Chemically Modified Hammerhead Ribozymes with Improved Catalytic Rates

Abstract

A site-specific chemical modification strategy has been employed to elucidate structure−function relationships at the only phylogenetically nonconserved position within the core of the hammerhead ribozyme (N7). Four different base substitutions at position 7 resulted in increased catalytic rates. A pyridin-4-one base substitution increased the rate of the chemical step up to 12-fold. These results are the first examples of chemical modifications within a catalytic RNA that enhance the rate of the chemical step. Four base substitutions resulted in decreased catalytic rates. The results do not correlate with proposed hydrogen bond interactions (Pley et al., 1994; Scott et al., 1995). This study demonstrates the utility of using unnatural nucleotide analogsrather than mutagenesis with the four standard nucleotides aloneto elucidate structure−function relationships of small RNAs.