TY - JOUR
T1 - Pyrvinium pamoate changes alternative splicing of the serotonin receptor 2C by influencing its RNA structure
AU - Shen, Manli
AU - Bellaousov, Stanislav
AU - Hiller, Michael
AU - De La Grange, Pierre
AU - Creamer, Trevor P.
AU - Malina, Orit
AU - Sperling, Ruth
AU - Mathews, David H.
AU - Stoilov, Peter
AU - Stamm, Stefan
PY - 2013/4
Y1 - 2013/4
N2 - 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.
AB - 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.
UR - http://www.scopus.com/inward/record.url?scp=84876048680&partnerID=8YFLogxK
U2 - 10.1093/nar/gkt063
DO - 10.1093/nar/gkt063
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C2 - 23393189
AN - SCOPUS:84876048680
SN - 0305-1048
VL - 41
SP - 3819
EP - 3832
JO - Nucleic Acids Research
JF - Nucleic Acids Research
IS - 6
ER -