The self-splicing RNA of Tetrahymena is trapped in a less active conformation by gel purification.
When the circular form of the self-splicing intervening sequence of Tetrahymena thermophila was purified by denaturing polyacrylamide gel electrophoresis by standard methods, the rate of its reaction with tetrauridylate decreased 150-fold at 30 degrees C and at least 1000-fold at 0 degrees C. The activity of the self-splicing RNA was restored by heating it to high temperature and letting it renature in the presence of Mg2+. The rate of reaction of tetrauridylate with the self-splicing RNA flanked by exons was also greatly decreased by gel purification. The difference in activation energies for the reaction of native and denatured intervening sequences suggests that a substantial conformational rearrangement of the gel-purified RNA occurs prior to reaction.
Walstrum
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SA
Department of Chemistry and Biochemistry, University of Colorado, Boulder 80309-0215.
Uhlenbeck
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OC
D000818:Animals; D004591:Electrophoresis, Polyacrylamide Gel; D007700:Kinetics; D008274:Magnesium / Q000494:pharmacology; D009690:Nucleic Acid Conformation; D011487:Protein Conformation; D016337:RNA, Catalytic / Q000302:isolation & purification / Q000378:metabolism*; D013768:Tetrahymena / Q000235:genetics / Q000378:metabolism*; D013816:Thermodynamics
D016337:RNA, Catalytic; D008274:Magnesium