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.
Original language | English (US) |
---|---|
Pages (from-to) | 401-411 |
Number of pages | 11 |
Journal | RNA |
Volume | 8 |
Issue number | 4 |
DOIs | |
State | Published - 2002 |
Keywords
- In vitro reconstitution
- RNA folding
- Ribosome assembly
- Telithromycin
- Trimethylamine oxide (TMAO)
ASJC Scopus subject areas
- Molecular Biology