Krishna C Suddala1, Nils G Walter2. 1. Biophysics, University of Michigan, Ann Arbor, Michigan, USA; Single Molecule Analysis Group, Department of Chemistry, University of Michigan, Ann Arbor, Michigan, USA. 2. Single Molecule Analysis Group, Department of Chemistry, University of Michigan, Ann Arbor, Michigan, USA. Electronic address: nwalter@umich.edu.
Abstract
Riboswitches are structured noncoding RNA elements that control the expression of their embedding messenger RNAs by sensing the intracellular concentration of diverse metabolites. As the name suggests, riboswitches are dynamic in nature so that studying their inherent conformational dynamics and ligand-mediated folding is important for understanding their mechanism of action. Single-molecule fluorescence energy transfer (smFRET) microscopy is a powerful and versatile technique for studying the folding pathways and intra- and intermolecular dynamics of biological macromolecules, especially RNA. The ability of smFRET to monitor intramolecular distances and their temporal evolution make it a particularly insightful tool for probing the structure and dynamics of riboswitches. Here, we detail the general steps for using prism-based total internal reflection fluorescence microscopy for smFRET studies of the structure, dynamics, and ligand-binding mechanisms of riboswitches.
Riboswitches are structured noncoding RNA elements that control the expression of their embedding messenger RNAs by sensing the intracellular concentration of diverse metabolites. As the name suggests, riboswitches are dynamic in nature so that studying their inherent conformational dynamics and ligan class="Disease">nd-mediated folding is important for understanding their mechanism of action. Single-molecule fluorescence energy transfer (smFRET) microscopy is a powerful and versatile technique for studying the folding pathways and intra- and intermolecular dynamics of biological macromolecules, especially RNA. The ability of smFRET to monitor intramolecular distances and their temporal evolution make it a particularly insightful tool for probing the structure and dynamics of riboswitches. Here, we detail the general steps for using prism-based total internal reflection fluorescence microscopy for smFRET studies of the structure, dynamics, and ligand-binding mechanisms of riboswitches.
Authors: Qinsi Zheng; Manuel F Juette; Steffen Jockusch; Michael R Wasserman; Zhou Zhou; Roger B Altman; Scott C Blanchard Journal: Chem Soc Rev Date: 2014-02-21 Impact factor: 54.564
Authors: Colby D Stoddard; Rebecca K Montange; Scott P Hennelly; Robert P Rambo; Karissa Y Sanbonmatsu; Robert T Batey Journal: Structure Date: 2010-07-14 Impact factor: 5.006
Authors: Michael D Brenner; Mary S Scanlan; Michelle K Nahas; Taekjip Ha; Scott K Silverman Journal: Biochemistry Date: 2010-03-02 Impact factor: 3.162
Authors: Joseph A Liberman; Krishna C Suddala; Asaminew Aytenfisu; Dalen Chan; Ivan A Belashov; Mohammad Salim; David H Mathews; Robert C Spitale; Nils G Walter; Joseph E Wedekind Journal: Proc Natl Acad Sci U S A Date: 2015-06-23 Impact factor: 11.205
Authors: Morgan Chandler; Brittany Johnson; Emil Khisamutdinov; Marina A Dobrovolskaia; Joanna Sztuba-Solinska; Aliasger K Salem; Koen Breyne; Roger Chammas; Nils G Walter; Lydia M Contreras; Peixuan Guo; Kirill A Afonin Journal: ACS Nano Date: 2021-10-22 Impact factor: 18.027