@article{66375ca429dc40a792bccd48f508f20b,
title = "Coupling between endocytosis and sphingosine kinase 1 recruitment",
abstract = "Genetic studies have suggested a functional link between cholesterol/sphingolipid metabolism and endocytic membrane traffic. Here we show that perturbing the cholesterol/sphingomyelin balance in the plasma membrane results in the massive formation of clusters of narrow endocytic tubular invaginations positive for N-BAR proteins. These tubules are intensely positive for sphingosine kinase 1 (SPHK1). SPHK1 is also targeted to physiologically occurring early endocytic intermediates, and is highly enriched in nerve terminals, which are cellular compartments specialized for exo/endocytosis. Membrane recruitment of SPHK1 involves a direct, curvature-sensitive interaction with the lipid bilayer mediated by a hydrophobic patch on the enzymes surface. The knockdown of SPHKs results in endocytic recycling defects, and a mutation that disrupts the hydrophobic patch of Caenorhabditis elegans SPHK fails to rescue the neurotransmission defects in loss-of-function mutants of this enzyme. Our studies support a role for sphingosine phosphorylation in endocytic membrane trafficking beyond the established function of sphingosine-1-phosphate in intercellular signalling.",
author = "Hongying Shen and Francesca Giordano and Yumei Wu and Jason Chan and Chen Zhu and Ira Milosevic and Xudong Wu and Kai Yao and Bo Chen and Tobias Baumgart and Derek Sieburth and \{De Camilli\}, Pietro",
note = "Funding Information: Experiments in C. elegans. All behavioural and microscopy experiments were performed on young adult hermaphrodites. sphk-1(ok1097) was provided by the Caenorhabditis Genetics Center, which is funded by NIH Office of Research Infrastructure Programs (P40 OD010440). The wild-type reference strain was N2 Bristol. sphk-1(ok1097) was outcrossed 10 times before use in experiments. The following transgenic strains were also used (strain number (extrachromosomal array; plasmid)): OJ889 (vjEx338; pJC35), JPC01 (jcaEx01; pJC152) and OJ2111 (vjEx804; pDS449). Funding Information: We thank L. Liang and J. Duncan (Yale University) for help with the automatic tracking of clathrin-coated pits dynamics, S. Ferguson, A. Frost and T. Walther for discussion and advice, J. Baskin for thorough reading of the manuscript, F. Wilson, L. Lucast, L. Liu and H. Czapla for outstanding technical support, M. Graham, X. Liu and S. Wilson for help with microscopy experiments, and members of T. Walther, T. Melia and C. Burd laboratories (Yale University) for help with lipid experiments. We also acknowledge the help of the Yale Center for Cellular and Molecular Imaging and Yale Center for Genomics and Proteomics. This work was supported in part by grants from the NIH (NS36251, DK45735 and DA018343 to P.D.C., GM097552 to T.B., and NS071085 to D.S.) and from the Ellison Medical Foundation to P.D.C.",
year = "2014",
month = jul,
doi = "10.1038/ncb2987",
language = "English",
volume = "16",
pages = "652--662",
journal = "Nature Cell Biology",
issn = "1465-7392",
publisher = "Nature Research",
number = "7",
}