The selection in vivo and characterization of an RNA recognition motif for spectinomycin.
Ribonucleoprotein (RNP) complexes participate in almost all macromolecular processes, including RNA processing, protein synthesis, and the signal recognition of proteins targeted for export. An understanding of these processes requires detailed knowledge of interactions at the molecular level, which has evidently been difficult due to the size and complexity of the particles. Fragmentation of large RNP complexes into functional subdomains is proven to be a successful in vitro strategy to probe ligand interactions at the molecular level. We reasoned that RNA molecules expressed in vivo may fold in such a manner as to mimic a drug binding site present on the intact ribosome. If expressed at sufficient levels, the RNA would sequester the antibiotic thereby permitting the continued function of the ribosome and consequently allow the cell to survive in the presence of the drug. Evidence is presented here in support of this RNA fragment-rescue concept following the selection and characterization of RNA fragments that confer resistance to the antibiotic spectinomycin.
Thom
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SmithKline Beecham Pharmaceuticals, Department of Molecular Recognition, Collegeville, PA 19426-0989, USA.
Prescott
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D000900:Anti-Bacterial Agents / Q000378:metabolism* / Q000494:pharmacology; D001483:Base Sequence; D001665:Binding Sites; D004352:Drug Resistance, Microbial; D004926:Escherichia coli / Q000187:drug effects / Q000235:genetics / Q000378:metabolism; D008969:Molecular Sequence Data; D009690:Nucleic Acid Conformation; D012336:RNA, Ribosomal, 16S / Q000378:metabolism*; D012261:Ribonucleoproteins / Q000378:metabolism; D000198:Spectinomycin / Q000378:metabolism* / Q000494:pharmacology; D014169:Transformation, Bacterial
D000900:Anti-Bacterial Agents; D012336:RNA, Ribosomal, 16S; D012261:Ribonucleoproteins; D000198:Spectinomycin