1. Regulation ofChlamydomonasflagella and ependymal cell motile cilia by ceramide-mediated translocation of GSK3
- Author
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Gu Zhu, Guanghu Wang, Jacek Bielawski, Michael B. Dinkins, Qian He, Stefka D. Spassieva, Kara Hardin, Ji Na Kong, Erhard Bieberich, and Tarik Mujadzic
- Subjects
Ceramide ,Serine C-Palmitoyltransferase ,Flagellum ,Ceramides ,Glycogen Synthase Kinase 3 ,Mice ,chemistry.chemical_compound ,Tubulin ,Ependyma ,Ciliogenesis ,Myriocin ,Animals ,Humans ,Cilia ,Molecular Biology ,Cells, Cultured ,biology ,Cilium ,Chlamydomonas ,Acetylation ,Articles ,Cell Biology ,biology.organism_classification ,Sphingolipid ,Cell biology ,Mice, Inbred C57BL ,Biochemistry ,chemistry ,Flagella ,Membrane Trafficking ,Motile cilium ,lipids (amino acids, peptides, and proteins) - Abstract
Cilia are important organelles formed by cell membrane protrusions; however, little is known about their regulation by membrane lipids. A novel, evolutionarily conserved activation mechanism for GSK3 by the sphingolipid (phyto)ceramide is characterized that is critical for ciliogenesis in Chlamydomonas and murine ependymal cells., Cilia are important organelles formed by cell membrane protrusions; however, little is known about their regulation by membrane lipids. We characterize a novel activation mechanism for glycogen synthase kinase-3 (GSK3) by the sphingolipids phytoceramide and ceramide that is critical for ciliogenesis in Chlamydomonas and murine ependymal cells, respectively. We show for the first time that Chlamydomonas expresses serine palmitoyl transferase (SPT), the first enzyme in (phyto)ceramide biosynthesis. Inhibition of SPT in Chlamydomonas by myriocin led to loss of flagella and reduced tubulin acetylation, which was prevented by supplementation with the precursor dihydrosphingosine. Immunocytochemistry showed that (phyto)ceramide was colocalized with phospho–Tyr-216-GSK3 (pYGSK3) at the base and tip of Chlamydomonas flagella and motile cilia in ependymal cells. The (phyto)ceramide distribution was consistent with that of a bifunctional ceramide analogue UV cross-linked and visualized by click-chemistry–mediated fluorescent labeling. Ceramide depletion, by myriocin or neutral sphingomyelinase deficiency (fro/fro mouse), led to GSK3 dephosphorylation and defective flagella and cilia. Motile cilia were rescued and pYGSK3 localization restored by incubation of fro/fro ependymal cells with exogenous C24:1 ceramide, which directly bound to pYGSK3. Our findings suggest that (phyto)ceramide-mediated translocation of pYGSK into flagella and cilia is an evolutionarily conserved mechanism fundamental to the regulation of ciliogenesis.
- Published
- 2015