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.
|Number of pages||14|
|Journal||Nucleic Acids Research|
|State||Published - Apr 2013|
Bibliographical noteFunding Information:
NIH [RO1 GM083187, P20RR020171; R01 GM076485 to D.H.M.]; Foundation for Prader–Willi Research (to S.S.); Binational Science Foundation (BSF), USA-Israel, Transformative Grant [2010508 to S.S. and R.S.]. Funding for open access charge: NIH [RO1 GM083187].
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