title: Rapid identification and characterization of hammerhead-ribozyme inhibitors using fluorescence-based technology.
abstract: The ability to rapidly identify small molecules that interact with RNA would have significant clinical and research applications. Low-molecular-weight molecules that bind to RNA have the potential to be used as drugs. Therefore, technologies facilitating the rapid and reliable identification of such activities become increasingly important. We have applied a fluorescence-based assay to screen for modulators of hammerhead ribozyme (HHR) catalysis from a small library of antibiotic compounds. Several unknown potent inhibitors of the hammerhead cleavage reaction were identified and further characterized. Tuberactinomycin A, for which positive cooperativity of inhibition in vitro was found, also reduced ribozyme cleavage in vivo. The assay is applicable to the screening of mixtures of compounds, as inhibitory activities were detected within a collection of 2,000 extracts from different actinomycete strains. This approach allows the rapid, reliable, and convenient identification and characterization of ribozyme modulators leading to insights difficult to obtain by classical methodology.
authors.affiliation: Kekulé-Institut für Organische Chemie und Biochemie, Gerhard-Domagk-Str. 1, 35121, Bonn, Germany.
authors.forename: A
authors.initials: A
authors.lastname: Jenne
authors.forename: J S
authors.initials: JS
authors.lastname: Hartig
authors.forename: N
authors.initials: N
authors.lastname: Piganeau
authors.forename: A
authors.initials: A
authors.lastname: Tauer
authors.forename: D A
authors.initials: DA
authors.lastname: Samarsky
authors.forename: M R
authors.initials: MR
authors.lastname: Green
authors.forename: J
authors.initials: J
authors.lastname: Davies
authors.forename: M
authors.initials: M
authors.lastname: Famulok
mesh_terms: D000900:Anti-Bacterial Agents / Q000494:pharmacology*; D001483:Base Sequence; D002384:Catalysis; D015195:Drug Design; D004353:Drug Evaluation, Preclinical / Q000379:methods; D004776:Enviomycin / Q000031:analogs & derivatives* / Q000494:pharmacology; D007700:Kinetics; D009690:Nucleic Acid Conformation; D016337:RNA, Catalytic / Q000037:antagonists & inhibitors / Q000737:chemistry* / Q000378:metabolism*; D013329:Structure-Activity Relationship
chemical_list: D000900:Anti-Bacterial Agents; D016337:RNA, Catalytic; C015563:tuberactinomycin; D004776:Enviomycin