Insights into Editing from an Ile-tRNA Synthetase Structure with tRNA Ile and Mupirocin

Abstract

Isoleucyl–transfer RNA (tRNA) synthetase (IleRS) joins Ile to tRNA^Ile at its synthetic active site and hydrolyzes incorrectly acylated amino acids at its editing active site. The 2.2 angstrom resolution crystal structure of Staphylococcus aureus IleRS complexed with tRNA^Ile and Mupirocin shows the acceptor strand of the tRNA^Ile in the continuously stacked, A-form conformation with the 3′ terminal nucleotide in the editing active site. To position the 3′ terminus in the synthetic active site, the acceptor strand must adopt the hairpinned conformation seen in tRNA^Gln complexed with its synthetase. The amino acid editing activity of the IleRS may result from the incorrect products shuttling between the synthetic and editing active sites, which is reminiscent of the editing mechanism of DNA polymerases.