Crucial contribution of the multiple copies of the initiator tRNA genes in the fidelity of tRNAfMet selection on the ribosomal P-site in Escherichia coli

摘要

The accuracy of the initiator tRNA (tRNA^fMet) selection in the ribosomal P-site is central to the fidelity of protein synthesis. A highly conserved occurrence of three consecutive G–C base pairs in the anticodon stem of tRNA^fMet contributes to its preferential selection in the P-site. In a genetic screen, using a plasmid borne copy of an inactive tRNA^fMet mutant wherein the three G–C base pairs were changed, we isolated Escherichia coli strains that allow efficient initiation with the tRNA^fMet mutant. Here, extensive characterization of two such strains revealed novel mutations in the metZWV promoter severely compromising tRNA^fMet levels. Low cellular abundance of the chromosomally encoded tRNA^fMet allows efficient initiation with the tRNA^fMet mutant and an elongator tRNA^Gln, revealing that a high abundance of the cellular tRNA^fMet is crucial for the fidelity of initiator tRNA selection on the ribosomal P-site in E. coli. We discuss possible implications of the changes in the cellular tRNA^fMet abundance in proteome remodeling.