Cooperative Analysis of Structural Dynamics in RNA-Protein Complexes by Single-Molecule Förster Resonance Energy Transfer Spectroscopy

Molecules. 2020 Apr 28;25(9):2057. doi: 10.3390/molecules25092057.

Abstract

RNA-protein complexes (RNPs) are essential components in a variety of cellular processes, and oftentimes exhibit complex structures and show mechanisms that are highly dynamic in conformation and structure. However, biochemical and structural biology approaches are mostly not able to fully elucidate the structurally and especially conformationally dynamic and heterogeneous nature of these RNPs, to which end single molecule Förster resonance energy transfer (smFRET) spectroscopy can be harnessed to fill this gap. Here we summarize the advantages of strategic smFRET studies to investigate RNP dynamics, complemented by structural and biochemical data. Focusing on recent smFRET studies of three essential biological systems, we demonstrate that investigation of RNPs on a single molecule level can answer important functional questions that remained elusive with structural or biochemical approaches alone: The complex structural rearrangements throughout the splicing cycle, unwinding dynamics of the G-quadruplex (G4) helicase RHAU, and aspects in telomere maintenance regulation and synthesis.

Keywords: G quadruplex helicase RHAU; RNA-protein complex (RNP); RNP dynamics; single-molecule förster resonance energy transfer (smFRET) spectroscopy; spliceosome; telomerase.

Publication types

  • Review

MeSH terms

  • Animals
  • Cattle
  • Cluster Analysis
  • Crystallography, X-Ray
  • Fluorescence Resonance Energy Transfer*
  • G-Quadruplexes*
  • Humans
  • Markov Chains
  • Nucleic Acid Conformation
  • Protein Binding
  • Protein Denaturation
  • Protein Folding
  • Protein Structure, Secondary
  • RNA / chemistry*
  • RNA Splicing
  • Ribonucleoproteins
  • Single Molecule Imaging*
  • Spliceosomes / chemistry
  • Telomerase / chemistry
  • Telomere / chemistry
  • Telomere / ultrastructure

Substances

  • Ribonucleoproteins
  • RNA
  • Telomerase