A re‐investigation of the thio effect at the hammerhead cleavage site

Author： Scott Edmund C.   Uhlenbeck Olke C.  

Publisher： Oxford University Press

ISSN： 1362-4962

Source： Nucleic Acids Research, Vol.27, Iss.2, 1999-01, pp. : 479-484

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Abstract

The effect of introducing a phosphorothioate at the hammerhead cleavage site was investigated using a kinetically well‐characterized hammerhead. In buffers containing Mg ion, the RP‐phosphorothioate isomer cleaved 2000‐ to 80 000‐fold slower than the SP isomer or the unmodified RNA substrate. Addition of low concentrations of several thiophilic metal ions, especially Cd2+, to these reactions is sufficient to fully restore the cleavage rate of the RP substrate without affecting cleavage rate of the all‐oxygen or SP substrate. Thus, a model proposing coordination of a divalent metal ion to the pro‐R oxygen at the hammerhead cleavage site appears justified.

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