PMID:11991636

Citation	Semrad, K and Green, R (2002) Osmolytes stimulate the reconstitution of functional 50S ribosomes from in vitro transcripts of Escherichia coli 23S rRNA. RNA 8:401-11
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.
Links	PubMed PMC1370264
Keywords	Anti-Bacterial Agents/pharmacology; Base Sequence; Carbohydrates/pharmacology; Caseins/pharmacology; Escherichia coli/genetics; Ketolides; Lipids/pharmacology; Macrolides; Magnesium/metabolism; Methylamines/pharmacology; Molecular Sequence Data; Mutation; Peptidyl Transferases/drug effects; Peptidyl Transferases/genetics; Peptidyl Transferases/metabolism; RNA, Messenger/genetics; RNA, Messenger/metabolism; RNA, Ribosomal, 23S; RNA, Transfer/genetics; RNA, Transfer/metabolism; Ribosomes/drug effects; Ribosomes/genetics; Ribosomes/metabolism; Transcription, Genetic; Vegetable Proteins/pharmacology
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