Nucl. Acids Res.-2013-Orelle-e144.pdf (1.85 MB)
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Identifying the targets of aminoacyl-tRNA synthetase inhibitors by primer extension inhibition

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posted on 19.02.2014, 00:00 authored by Cedric Orelle, Teresa Szal, Dorota Klepacki, Karen J. Shaw, Nora Vazquez-Laslop, Alexander S. Mankin
Aminoacyl-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.

Funding

National Institutes of Health [R03 DA035191]; DTRA Chemical/Biological Technologies Directorate contract (in part) [HDTRA1-10-C-004].

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Publisher Statement

© The Author(s) 2013. Published by Oxford University Press. This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/ by-nc/3.0/), which permits non-commercial re-use, distribution, and reproduction in any medium, provided the original work is properly cited. This is an electronic version of an article published in Orelle C, Szal T, Klepacki D, Shaw KJ, Vázquez-Laslop N, Mankin AS. Identifying the targets of aminoacyl-tRNA synthetase inhibitors by primer extension inhibition. Nucleic Acids Research. 2013 Aug;41(14):e144. doi: 10.1093/nar/gkt526.

Publisher

Oxford University Press

Language

en_US

issn

1362-4962

Issue date

01/08/2013

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