Abstract
Helix 89 of the 23S rRNA connects ribosomal peptidyltransferase center and elongation factor binding site. Secondary structure of helix 89 determined by X-ray structural analysis involves less base pairs then could be drawn for the helix of the same primary structure. It can be that alternative secondary structure might be realized at some stage of translation. Here by means of site-directed mutagenesis we stabilized either the "X-ray" structure or the structure with largest number of paired nucleotides. Mutation UU2492-3C which aimed to provide maximal pairing of the helix 89 of the 23S rRNA was lethal. Mutant ribosomes were unable to catalyze peptide transfer independently either with aminoacyl-tRNA or puromycin.
Copyright © 2011. Published by Elsevier B.V.
Publication types
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Research Support, Non-U.S. Gov't
MeSH terms
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Base Sequence
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Crystallography, X-Ray
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Dipeptides / biosynthesis
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Escherichia coli / genetics*
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Escherichia coli / metabolism*
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Molecular Sequence Data
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Mutagenesis, Site-Directed
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Nucleic Acid Conformation*
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Peptidyl Transferases / genetics
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Peptidyl Transferases / metabolism*
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Protein Biosynthesis
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Protein Structure, Secondary
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Protein Synthesis Inhibitors / metabolism
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Puromycin / metabolism
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RNA, Ribosomal, 23S / chemistry*
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RNA, Ribosomal, 23S / genetics
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RNA, Ribosomal, 23S / metabolism
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RNA, Transfer, Amino Acyl / chemistry
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RNA, Transfer, Amino Acyl / genetics
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RNA, Transfer, Amino Acyl / metabolism
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Ribosomes / genetics
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Ribosomes / metabolism*
Substances
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Dipeptides
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Protein Synthesis Inhibitors
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RNA, Ribosomal, 23S
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RNA, Transfer, Amino Acyl
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Puromycin
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Peptidyl Transferases