title: Pyrvinium pamoate changes alternative splicing of the serotonin receptor 2C by influencing its RNA structure.
abstract: The serotonin receptor 2C plays a central role in mood and appetite control. It undergoes pre-mRNA editing as well as alternative splicing. The RNA editing suggests that the pre-mRNA forms a stable secondary structure in vivo. To identify substances that promote alternative exons inclusion, we set up a high-throughput screen and identified pyrvinium pamoate as a drug-promoting exon inclusion without editing. Circular dichroism spectroscopy indicates that pyrvinium pamoate binds directly to the pre-mRNA and changes its structure. SHAPE (selective 2'-hydroxyl acylation analysed by primer extension) assays show that part of the regulated 5'-splice site forms intramolecular base pairs that are removed by this structural change, which likely allows splice site recognition and exon inclusion. Genome-wide analyses show that pyrvinium pamoate regulates >300 alternative exons that form secondary structures enriched in A-U base pairs. Our data demonstrate that alternative splicing of structured pre-mRNAs can be regulated by small molecules that directly bind to the RNA, which is reminiscent to an RNA riboswitch.
authors: Department of Molecular and Cellular Biochemistry, University of Kentucky, 741 South Limestone, Lexington, KY 40536, USA.
authors: Manli
authors: M
authors: Shen
authors: Stanislav
authors: S
authors: Bellaousov
authors: Michael
authors: M
authors: Hiller
authors: Pierre
authors: P
authors: de La Grange
authors: Trevor P
authors: TP
authors: Creamer
authors: Orit
authors: O
authors: Malina
authors: Ruth
authors: R
authors: Sperling
authors: David H
authors: DH
authors: Mathews
authors: Peter
authors: P
authors: Stoilov
authors: Stefan
authors: S
authors: Stamm
mesh_terms: D017398:Alternative Splicing / Q000187:drug effects*; D001483:Base Sequence; D005091:Exons; D057809:HEK293 Cells; D057166:High-Throughput Screening Assays; D006801:Humans; D008969:Molecular Sequence Data; D009690:Nucleic Acid Conformation; D010802:Phylogeny; D011774:Pyrvinium Compounds / Q000378:metabolism / Q000494:pharmacology*; D017393:RNA Editing; D012322:RNA Precursors / Q000378:metabolism; D012330:RNA, Double-Stranded / Q000737:chemistry / Q000187:drug effects; D012333:RNA, Messenger / Q000737:chemistry / Q000187:drug effects* / Q000378:metabolism; D044405:Receptor, Serotonin, 5-HT2C / Q000235:genetics* / Q000378:metabolism; D017412:Ribonucleoprotein, U1 Small Nuclear / Q000378:metabolism; D017381:Spliceosomes / Q000378:metabolism
chemical_list: D011774:Pyrvinium Compounds; D012322:RNA Precursors; D012330:RNA, Double-Stranded; D012333:RNA, Messenger; D044405:Receptor, Serotonin, 5-HT2C; D017412:Ribonucleoprotein, U1 Small Nuclear; C024631:pyrvinium
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