Combining Coarse-Grained Simulations and Single Molecule Analysis Reveals a Three-State Folding Model of the Guanidine-II Riboswitch

Author(s)
Christin Fuks, Sebastian Falkner, Nadine Schwierz, Martin Hengesbach
Abstract

Riboswitch RNAs regulate gene expression by conformational changes induced by environmental conditions and specific ligand binding. The guanidine-II riboswitch is proposed to bind the small molecule guanidinium and to subsequently form a kissing loop interaction between the P1 and P2 hairpins. While an interaction was shown for isolated hairpins in crystallization and electron paramagnetic resonance experiments, an intrastrand kissing loop formation has not been demonstrated. Here, we report the first evidence of this interaction in cis in a ligand and Mg

2+ dependent manner. Using single-molecule FRET spectroscopy and detailed structural information from coarse-grained simulations, we observe and characterize three interconvertible states representing an open and kissing loop conformation as well as a novel Mg

2+ dependent state for the guanidine-II riboswitch from E. coli. The results further substantiate the proposed switching mechanism and provide detailed insight into the regulation mechanism for the guanidine-II riboswitch class. Combining single molecule experiments and coarse-grained simulations therefore provides a promising perspective in resolving the conformational changes induced by environmental conditions and to yield molecular insights into RNA regulation.

Organisation(s)
Computational and Soft Matter Physics
External organisation(s)
Johann Wolfgang Goethe-Universität Frankfurt am Main, Max Planck Institute of Biophysics
Journal
Frontiers in Molecular Biosciences
Volume
9
No. of pages
16
ISSN
2296-889X
DOI
https://doi.org/10.3389/fmolb.2022.826505
Publication date
04-2022
Peer reviewed
Yes
Austrian Fields of Science 2012
106041 Structural biology, 103029 Statistical physics
Keywords
ASJC Scopus subject areas
Biochemistry, Genetics and Molecular Biology (miscellaneous), Molecular Biology, Biochemistry
Portal url
https://ucrisportal.univie.ac.at/en/publications/be5eb37a-1ac5-448e-9f77-d0996257fe24