Key result
Both loop-terminal and middle pairs contribute to the thermodynamic stabilities of 3 x 3 RNA internal loops, with extra stabilization of -1.2 kcal/mol for a GA middle pair flanked by specific pairs.
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Updates RNA thermodynamic parameters for structure prediction; leaves open any clinical translation in RNA therapeutics.
Internal loops in RNA are important for folding and function. The 3 x 3 nucleotide internal loops are the smallest size symmetric loops with a potential noncanonical base pair (middle pair) flanked on both sides by a noncanonical base pair (loop-terminal pair). Thermodynamic and structural insights acquired for 3 x 3 loops should improve approximations for stabilities of 3 x 3 and larger internal loops. Most natural 3 x 3 internal loops are purine rich, which is also true of other internal loops. A series of oligoribonucleotides containing different 3 x 3 internal loops were studied by UV melting and imino proton NMR. Both loop-terminal and middle pairs contribute to the thermodynamic stabilities of 3 x 3 loops. Extra stabilization of -1.2 kcal/mol was found for a GA middle pair when flanked by at least one non-pyrimidine-pyrimidine loop-terminal pair. A penalty of approximately 1 kcal/mol was found for loops with a single loop-terminal GA pair that has a U 3' to the G of the GA pair. A revised model for predicting stabilities of 3 x 3 loops is derived by multiple linear regression.
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Chen et al. (2004) studied this question. Different 3 x 3 internal loops in oligoribonucleotides was evaluated on Thermodynamic stabilities of 3 x 3 loops. Both loop-terminal and middle pairs contribute to the thermodynamic stabilities of 3 x 3 RNA internal loops, with extra stabilization of -1.2 kcal/mol for a GA middle pair flanked by specific pairs.
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