In an effort to relate RNA folding to function under conditions more similar to those of a cell, we monitored the self-cleavage reaction of the mammalian CPEB3 human HDV-like ribozyme in the background of physiological salt concentrations, various solutes, and crowding agents. We show that solutes and crowding agents have a stimulatory effect on cleavage at physiological magnesium (0.1 and 0.5 mM Mg2+) while at 10 mM Mg2+ these same additives have virtually no effect on self-cleavage.  We also monitored the effect of solutes and crowding agents on self-cleavage in the background of semi-denaturing conditions of 2.5 M urea and demonstrate that addition of solutes and crowding agents rescue the rate of cleavage to urea-free values in all three Mg2+ concentrations. Lastly SAXS experiments reveal that crowding agents�but not solutes�favor a more compact form of the ribozyme, indicating unique origins of rate enhancement for crowders and solutes.

Cellular-like conditions favor reactivity of a human ribozyme

245th National Meeting (2013)

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