Published online 9 February 2004
Nucleic Acids Research, 2004, Vol. 32, No. 3 902-915
© 2004 Oxford University Press
Inhibitors of protein synthesis identified by a high throughput multiplexed translation screen
1 Department of Biochemistry, 2 McGill High Throughput Screening Facility and 3 McGill Cancer Center, 3655 Promenade Sir William Osler, McIntyre Medical Sciences Building, McGill University, Montreal, Quebec H3G 1Y6, Canada
*To whom correspondence should be addressed at: McIntyre Medical Sciences Building, Room 810, 3655 Promenade Sir William Osler, McGill University, Montreal, Quebec H3G 1Y6, Canada. Tel: +1 514 398 2323; Fax: +1 514 398 7384; Email: jerry.pelletier{at}mcgill.ca
The use of small molecule inhibitors of cellular processes is a powerful approach to understanding gene function that complements the genetic approach. We have designed a high throughput screen to identify new inhibitors of eukaryotic protein synthesis. We used a bicistronic mRNA reporter to multiplex our assay and simultaneously screen for inhibitors of cap-dependent initiation, internal initiation and translation elongation/termination. Functional screening of >90 000 compounds in an in vitro translation reaction identified 36 inhibitors, 14 of which are known inhibitors of translation and 18 of which are nucleic acid-binding ligands. Our results indicate that intercalators constitute a large class of protein synthesis inhibitors. Four non-intercalating compounds were identified, three of which block elongation and one of which inhibits initiation. The novel inhibitor of initiation affects 5' end-mediated initiation, as well as translation initiated from picornaviral IRESs, but does not significantly affect internal initiation from the hepatitis C virus 5'-untranslated region. This compound should be useful for delineating differences in mechanism of initiation among IRESs.
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