title: Identification of small molecule inhibitors of pre-mRNA splicing.
abstract: Eukaryotic pre-mRNA splicing is an essential step in gene expression for all genes that contain introns. In contrast to 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 RNAsplicing 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, although 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.
authors.affiliation: From the Centre of Gene Regulation and Expression and.
authors.forename: Andrea
authors.initials: A
authors.lastname: Pawellek
authors.affiliation: the Drug Discovery Unit, University of Dundee, Dundee DD1 5EH, Scotland, United Kingdom and.
authors.forename: Stuart
authors.initials: S
authors.lastname: McElroy
authors.affiliation: the Max Planck Institute for Biophysical Chemistry, 37077 Göttingen, Germany.
authors.forename: Timur
authors.initials: T
authors.lastname: Samatov
authors.affiliation: the Drug Discovery Unit, University of Dundee, Dundee DD1 5EH, Scotland, United Kingdom and.
authors.forename: Lee
authors.initials: L
authors.lastname: Mitchell
authors.affiliation: the Drug Discovery Unit, University of Dundee, Dundee DD1 5EH, Scotland, United Kingdom and.
authors.forename: Andrew
authors.initials: A
authors.lastname: Woodland
authors.affiliation: From the Centre of Gene Regulation and Expression and.
authors.forename: Ursula
authors.initials: U
authors.lastname: Ryder
authors.affiliation: the Drug Discovery Unit, University of Dundee, Dundee DD1 5EH, Scotland, United Kingdom and.
authors.forename: David
authors.initials: D
authors.lastname: Gray
authors.affiliation: the Max Planck Institute for Biophysical Chemistry, 37077 Göttingen, Germany.
authors.forename: Reinhard
authors.initials: R
authors.lastname: Lührmann
authors.affiliation: From the Centre of Gene Regulation and Expression and a.i.lamond@dundee.ac.uk.
authors.forename: Angus I
authors.initials: AI
authors.lastname: Lamond
mesh_terms: D002467:Cell Nucleus / Q000187:drug effects / Q000235:genetics; D004353:Drug Evaluation, Preclinical; D057809:HEK293 Cells; D006367:HeLa Cells; D057166:High-Throughput Screening Assays; D006801:Humans; D011799:Quinazolines / Q000737:chemistry / Q000494:pharmacology; D012322:RNA Precursors / Q000235:genetics*; D012326:RNA Splicing / Q000187:drug effects*; D012333:RNA, Messenger / Q000235:genetics; D054852:Small Molecule Libraries / Q000737:chemistry / Q000494:pharmacology*
chemical_list: D011799:Quinazolines; D012322:RNA Precursors; D012333:RNA, Messenger; D054852:Small Molecule Libraries
keywords: DDD00107587; High Throughput Screening; Madrasin; RNA; RNA Splicing; Small Molecule; Spliceosome