Nucleic Acids Research, Vol 25, Issue 19 3760-3766, Copyright © 1997 by Oxford University Press
KJ Young, F Gill and JA Grasby
The hairpin ribozyme is an example of a small catalytic RNA which catalyses
the endonucleolytic transesterification of RNA in a highly
sequence-specific manner. The hairpin ribozyme, in common with all other
small ribozymes such as the hammerhead, requires the presence of a divalent
metal ion co-factor (typically magnesium) for the reaction to take place.
To investigate the role of magnesium ions in the hairpin catalysed reaction
we have synthesised two epimeric modified substrates in which a
phosphorothioate replaces the scissile phosphodiester bond. Previously,
Burke and co-workers have reported that no thio-effect is observed with the
Rp-phosphorothioate isomer. We observe the absence of a thio-effect with
both diastereomeric phosphorothioate hairpin substrates. Furthermore we
report that inert cobalt (III) complexes are capable of supporting the
hairpin ribozyme reaction, with a similar efficiency to Mg2+,even in the
presence of EDTA. Variation of the net charge on the inert cobalt complex
does not change the observed rate of reaction. These results suggest that
metal ions play a passive role in the hairpin ribozyme catalysed reaction
and are probably required for structural purposes only. This places the
hairpin ribozyme in a different mechanistic class to other small ribozymes
such as the hammerhead.
ARTICLES
Metal ions play a passive role in the hairpin ribozyme catalysed reaction
Department of Chemistry, Krebs Institute, University of Sheffield, Sheffield S3 7HF, UK.
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