Sequence-specific cleavage of oligoribonucleotide capable of forming a stem and loop structure

Abstract

The precursor of an RNA molecule from T4-infected Escherichia coli cells (p2Sp1 RNA) has the capacity to cleave itself at specific positions ((UpA (137-138) and CpA (170-171)) within a possible loop and stem structure. This specific cleavage requires at least a monovalent cation and nonionic detergents. To confirm that the sequence-specific cleavage occurs autolytically, we studied the selective hydrolysis of RNA using a chemically synthesized 13-mer (GUUUCGUACAAAC) having sequences corresponding to G131-C143 of p2Sp1 RNA. The cleavage occurred at two identical sites (UpA and CpA) of a hairpin loop under physiological conditions and was affected by monovalent or divalent metals, nonionic detergent, salt, pH, and temperature. The hairpin loop domain and specific sequences are necessary for the cleavage of RNA 13-mer (GUUUCGUACAAAC).