title: Translation elongation by a hybrid ribosome in which proteins at the GTPase center of the Escherichia coli ribosome are replaced with rat counterparts.
abstract: Ribosomal L10-L7/L12 protein complex and L11 bind to a highly conserved RNA region around position 1070 in domain II of 23 S rRNA and constitute a part of the GTPase-associated center in Escherichia coli ribosomes. We replaced these ribosomal proteins in vitro with the rat counterparts P0-P1/P2 complex and RL12, and tested them for ribosomal activities. The core 50 S subunit lacking the proteins on the 1070 RNA domain was prepared under gentle conditions from a mutant deficient in ribosomal protein L11. The rat proteins bound to the core 50 S subunit through their interactions with the 1070 RNA domain. The resultant hybrid ribosome was insensitive to thiostrepton and showed poly(U)-programmed polyphenylalanine synthesis dependent on the actions of both eukaryotic elongation factors 1alpha (eEF-1alpha) and 2 (eEF-2) but not of the prokaryotic equivalent factors EF-Tu and EF-G. The results from replacement of either the L10-L7/L12 complex or L11 with rat protein showed that the P0-P1/P2 complex, and not RL12, was responsible for the specificity of the eukaryotic ribosomes to eukaryotic elongation factors and for the accompanying GTPase activity. The presence of either E. coli L11 or rat RL12 considerably stimulated the polyphenylalanine synthesis by the hybrid ribosome, suggesting that L11/RL12 proteins play an important role in post-GTPase events of translation elongation.
authors.affiliation: Institute of High Polymer Research, Faculty of Textile Science and Technology, Shinshu University, Ueda 386-8567, Japan. uchiumi@giptc.shinshu-u.ac.jp
authors.forename: Toshio
authors.initials: T
authors.lastname: Uchiumi
authors.forename: Sachiko
authors.initials: S
authors.lastname: Honma
authors.forename: Takaomi
authors.initials: T
authors.lastname: Nomura
authors.forename: Eric R
authors.initials: ER
authors.lastname: Dabbs
authors.forename: Akira
authors.initials: A
authors.lastname: Hachimori
mesh_terms: D000818:Animals; D004926:Escherichia coli / Q000378:metabolism*; D020558:GTP Phosphohydrolases / Q000378:metabolism*; D010441:Peptide Chain Elongation, Translational / Q000502:physiology*; D051381:Rats; D012270:Ribosomes / Q000502:physiology*
chemical_list: D020558:GTP Phosphohydrolases