Controlling protein activity with ligand-regulated RNA aptamers.
Controlling the activity of a protein is necessary for defining its function in vivo. RNA aptamers are capable of inhibiting proteins with high affinity and specificity, but this effect is not readily reversible. We describe a general method for discovering aptamers that bind and inhibit their target protein, but addition of a specific small molecule disrupts the protein-RNA complex. A SELEX protocol was used to raise RNA aptamers to the DNA repair enzyme, formamidopyrimidine glycosylase (Fpg), and neomycin was employed in each round to dissociate Fpg-bound RNAs. We identified an RNA molecule able to completely inhibit Fpg at 100 nM concentration. Importantly, Fpg activity is recovered by the addition of neomycin. We envision these ligand-regulated aptamers (LIRAs) as valuable tools in the study of biological phenomena in which the timing of molecular events is critical.
Vuyisich
Momchilo
M
Department of Chemistry, University of Utah, Salt Lake City 84112, USA.
Beal
Peter A
PA
D001426:Bacterial Proteins / Q000037:antagonists & inhibitors*; D001665:Binding Sites; D020650:Combinatorial Chemistry Techniques; D043382:DNA-Formamidopyrimidine Glycosylase; D004336:Drug Antagonism; D008024:Ligands; D009699:N-Glycosyl Hydrolases / Q000037:antagonists & inhibitors*; D009355:Neomycin / Q000494:pharmacology; D009690:Nucleic Acid Conformation / Q000187:drug effects; D011485:Protein Binding / Q000187:drug effects; D023302:Protein Footprinting; D012313:RNA / Q000737:chemistry / Q000187:drug effects / Q000494:pharmacology*
D001426:Bacterial Proteins; D008024:Ligands; D012313:RNA; D009699:N-Glycosyl Hydrolases; D043382:DNA-Formamidopyrimidine Glycosylase; D009355:Neomycin