Identification of small molecule inhibitors of pre-mRNA splicing

Andrea Pawellek, Stuart McElroy, Timur Samatov, Lee Mitchell, Andrew Woodland, Ursula Ryder, David Gray, Reinhard Lührmann, Angus I. Lamond (Lead / Corresponding author)

Research output: Contribution to journalArticle

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Abstract

Eukaryotic pre-mRNA splicing is an essential step in gene expression for all genes that contain introns. In contrast with transcription and translation, few well-characterized chemical inhibitors are available with which to dissect the splicing process, particularly in cells. Therefore, the identification of specific, small molecules that either inhibit, or modify, pre-mRNA splicing would be valuable for research and potentially also for therapeutic applications. We have screened a highly curated library of 71,504 drug-like small molecules using a high throughput in vitro splicing assay. This identified 10 new compounds that both inhibit pre-mRNA splicing in vitro and modify splicing of endogenous pre-mRNA in cells. One of these splicing modulators, DDD00107587 (termed "madrasin" i.e., (2-((7methoxy-4-methylquinazolin-2-yl)amino)-5,6-dimethylpyrimidin-4(3H)-one RNA splicing inhibitor), was studied in more detail. Madrasin interferes with the early stages of spliceosome assembly and stalls spliceosome assembly at the A complex. Madrasin is cytotoxic at higher concentrations, while at lower concentrations it induces cell cycle arrest, promotes a specific reorganization of subnuclear protein localization and modulates splicing of multiple pre-mRNAs in both HeLa and HEK293 cells.
Original languageEnglish
Pages (from-to)34683-34698
Number of pages16
JournalJournal of Biological Chemistry
Volume289
Issue number50
Early online date3 Oct 2014
DOIs
Publication statusPublished - 12 Dec 2014

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