

Department of Biochemistry, Box 3711, Duke University Medical Center, Durham, NC 27710 USA.
*   To whom correspondence should be addressed. E-mail: been{at}biochem.duke.edu

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