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Evolution of Tetrahymena ribozyme mutants with increased structural stability
Evolution of Tetrahymena ribozyme mutants with increased structural stability.
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Determining how large RNA molecules stabilize their tertiary structures is critical for understanding how they perform their biological functions. Here we use in vitro selection to identify active variants of the Tetrahymena ribozyme with increased stability. The mutant pool converged to a single family that shared nine mutations; an RNA representing the consensus sequence was structurally more stable by 10.5 degrees C and catalytically active at elevated temperatures. Remarkably, of the nine altered sites, most are already known to be involved in tertiary interactions, and the stabilizing mutations primarily improve the packing interactions in the molecular interior. The wild type ribozyme and the selected mutants provide pairs of mesophilic and thermophilic homologs for studying the origin of their thermal stability.
Guo F, Cech TR
Nature structural biology
2002-11-01 00:00
9
11
855-61
Animals,Directed Molecular Evolution,Evolution, Molecular,Heat,Mutation,Nucleic Acid Conformation,RNA Stability,RNA, Catalytic,RNA, Protozoan,Tetrahymena,RNA, Catalytic,RNA, Protozoan
Howard Hughes Medical Institute, Department of Chemistry and Biochemistry, University of Colorado, Boulder, Colorado 80309-0215, USA
Nat. Struct. Biol.
1072-8368
10.1038/nsb850
nsb850
186
True
12368901
