Monitoring of an RNA multistep folding pathway by isothermal titration calorimetry.
Isothermal titration calorimetry was used to monitor the energetic landscape of a catalytic RNA, specifically that of the hepatitis delta virus ribozyme. Using mutants that isolated various tertiary interactions, the thermodynamic parameters of several ribozyme-substrate intermediates were determined. The results shed light on the impact of several tertiary interactions on the global structure of the ribozyme. In addition, the data indicate that the formation of the P1.1 pseudoknot is the limiting step of the molecular mechanism. Last, as illustrated here, isothermal titration calorimetry appears to be a method of choice for the elucidation of an RNA's folding pathway.
Reymond
Cédric
C
RNA Group, Département de Biochimie, Université de Sherbrooke, Sherbrooke, Québec, Canada.
Bisaillon
Martin
M
Perreault
Jean-Pierre
JP
D001345:Autoradiography; D002151:Calorimetry / Q000379:methods; D004591:Electrophoresis, Polyacrylamide Gel; D007700:Kinetics; D009154:Mutation; D009690:Nucleic Acid Conformation; D012313:RNA / Q000737:chemistry* / Q000235:genetics; D016337:RNA, Catalytic / Q000737:chemistry; D013696:Temperature; D013816:Thermodynamics
D016337:RNA, Catalytic; D012313:RNA