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The hammerhead cleavage reaction in monovalent cations

Published online by Cambridge University Press:  09 April 2001

EDWARD A. CURTIS
Affiliation:
Whitehead Institute for Biomedical Research and Department of Biology, Massachusetts Institute of Technology, Cambridge, Massachusetts 02142, USA
DAVID P. BARTEL
Affiliation:
Whitehead Institute for Biomedical Research and Department of Biology, Massachusetts Institute of Technology, Cambridge, Massachusetts 02142, USA
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Abstract

Recently, Murray et al. (Chem Biol, 1998, 5:587–595) found that the hammerhead ribozyme does not require divalent metal ions for activity if incubated in high (≥1 M) concentrations of monovalent ions. We further characterized the hammerhead cleavage reaction in the absence of divalent metal. The hammerhead is active in a wide range of monovalent ions, and the rate enhancement in 4 M Li+ is only 20-fold less than that in 10 mM Mg2+. Among the Group I monovalent metals, rate correlates in a log-linear manner with ionic radius. The pH dependence of the reaction is similar in 10 mM Mg2+, 4 M Li+, and 4 M Na+. The exchange-inert metal complex Co(NH3)63+ also supports substantial hammerhead activity. These results suggest that a metal ion does not act as a base in the reaction, and that the effects of different metal ions on hammerhead cleavage rates primarily reflect structural contributions to catalysis.

Type
Research Article
Copyright
© 2001 RNA Society

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