INSTITUTO DE QUIMICA, FISICA DE LOS MATERIALES, MEDIOAMBIENTE Y ENERGIA
Unidad Ejecutora - UE
The Role of Counterion Valence and Size in GAAA 6 TetraloopReceptor Docking/Undocking Kinetics
JULIE L. FIORE; ERIK D. HOLMSTROM; LARRY R. FIEGLAND; JOSE H. HODAK; DAVID J. NESBITT
JOURNAL OF MOLECULAR BIOLOGY
ACADEMIC PRESS LTD-ELSEVIER SCIENCE LTD
Lugar: Amsterdam; Año: 2012
For RNA to fold into compact, ordered structures, it must overcome electrostatic repulsion between negatively charged phosphate groups by counterion recruitment.Aphysical understanding of the counterion-assisted folding process requires addressing how cations kinetically and thermodynamically control the folding equilibrium for each tertiary interaction in a full‐length RNA. In this work, single-molecule FRET (fluorescence resonance energy transfer) techniques are exploited to isolate and explore the cation concentration‐dependent kinetics for formation of a ubiquitousRNAtertiary interaction, that is, the docking/undocking of a GAAA tetraloop with its 11‐nt receptor. Rate constants for docking (kdock) and undocking (kundock) are obtained as a function of cation concentration, size, and valence, specifically for the series Na+, K+, Mg2+, Ca2+, Co(NH3)6+, and spermidine3+. Increasing cation concentration accelerates kdock dramatically but achieves only a slight decrease in kundock. These results can be kinetically modeled using parallel cation-dependent and cation‐independent docking pathways, which allows for isolation of the folding kinetics from the interaction energetics of the cations with the undocked and docked states, respectively. This analysis reveals a preferential interaction of the cations with the transition state and docked state as compared to the undocked RNA, with the ionRNA interaction strength growing with cation valence. However, the corresponding number of cations that are taken up by the RNA upon folding decreases with charge density of the cation. The only exception to these behaviors is spermidine3+, whose weaker influence on the docking equilibria with respect to Co(NH3)6 3+ can be ascribed to steric effects preventing completeneutralization of the RNA phosphate groups.