Concerted kinetic folding of a multidomain ribozyme with a disrupted loop-receptor interaction

J Mol Biol. 2001 Jan 5;305(1):11-21. doi: 10.1006/jmbi.2000.4253.

Abstract

The free energy landscape for the folding of large, multidomain RNAs is rugged, and kinetically trapped, misfolded intermediates are a hallmark of RNA folding reactions. Here, we examine the role of a native loop-receptor interaction in determining the ruggedness of the energy landscape for folding of the Tetrahymena ribozyme. We demonstrate a progressive smoothing of the energy landscape for ribozyme folding as the strength of the loop-receptor interaction is reduced. Remarkably, with the most severe mutation, global folding is more rapid than for the wild-type ribozyme and proceeds in a concerted fashion without the accumulation of long-lived kinetic intermediates. The results demonstrate that a complex interplay between native tertiary interactions, divalent ion concentration, and non-native secondary structure determines the ruggedness of the energy landscape. Furthermore, the results suggest that kinetic folding transitions involving large regions of highly structured RNAs can proceed in a concerted fashion, in the absence of significant stable, preorganized tertiary structure.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Animals
  • Base Pairing / genetics
  • Base Sequence
  • Binding Sites
  • Cations, Divalent / metabolism
  • Kinetics
  • Magnesium / metabolism
  • Models, Chemical
  • Mutation / genetics
  • Nucleic Acid Conformation*
  • Nucleic Acid Hybridization
  • Oligoribonucleotides / chemistry
  • Oligoribonucleotides / genetics
  • Oligoribonucleotides / metabolism
  • RNA Stability / genetics*
  • RNA, Catalytic / chemistry*
  • RNA, Catalytic / genetics
  • RNA, Catalytic / metabolism*
  • Tetrahymena / genetics*
  • Thermodynamics

Substances

  • Cations, Divalent
  • Oligoribonucleotides
  • RNA, Catalytic
  • Magnesium