Fidelity of the eukaryotic codon-anticodon interaction: interference by aminoglycoside antibiotics
| Autor: | David C. Eustice, James M. Wilhelm |
|---|---|
| Rok vydání: | 1984 |
| Předmět: |
Biology
Biochemistry Ribosome Structure-Activity Relationship chemistry.chemical_compound Anticodon Animals P-site Codon Binding Sites Aminoglycoside Translation (biology) Hydrogen-Ion Concentration Tetracycline Ribosomal RNA Peptide Elongation Factors Molecular biology Anti-Bacterial Agents A-site chemistry Protein Biosynthesis Tetrahymena Transfer RNA Transfer RNA Aminoacylation Ribosomes Hygromycin B |
| Zdroj: | Biochemistry. 23:1462-1467 |
| ISSN: | 1520-4995 0006-2960 |
| DOI: | 10.1021/bi00302a019 |
| Popis: | A homologous in vitro method was developed from Tetrahymena for ribosomal A-site binding of aminoacyl-tRNA to poly(uridylic acid)-programmed ribosomes with very low error frequency. The reaction mixture pH was the crucial factor in the stable A-site association of aminoacyl-tRNA with high fidelity. At a pH greater than 7.1, endogenous activity translocated A-site-bound aminoacyl-tRNA to the P site. If translocation was allowed to occur, a near-cognate amino-acyl-tRNA, Leu-tRNA, could stably bind to the ribosome by translocation to the ribosomal P site. Near-cognate aminoacyl-tRNA did not stably bind to either site when translocation was blocked. Misreading antibiotics stimulated the stable association of near-cognate aminoacyl-tRNA to the ribosomal A site, thereby increasing the error frequency by several orders of magnitude. Ribosome binding of total aminoacyl-tRNA near equilibrium was not inhibited by misreading antibiotics; however, initial rate kinetics of the binding reaction were dramatically altered such that a 6-fold rate increase was observed with paromomycin or hygromycin B. The rate increase was evident with both cognate and near-cognate aminoacyl-tRNAs. Several antibiotics were tested for misreading potency by the ribosome binding method. We found gentamicin G418 greater than paromomycin greater than neomycin greater than hygromycin B greater than streptomycin in the potentiation of misreading. Tetracycline group antibiotics effectively inhibited A-site aminoacyl-tRNA binding without promoting misreading. |
| Databáze: | OpenAIRE |
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