Single synonymous codon substitution eliminates pausing during chloramphenicol acetyl transferase synthesis onEscherichia coliribosomes in vitro
| Autor: | Paul M. Horowitz, Boyd Hardesty, Gisela Kramer, Vasanthi Ramachandiran |
|---|---|
| Rok vydání: | 2002 |
| Předmět: |
Chloramphenicol O-Acetyltransferase
Nascent polypeptide Molecular Sequence Data Peptide Chain Elongation Translational Biophysics Biology medicine.disease_cause Biochemistry Ribosome Rare codon Translational pausing Structural Biology Escherichia coli Genetics medicine Coding region Isoleucine Codon Molecular Biology Peptide sequence Messenger RNA Base Sequence Cell-Free System In vitro protein synthesis Cell Biology Molecular biology Mutagenesis Codon usage bias Transfer RNA Nucleic Acid Conformation Ribosomes |
| Zdroj: | FEBS Letters. 512:209-212 |
| ISSN: | 0014-5793 |
| Popis: | The coding sequence for chloramphenicol acetyl transferase (CAT) contains several rare codons; three of them are ATA encoding isoleucine in positions 13, 84 and 119 of the amino acid sequence. Expression of CAT on Escherichia coli ribosomes in vitro results in mostly full-length product but also distinct smaller polypeptides from less than 3 kDa to over 20 kDa. As reported earlier, the smaller polypeptides are the predominant products, if translation is initiated with fluorophore-Met-tRNAf. All this translational pausing is eliminated when the first ATA codon is mutated to ATC, a frequently used codon for isoleucine in E. coli. Addition of large amounts of E. coli tRNA to the coupled transcription/translation reaction does not reduce the number of pause-site peptides seen in the expression of wild-type CAT. Thus we hypothesize that the mRNA structure may be an important determinant for translational pausing. |
| Databáze: | OpenAIRE |
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