title: Characterization of RNA-protein interactions by phosphorothioate footprinting and its applications to the ribosome.
abstract: Analogs of naturally occurring substances obtained by chemical modifications are powerful tools to study intra- and intermolecular interactions. We have used the phosphorothioate technique to analyze RNA-protein interactions, here the interactions of transfer RNAs (tRNAs) with the three ribosomal binding sites. We describe preparation and purification of thioated tRNAs, formation of functional complexes of programmed ribosomes with tRNAs, and the evaluation of the observed phosphorothioate footprints on the tRNAs.
authors.affiliation: Department of Biochemistry, University of Pretoria, Pretoria, South Africa.
authors.forename: A
authors.initials: A
authors.lastname: Ozlem Tastan Bishop
authors.forename: Ulrich
authors.initials: U
authors.lastname: Stelzl
authors.forename: Markus
authors.initials: M
authors.lastname: Pech
authors.forename: Knud H
authors.initials: KH
authors.lastname: Nierhaus
mesh_terms: D015394:Molecular Structure; D009690:Nucleic Acid Conformation; D010710:Phosphates / Q000737:chemistry*; D011485:Protein Binding; D023302:Protein Footprinting / Q000379:methods*; D011506:Proteins / Q000737:chemistry* / Q000378:metabolism; D012313:RNA / Q000737:chemistry* / Q000378:metabolism; D012270:Ribosomes / Q000737:chemistry* / Q000378:metabolism
chemical_list: D010710:Phosphates; D011506:Proteins; D012313:RNA; C083120:phosphorodithioic acid