Inhibitors of protein synthesis identified by a high throughput multiplexed translation screen
AUTOR(ES)
Novac, Olivia
FONTE
Oxford University Press
RESUMO
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.
ACESSO AO ARTIGO
http://www.pubmedcentral.nih.gov/articlerender.fcgi?artid=373382Documentos Relacionados
- A quantitative, high-throughput screen for protein stability
- Protein expression profiling arrays: tools for the multiplexed high-throughput analysis of proteins
- High-Throughput Screen for Inhibitors of Transglycosylase and/or Transpeptidase Activities of Escherichia coli Penicillin Binding Protein 1b
- Identification of Synthetic Endothelial Cell-Specific Promoters by Use of a High-Throughput Screen
- High-throughput screen for detecting antimycobacterial agents.