Abstract
Functional Escherichia coli 50S ribosomal subunits can be reconstituted from their natural rRNA and protein components. However, when the assembly is performed with in vitro-transcribed 23S rRNA, the reconstitution efficiency is diminished by four orders of magnitude. We tested a variety of chemical chaperones (compounds that are typically used for protein folding), putative RNA chaperones (proteins) and ribosome-targeted antibiotics (small-molecule ligands) that might be reasoned to aid in folding and assembly. Addition of the osmolyte trimethylamine-oxide (TMAO) and the ketolide antibiotic telithromycin (HMR3647) to the reconstitution stimulates its efficiency up to 100-fold yielding a substantially improved system for the in vitro analysis of mutant ribosomes.
Publication types
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Research Support, Non-U.S. Gov't
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Research Support, U.S. Gov't, P.H.S.
MeSH terms
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Anti-Bacterial Agents / pharmacology
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Base Sequence
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Carbohydrates / pharmacology
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Caseins / pharmacology
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Escherichia coli / genetics*
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Ketolides*
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Lipids / pharmacology
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Macrolides*
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Magnesium / metabolism
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Methylamines / pharmacology*
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Molecular Sequence Data
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Mutation
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Peptidyl Transferases / drug effects
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Peptidyl Transferases / genetics
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Peptidyl Transferases / metabolism
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Plant Proteins, Dietary / pharmacology
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RNA, Messenger / genetics
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RNA, Messenger / metabolism
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RNA, Ribosomal, 23S*
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RNA, Transfer / genetics
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RNA, Transfer / metabolism
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Ribosomes / drug effects
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Ribosomes / genetics*
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Ribosomes / metabolism
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Transcription, Genetic
Substances
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Anti-Bacterial Agents
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Carbohydrates
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Caseins
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Ketolides
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Lipids
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Macrolides
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Methylamines
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Plant Proteins, Dietary
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RNA, Messenger
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RNA, Ribosomal, 23S
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Osmolite
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RNA, Transfer
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Peptidyl Transferases
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trimethyloxamine
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Magnesium
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telithromycin