Rearrangement of a Stable RNA Secondary Structure during VS Ribozyme Catalysis

Abstract

The Neurospora VS ribozyme recognizes and cleaves a substrate RNA that contains a GC-rich stem loop. In contrast to most RNA secondary structures that are stable during tertiary or quaternary folding, this substrate undergoes extensive ribozyme-induced rearrangement in the presence of magnesium in which the base pairings of at least seven of the ten nucleotides in the stem are changed. This conformational switch is essential for catalytic activity with the wild-type substrate and creates a metal-binding secondary structure motif near the cleavage site. Base pair rearrangement is accompanied by bulging a cytosine from the middle of the stem, indicating that ribozymes may perform base flipping, an activity previously observed only with protein enzymes that modify DNA.