FRET-guided modeling of nucleic acids.
Nucleic Acids Res
; 52(13): e59, 2024 Jul 22.
Article
in En
| MEDLINE
| ID: mdl-38869063
ABSTRACT
The functional diversity of RNAs is encoded in their innate conformational heterogeneity. The combination of single-molecule spectroscopy and computational modeling offers new attractive opportunities to map structural transitions within nucleic acid ensembles. Here, we describe a framework to harmonize single-molecule Förster resonance energy transfer (FRET) measurements with molecular dynamics simulations and de novo structure prediction. Using either all-atom or implicit fluorophore modeling, we recreate FRET experiments in silico, visualize the underlying structural dynamics and quantify the reaction coordinates. Using multiple accessible-contact volumes as a post hoc scoring method for fragment assembly in Rosetta, we demonstrate that FRET can be used to filter a de novo RNA structure prediction ensemble by refuting models that are not compatible with in vitro FRET measurement. We benchmark our FRET-assisted modeling approach on double-labeled DNA strands and validate it against an intrinsically dynamic manganese(II)-binding riboswitch. We show that a FRET coordinate describing the assembly of a four-way junction allows our pipeline to recapitulate the global fold of the riboswitch displayed by the crystal structure. We conclude that computational fluorescence spectroscopy facilitates the interpretability of dynamic structural ensembles and improves the mechanistic understanding of nucleic acid interactions.
Full text:
1
Collection:
01-internacional
Database:
MEDLINE
Main subject:
Fluorescence Resonance Energy Transfer
/
Molecular Dynamics Simulation
/
Riboswitch
/
Nucleic Acid Conformation
Language:
En
Journal:
Nucleic Acids Res
Year:
2024
Type:
Article
Affiliation country:
Switzerland