Thiostrepton inhibition of initiation factor 1 activity in polypeptide chain initiation in Escherichia coli.
Thiostrepton, a peptide antibiotic, inhibits the GTP-dependent 70S initiation complex formation (as measured by binding of fMet-tRNA to ribosomes and concomitant hydrolysis of GTP) only when initiation factor 1 is present to permit catalytic recycling of initiation factor 2 in the initiation reaction. When initiation factor 1 is absent, the binding of fMet-tRNA and GTP hydrolysis occur stoichiometrically with respect to initiation factor 2, and thiostrepton has no effect on either reaction under these conditions. Detailed analysis of this inhibition process shows that thiostrepton prevents catalytic recycling of initiation factor 2 by blocking the action of initiation factor 1, which is required for the dissociation of initiation factor 2 from the 70S initiation complex. This dissociation is necessary for the catalytic reutilization of initiation factor 2 in the initiation reaction. The antibiotic does not directly inhibit GTP hydrolysis per se in initiation. The inhibition of fMet-tRNA binding to ribosomes by thiostrepton is also dependent on the concentration of GTP; the inhibition is most pronounced at low concentrations of GTP, but at a high molar ratio of GTP to thiostrepton, the inhibition is completely abolished.
D000900:Anti-Bacterial Agents / Q000494:pharmacology*; D003864:Depression, Chemical; D004926:Escherichia coli / Q000187:drug effects* / Q000378:metabolism; D006160:Guanosine Triphosphate / Q000378:metabolism; D008715:Methionine / Q000378:metabolism; D010442:Peptide Chain Initiation, Translational / Q000187:drug effects*; D010448:Peptide Initiation Factors; D010455:Peptides / Q000494:pharmacology; D010761:Phosphorus Radioisotopes; D011691:Puromycin / Q000378:metabolism; D012343:RNA, Transfer / Q000378:metabolism; D012270:Ribosomes / Q000378:metabolism; D014316:Tritium
D000900:Anti-Bacterial Agents; D010448:Peptide Initiation Factors; D010455:Peptides; D010761:Phosphorus Radioisotopes; D014316:Tritium; D011691:Puromycin; D006160:Guanosine Triphosphate; D012343:RNA, Transfer; D008715:Methionine