Template-directed interference footprinting for RNA based on inosine-specific cleavage.
We report here the development of a Template-directed Interference (TDI) footprinting assay for RNA. The TDI nucleotide analogue inosine (I) lacks the exocyclic amine of G and is a suitable probe for the role of this group in RNA structure and function. Using an I-specific cleavage protocol we identified three functionally significant G residues in the Tetrahymena ribozyme. These residues are proximal to the active site of the folded intron and likely contribute to the positioning of substrates at the catalytic core.
Department of Chemistry, University of Toronto, Ontario, Canada.
D C
DC
Jaikaran
A M
AM
MacMillan
D000818:Animals; D018983:DNA Footprinting; D006868:Hydrolysis; D007288:Inosine / Q000737:chemistry*; D023282:Nuclease Protection Assays; D009690:Nucleic Acid Conformation; D012313:RNA / Q000737:chemistry*; D013698:Templates, Genetic; D013768:Tetrahymena / Q000235:genetics
D007288:Inosine; D012313:RNA