Pyrvinium pamoate changes alternative splicing of the serotonin receptor 2C by influencing its RNA structure

Authors

Manli Shen, University of KentuckyFollow
Stanislav Bellaousov, University of Rochester
Michael Hiller, Max Planck Institute of Molecular Cell Biology and Genetics, Germany
Pierre de La Grange, Hôpital Saint-Louis, France
Trevor O. Creamer, University of KentuckyFollow
Orit Malina, The Hebrew University of Jerusalem, Israel
Ruth Sperling, The Hebrew University of Jerusalem, Israel
David H. Mathews, University of Rochester
Peter Stoilov, West Virginia University
Stefan Stamm, University of KentuckyFollow

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.

Document Type

Article

Publication Date

2-7-2013

Notes/Citation Information

Published in Nucleic Acids Research, v. 41, no. 6, p. 3819-3832.

© The Author(s) 2013. Published by Oxford University Press.

This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/3.0/), which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.

Repository Citation

Shen, Manli; Bellaousov, Stanislav; Hiller, Michael; de La Grange, Pierre; Creamer, Trevor O.; Malina, Orit; Sperling, Ruth; Mathews, David H.; Stoilov, Peter; and Stamm, Stefan, "Pyrvinium pamoate changes alternative splicing of the serotonin receptor 2C by influencing its RNA structure" (2013). Molecular and Cellular Biochemistry Faculty Publications. 34.
https://uknowledge.uky.edu/biochem_facpub/34