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Metal ion induced heterogeneity in RNA folding studied by smFRET

Type of publication Peer-reviewed
Publikationsform Original article (peer-reviewed)
Author Börner Richard, Kowerko Danny, Miserachs Helena Guiset, Schaffer Michelle F., Sigel Roland K.O.,
Project Metal Ions in Structure and Function of Regulatory RNAs
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Original article (peer-reviewed)

Journal Coordination Chemistry Reviews
Volume (Issue) 327-328
Page(s) 123 - 142
Title of proceedings Coordination Chemistry Reviews
DOI 10.1016/j.ccr.2016.06.002


More than two decades of investigating nucleic acids and ribonucleic acids (RNA) using single molecule Förster resonance energy transfer (smFRET) have passed. It turned out that sample heterogeneity in structure and function of RNA molecules as well as folding intermediates, kinetic subpopulations, and interconversion rates of conformational states of RNA biomolecules, all of which are usually hidden in ensemble type experiments, are often observed characteristics. Besides proteins, metal ions play a crucial role in RNA folding and dynamics, as well as RNA/RNA or RNA/DNA interactions. RNA molecules form discrete conformational intermediates before reaching the native three-dimensional fold, whereby metal ions guide the folding pathway by changing the energetic barriers between local and global minima in the energy landscape. Here we review recent advances in the characterization of the role of metal ions in folding and function of nucleic acid structures by means of smFRET. Subsequently, the workflow of smFRET data analysis is described and exemplified by the metal ion-depending folding and dynamics of the group IIB intron from Saccharomyces cerevisiae and RNA–RNA binding kinetics of this ribozyme's 5'-splice site formation.