title: Osmolytes stimulate the reconstitution of functional 50S ribosomes from in vitro transcripts of Escherichia coli 23S rRNA.
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.
authors: Howard Hughes Medical Institute, Department of Molecular Biology and Genetics, Johns Hopkins University School of Medicine, Baltimore, Maryland 21205, USA.
authors: Katharina
authors: K
authors: Semrad
authors: Rachel
authors: R
authors: Green
mesh_terms: D000900:Anti-Bacterial Agents / Q000494:pharmacology; D001483:Base Sequence; D002241:Carbohydrates / Q000494:pharmacology; D002364:Caseins / Q000494:pharmacology; D004926:Escherichia coli / Q000235:genetics*; D048628:Ketolides; D008055:Lipids / Q000494:pharmacology; D018942:Macrolides; D008274:Magnesium / Q000378:metabolism; D008744:Methylamines / Q000494:pharmacology*; D008969:Molecular Sequence Data; D009154:Mutation; D010458:Peptidyl Transferases / Q000187:drug effects / Q000235:genetics / Q000378:metabolism; D014674:Plant Proteins, Dietary / Q000494:pharmacology; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D012338:RNA, Ribosomal, 23S; D012343:RNA, Transfer / Q000235:genetics / Q000378:metabolism; D012270:Ribosomes / Q000187:drug effects / Q000235:genetics* / Q000378:metabolism; D014158:Transcription, Genetic
chemical_list: D000900:Anti-Bacterial Agents; D002241:Carbohydrates; D002364:Caseins; D048628:Ketolides; D008055:Lipids; D018942:Macrolides; D008744:Methylamines; D014674:Plant Proteins, Dietary; D012333:RNA, Messenger; D012338:RNA, Ribosomal, 23S; C080652:Osmolite; D012343:RNA, Transfer; D010458:Peptidyl Transferases; C005855:trimethyloxamine; D008274:Magnesium; C106791:telithromycin