Self-cleavage of p2Sp1 RNA with Mg2+ and non-ionic detergent (Brij 58)

The precursor of an RNA molecule from T4-infected E. coli cells (p2Sp1 RNA) has the capacity to cleave itself at specific positions [(UpA (139-140) an d CpA (170-171)], within a putative loop and stem structure. This sequence- specific cleavage requires at least a monovalent cation and non-ionic deter gents. We studied the self-cleavage reaction of an RNA fragment (GUUUCGUACA AAC) (R1) with the sequence corresponding to the p2Sp1 RNA in the presence of Mg2+ and non-ionic detergents. It requires Mg2+ and is aided by a non-io nic detergent, Brij 58. The cleavage reaction is time, temperature, and pH- dependent. The cleavage occurs at the phosphodiester bond between UpA and C pA on the RNA fragment (GUUUCGUACAAAC) (R1). Furthermore, the maximum of cl eavage of R1 occurs at a very low Mg2+ concentration (less than or equal to 5 mM). (C) 2000 Elsevier Science B.V. All rights reserved.