Characterization of RNA-protein interactions by phosphorothioate footprinting and its applications to the ribosome.
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.
Ozlem Tastan Bishop
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Department of Biochemistry, University of Pretoria, Pretoria, South Africa.
Stelzl
Ulrich
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Pech
Markus
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Nierhaus
Knud H
KH
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
D010710:Phosphates; D011506:Proteins; D012313:RNA; C083120:phosphorodithioic acid