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Codon-anticodon interaction at the P site is a prerequisite for tRNA interaction with the small ribosomal subunit.
Schäfer, Markus A; Tastan, A Ozlem; Patzke, Sebastian; Blaha, Gregor; Spahn, Christian M T; Wilson, Daniel N; Nierhaus, Knud H.
Afiliación
  • Schäfer MA; Max-Planck-Institut für Molekulare Genetik, AG Ribosomen, Ihnestrasse 73, D-14195 Berlin, Germany.
J Biol Chem ; 277(21): 19095-105, 2002 May 24.
Article en En | MEDLINE | ID: mdl-11867615
The arrival of high resolution crystal structures for the ribosomal subunits opens a new phase of molecular analysis and asks for corresponding analyses of ribosomal function. Here we apply the phosphorothioate technique to dissect tRNA interactions with the ribosome. We demonstrate that a tRNA bound to the P site of non-programmed 70 S ribosomes contacts predominantly the 50 S, as opposed to the 30 S subunit, indicating that codon-anticodon interaction at the P site is a prerequisite for 30 S binding. Protection patterns of tRNAs bound to isolated subunits and programmed 70 S ribosomes were compared. The results suggest the presence of a movable domain in the large ribosomal subunit that carries tRNA and reveal that only approximately 15% of a tRNA, namely residues 30 +/- 1 to 43 +/- 1, contact the 30 S subunit of programmed 70 S ribosomes, whereas the remaining 85% make contact with the 50 S subunit. Identical protection patterns of two distinct elongator tRNAs at the P site were identified as tRNA species-independent phosphate backbone contacts. The sites of protection correlate nicely with the predicted ribosomal-tRNA contacts deduced from a 5.5-A crystal structure of a programmed 70 S ribosome, thus refining which ribosomal components are critical for tRNA fixation at the P site.
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Colección: 01-internacional Base de datos: MEDLINE Asunto principal: Ribosomas / Anticodón / Codón / ARN de Transferencia Tipo de estudio: Prognostic_studies Idioma: En Revista: J Biol Chem Año: 2002 Tipo del documento: Article País de afiliación: Alemania Pais de publicación: Estados Unidos
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Colección: 01-internacional Base de datos: MEDLINE Asunto principal: Ribosomas / Anticodón / Codón / ARN de Transferencia Tipo de estudio: Prognostic_studies Idioma: En Revista: J Biol Chem Año: 2002 Tipo del documento: Article País de afiliación: Alemania Pais de publicación: Estados Unidos