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.
Department of Chemistry, University of Utah, Salt Lake City 84112, USA.
Momchilo
M
Vuyisich
Peter A
PA
Beal
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