Thiophosphate interference experiments locate phosphates important for the hammerhead RNA self-cleavage reaction.
A hammerhead domain of less than 50 nucleotides is responsible for a self-cleavage reaction in the replication of plant RNA pathogens. The hammerhead is composed of three helices joining at a central conserved core of 11 single stranded nucleotides. The core is believed to fold into a tertiary structure that provides functional groups for catalysis and to coordinate one or more divalent metal ions. In this study we use a phosphorothioate substitution interference assay to identify four phosphates in the conserved core which also play a role in the self-cleavage reaction.
Ruffner
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Department of Chemistry and Biochemistry, University of Colorado, Boulder 80309.
Uhlenbeck
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D012321:DNA-Directed RNA Polymerases / Q000378:metabolism; D004926:Escherichia coli / Q000201:enzymology; D007700:Kinetics; D009690:Nucleic Acid Conformation; D012326:RNA Splicing / Q000187:drug effects*; D016337:RNA, Catalytic / Q000378:metabolism*; D012265:Ribonucleotides / Q000494:pharmacology*; D013329:Structure-Activity Relationship; D013604:T-Phages / Q000201:enzymology; D013873:Thionucleotides / Q000494:pharmacology*; D014158:Transcription, Genetic / Q000187:drug effects*
D016337:RNA, Catalytic; D012265:Ribonucleotides; D013873:Thionucleotides; D012321:DNA-Directed RNA Polymerases