title: Enthalpy-driven RNA folding: single-molecule thermodynamics of tetraloop-receptor tertiary interaction.
abstract: RNA folding thermodynamics are crucial for structure prediction, which requires characterization of both enthalpic and entropic contributions of tertiary motifs to conformational stability. We explore the temperature dependence of RNA folding due to the ubiquitous GAAA tetraloop-receptor docking interaction, exploiting immobilized and freely diffusing single-molecule fluorescence resonance energy transfer (smFRET) methods. The equilibrium constant for intramolecular docking is obtained as a function of temperature (T = 21-47 degrees C), from which a van't Hoff analysis yields the enthalpy (DeltaH degrees) and entropy (DeltaS degrees) of docking. Tetraloop-receptor docking is significantly exothermic and entropically unfavorable in 1 mM MgCl(2) and 100 mM NaCl, with excellent agreement between immobilized (DeltaH degrees = -17.4 +/- 1.6 kcal/mol, and DeltaS degrees = -56.2 +/- 5.4 cal mol(-1) K(-1)) and freely diffusing (DeltaH degrees = -17.2 +/- 1.6 kcal/mol, and DeltaS degrees = -55.9 +/- 5.2 cal mol(-1) K(-1)) species. Kinetic heterogeneity in the tetraloop-receptor construct is unaffected over the temperature range investigated, indicating a large energy barrier for interconversion between the actively docking and nondocking subpopulations. Formation of the tetraloop-receptor interaction can account for approximately 60% of the DeltaH degrees and DeltaS degrees of P4-P6 domain folding in the Tetrahymena ribozyme, suggesting that it may act as a thermodynamic clamp for the domain. Comparison of the isolated tetraloop-receptor and other tertiary folding thermodynamics supports a theme that enthalpy- versus entropy-driven folding is determined by the number of hydrogen bonding and base stacking interactions.
authors.affiliation: JILA, National Institute of Standards and Technology and University of Colorado, Boulder, Colorado 80309-0440, USA.
authors.forename: Julie L
authors.initials: JL
authors.lastname: Fiore
authors.forename: Benedikt
authors.initials: B
authors.lastname: Kraemer
authors.forename: Felix
authors.initials: F
authors.lastname: Koberling
authors.forename: Rainer
authors.initials: R
authors.lastname: Edmann
authors.forename: David J
authors.initials: DJ
authors.lastname: Nesbitt
mesh_terms: D007700:Kinetics; D009690:Nucleic Acid Conformation; D011485:Protein Binding; D011506:Proteins / Q000737:chemistry*; D012313:RNA / Q000737:chemistry*; D016337:RNA, Catalytic / Q000737:chemistry; D013696:Temperature; D013768:Tetrahymena / Q000737:chemistry; D013816:Thermodynamics
chemical_list: D011506:Proteins; D016337:RNA, Catalytic; D012313:RNA