Dynamics-Function Analysis in Catalytic RNA Using NMR Spin Relaxation and Conformationally Restricted Nucleotides

Area of Science:

• Biochemistry
• Structural Biology
• Molecular Biophysics

Background:

• Understanding biomolecular function necessitates analyzing dynamic properties and their role in function.
• RNA's dynamic nature is critical for its diverse biological roles.
• Existing methods may not fully capture the functional relevance of specific RNA dynamics.

Purpose of the Study:

• To develop and apply NMR methods for identifying molecular disorder and conformational transitions in RNA.
• To assess the functional importance of specific RNA dynamics by manipulating structural ensembles.
• To link RNA backbone dynamics to overall biomolecular function.

Main Methods:

• Utilizing Nuclear Magnetic Resonance (NMR) spectroscopy.
• Employing metabolically directed specific isotope labeling for RNA analysis.
• Incorporating synthetic covalently modified nucleotides with constrained sugar puckers.

Main Results:

• Identification of molecular disorder and/or conformational transitions on RNA backbone ribose groups.
• Assessment of dynamics by selectively removing minor conformers identified via NMR.
• Demonstration of a method to functionally evaluate specific dynamic states within RNA structures.

Conclusions:

• NMR with specific isotope labeling is effective for characterizing RNA backbone dynamics.
• Synthetic nucleotide analogs are valuable tools for probing the functional significance of RNA conformational heterogeneity.
• This integrated approach enhances the understanding of structure-dynamics-function relationships in RNA.