Folding mechanisms, therefore, play significant roles in RNA function and related biological phenomena. RNA molecules fold into unique three dimensional structures, which allow them to function as catalysts that perform biochemical reactions and regulators that control protein expression 1. Overall, these results indicate that the states of Mg 2+ should not be limited to free and bound only, as weakly bound Mg 2+ strongly promotes RNA function under cellular conditions. Other metabolites that weakly chelate magnesium offer similar beneficial effects, which implies chelated magnesium may enhance RNA function in the cell in the same way. Furthermore, aaCM enhance self-cleavage of several different ribozymes, up to 100,000-fold at Mg 2+ free of just 0.5 mM, indirectly through RNA compaction. Amino acid-chelated Mg 2+ (aaCM) of ~15 mM dramatically increases RNA folding and prevents RNA degradation. To uncover cellular behavior of RNA, we devised cytoplasm mimic systems that include biological concentrations of amino acids, which weakly chelate Mg 2+. Most RNA folding studies have been performed under non-physiological conditions of high concentrations (≥10 mM) of Mg 2+ free, while actual cellular concentrations of Mg 2+ free are only ~1 mM in a background of greater than 50 mM Mg 2+ total.
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