An RNA motif that binds ATP

Area of Science:

• Molecular Biology
• Biochemistry
• RNA Therapeutics

Background:

• RNA molecules can be engineered to bind specific small molecule ligands.
• Isolating high-affinity RNA binders from large random pools is challenging.
• Existing methods often rely on solid supports, limiting solution-phase binding discovery.

Purpose of the Study:

• To develop an improved in vitro selection method for isolating RNA ligands that bind targets in solution.
• To identify and characterize novel RNA motifs with high-affinity binding to Adenosine Triphosphate (ATP).

Main Methods:

• A novel in vitro selection strategy was employed to isolate RNA molecules binding to a target ligand.
• The selection process was optimized to ensure isolation of RNAs binding in solution.
• Characterization of RNA-ligand interactions involved binding assays with analogues and modified RNAs.

Main Results:

• A small, consensus RNA motif with high-affinity for ATP was successfully isolated.
• The selected RNA motif exhibits a common secondary structure essential for binding.
• Detailed analysis revealed extensive contacts and conformational changes upon ATP binding.

Conclusions:

• The new selection procedure effectively isolates functional RNA ligands from complex mixtures.
• The identified ATP-binding RNA motif offers insights into molecular recognition mechanisms.
• This work provides a foundation for designing RNA-based sensors and therapeutics.