Inhibitors of protein synthesis identified by a high throughput multiplexed translation screen.
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.
Novac
Olivia
O
Department of Biochemistry, McGill Cancer Center, 3655 Promenade Sir William Osler, McIntyre Medical Sciences Building, McGill University, Montreal, Quebec H3G 1Y6, Canada.
Guenier
Anne-Sophie
AS
Pelletier
Jerry
J
D000818:Animals; D005821:Genetic Techniques; D051379:Mice; D010442:Peptide Chain Initiation, Translational / Q000187:drug effects; D014176:Protein Biosynthesis / Q000187:drug effects*; D011500:Protein Synthesis Inhibitors / Q000737:chemistry / Q000494:pharmacology*; D011506:Proteins / Q000235:genetics; D012333:RNA, Messenger / Q000032:analysis; D011817:Rabbits; D015203:Reproducibility of Results; D013329:Structure-Activity Relationship
D011500:Protein Synthesis Inhibitors; D011506:Proteins; D012333:RNA, Messenger