C6 pyridinium ceramide influences alternative pre-mRNA splicing by inhibiting protein phosphatase-1

Chiranthani Sumanasekera, Olga Kelemen, Monique Beullens, Brandon E. Aubol, Joseph A. Adams, Manjula Sunkara, Andrew Morris, Mathieu Bollen, Athena Andreadis, Stefan Stamm

Research output: Contribution to journalArticlepeer-review

20 Scopus citations


Alternative pre-mRNA processing is a central element of eukaryotic gene regulation. The cell frequently alters the use of alternative exons in response to physiological stimuli. Ceramides are lipid-signaling molecules composed of sphingosine and a fatty acid. Previously, water-insoluble ceramides were shown to change alternative splicing and decrease SR-protein phosphorylation by activating protein phosphatase-1 (PP1). To gain further mechanistical insight into ceramide-mediated alternative splicing, we analyzed the effect of C6 pyridinium ceramide (PyrCer) on alternative splice site selection. PyrCer is a water-soluble ceramide analog that is under investigation as a cancer drug. We found that PyrCer binds to the PP1 catalytic subunit and inhibits the dephosphorylation of several splicing regulatory proteins containing the evolutionarily conserved RVxF PP1-binding motif (including PSF/SFPQ, Tra2-beta1 and SF2/ASF). In contrast to natural ceramides, PyrCer promotes phosphorylation of splicing factors. Exons that are regulated by PyrCer have in common suboptimal splice sites, are unusually short and share two 4-nt motifs, GAAR and CAAG. They are dependent on PSF/SFPQ, whose phosphorylation is regulated by PyrCer. Our results indicate that lipids can influence pre-mRNA processing by regulating the phosphorylation status of specific regulatory factors, which is mediated by protein phosphatase activity.

Original languageEnglish
Pages (from-to)4025-4039
Number of pages15
JournalNucleic Acids Research
Issue number9
StatePublished - May 2012

Bibliographical note

Funding Information:
European Alternative Splicing Network of Excellence (EURASNET) (LSHG-CT-2005-518238) and National Institutes of Health (R21HD056195, 2P20 RR020171, P20RR021954 and RO1GM083187, GM50388, GM67969) as well as an ARRA supplement to J.A.A. Funding for open access charge: National Institutes of Health (RO1GM083187).

ASJC Scopus subject areas

  • Genetics


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