Nucl. Acids Res.-2013-Orelle-e144.pdf (1.85 MB)
Download fileIdentifying the targets of aminoacyl-tRNA synthetase inhibitors by primer extension inhibition
journal contribution
posted on 2014-02-19, 00:00 authored by Cedric Orelle, Teresa Szal, Dorota Klepacki, Karen J. Shaw, Nora Vazquez-Laslop, Alexander S. MankinAminoacyl-transfer RNA (tRNA) synthetases (RS) are
essential components of the cellular translation machinery
and can be exploited for antibiotic discovery.
Because cells have many different RS, usually
one for each amino acid, identification of the
specific enzyme targeted by a new natural or synthetic
inhibitor can be cumbersome. We describe
the use of the primer extension technique in conjunction
with specifically designed synthetic genes
to identify the RS targeted by an inhibitor.
Suppression of a synthetase activity reduces the
amount of the cognate aminoacyl-tRNA in a cellfree
translation system resulting in arrest of translation
when the corresponding codon enters the
decoding center of the ribosome. The utility of the
technique is demonstrated by identifying a switch in
target specificity of some synthetic inhibitors of
threonyl-tRNA synthetase.