Probing complex RNA structures by mechanical force

Eur Phys J E Soft Matter. 2003 Dec;12(4):605-15. doi: 10.1140/epje/e2004-00033-4.

Abstract

RNA secondary structures of increasing complexity are probed combining single molecule stretching experiments and stochastic unfolding/refolding simulations. We find that force-induced unfolding pathways cannot usually be interpreted by solely invoking successive openings of native helices. Indeed, typical force-extension responses of complex RNA molecules are largely shaped by stretching-induced, long-lived intermediates including non-native helices. This is first shown for a set of generic structural motifs found in larger RNA structures, and then for Escherichia coli's 1540-base long 16S ribosomal RNA, which exhibits a surprisingly well-structured and reproducible unfolding pathway under mechanical stretching. Using out-of-equilibrium stochastic simulations, we demonstrate that these experimental results reflect the slow relaxation of RNA structural rearrangements. Hence, micromanipulations of single RNA molecules probe both their native structures and long-lived intermediates, so-called "kinetic traps", thereby capturing -at the single molecular level- the hallmark of RNA folding/unfolding dynamics.

Publication types

  • Comparative Study
  • Evaluation Study
  • Research Support, Non-U.S. Gov't
  • Validation Study

MeSH terms

  • Computer Simulation
  • Elasticity
  • Escherichia coli / chemistry
  • Micromanipulation / methods*
  • Models, Molecular*
  • Nucleic Acid Conformation*
  • Nucleic Acid Denaturation*
  • Physical Stimulation / methods*
  • RNA / chemistry*
  • RNA / ultrastructure*
  • RNA, Ribosomal, 16S / chemistry
  • Reproducibility of Results
  • Sensitivity and Specificity
  • Stress, Mechanical

Substances

  • RNA, Ribosomal, 16S
  • RNA