In vivo recognition of an RNA aptamer by its transcription factor target.
In vitro-selected RNA aptamers are potential inhibitors of disease-related macromolecules. Our laboratory previously isolated an RNA aptamer that specifically binds to the human transcription factor NF-kappaB. This RNA aptamer competitively inhibits DNA binding by NF-kappaB in vitro. In the study presented here, this aptamer was tested for binding to the p50 homodimer form of NF-kappaB (p50(2)) in eukaryotic cells using a yeast three-hybrid system. We show that the alpha-p50 RNA aptamer selectively binds recombinant p50(2) expressed in yeast, demonstrating in vivo recognition of an in vitro-selected RNA aptamer by its protein target. This result suggests that RNA decoys might be used to inhibit the function of DNA-binding proteins in vivo.
Cassiday
L A
LA
Department of Biochemistry and Molecular Biology, Mayo Foundation, Rochester, Minnesota 55905, USA.
Maher
L J
LJ
D000818:Animals; D001483:Base Sequence; D001667:Binding, Competitive / Q000235:genetics; D019281:Dimerization; D005822:Genetic Vectors / Q000138:chemical synthesis / Q000378:metabolism; D006801:Humans; D051379:Mice; D008969:Molecular Sequence Data; D016328:NF-kappa B / Q000235:genetics / Q000378:metabolism* / Q000502:physiology; D052002:NF-kappa B p50 Subunit; D009690:Nucleic Acid Conformation; D012331:RNA, Fungal / Q000235:genetics / Q000378:metabolism* / Q000502:physiology; D011817:Rabbits; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis / Q000138:chemical synthesis / Q000378:metabolism; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism; D015203:Reproducibility of Results; D012441:Saccharomyces cerevisiae / Q000235:genetics; D020798:Two-Hybrid System Techniques
D016328:NF-kappa B; D052002:NF-kappa B p50 Subunit; D012331:RNA, Fungal; D011993:Recombinant Fusion Proteins; D011994:Recombinant Proteins