Complete identification of nonbridging phosphate oxygens involved in hammerhead cleavage.

摘要: Phosphorothioate interference analysis is suited for the rapid identification of structurally and functionally important phosphate groups. Previous studies, however, have been limited to pro-Rp oxygens. We employed solid-phase oligonucleotide synthesis modification investigate either nonbridging oxygens within hammerhead ribozyme. Two novel sites Sp phosphorothioate were identified at positions A6 U16.1. The results from experiments confirmed by single substitutions interest. Metal rescue revealed that direct metal ion coordination occurs in functional ribozyme only site cleavage oxygen position Ag. new approach may be generally useful rapidly evaluating importance groups nucleic acids.