Nucleic Acids Research, 1991, Vol. 19, No. 6 1183-1188
© 1991
CHEMISTRY |
Configurationally defined phosphorothioate-containing oligoribonucleotides in the study of the mechanism of cleavage of hammerhead ribozymes
MRC Laboratory of Molecular Biology Hills Road, Cambridge CB2 2QH, UK
* To whom correspondence should be addressed
Received January 31, 1990. Accepted February 21, 1991.
The chemical synthesis is described of oligoribonucleotides containing a single phosphoro-thioate linkage of defined Rp and Sp configuration. The oligoribonucleotides were used as substrates in the study of the mechanism of cleavage of an RNA hammerhead domain having the phosphorothioate group at the cleavage site. Whereas the Rp Isomer was cleaved only very slowly in the presence of magnesium ion, the rate of cleavage of the Sp isomer was only slightly reduced from that of the unmodified phosphodiester. This finding gives further evidence for the hypothesis that the magnesium ion is bound to the pro-R oxygen in the transition state of the hammerhead cleavage reaction. Also, inversion of configuration at phosphorus is confirmed for a two-stranded hammerhead.
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