Evolution of Tetrahymena ribozyme mutants with increased structural stability.
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.
D000818:Animals; D019020:Directed Molecular Evolution; D019143:Evolution, Molecular; D006358:Hot Temperature; D009154:Mutation; D009690:Nucleic Acid Conformation; D020871:RNA Stability; D016337:RNA, Catalytic / Q000737:chemistry / Q000235:genetics*; D016053:RNA, Protozoan / Q000737:chemistry / Q000235:genetics*; D013768:Tetrahymena / Q000235:genetics*
D016337:RNA, Catalytic; D016053:RNA, Protozoan