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.
Jaikaran
D C
DC
Department of Chemistry, University of Toronto, Ontario, Canada.
MacMillan
A M
AM
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