Inhibitors of protein synthesis identified by a high throughput multiplexed translation screen

Nucleic Acids Res. 2004 Feb 9;32(3):902-15. doi: 10.1093/nar/gkh235. Print 2004.

Abstract

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.

Publication types

  • Evaluation Study
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Genetic Techniques
  • Mice
  • Peptide Chain Initiation, Translational / drug effects
  • Protein Biosynthesis / drug effects*
  • Protein Synthesis Inhibitors / chemistry
  • Protein Synthesis Inhibitors / pharmacology*
  • Proteins / genetics
  • RNA, Messenger / analysis
  • Rabbits
  • Reproducibility of Results
  • Structure-Activity Relationship

Substances

  • Protein Synthesis Inhibitors
  • Proteins
  • RNA, Messenger