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Phosphorylated glycosphingolipids essential for cholesterol mobilization in Caenorhabditis elegans

Nature Chemical Biology volume 13, pages 647–654 (2017)Cite this article

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Abstract

The nematode Caenorhabditis elegans requires exogenous cholesterol to survive and its depletion leads to early developmental arrest. Thus, tight regulation of cholesterol storage and distribution within the organism is indispensable. Here, we present a novel class of C. elegans phosphorylated glycosphingolipids, phosphoethanolamine glucosylceramides (PEGCs), capable of rescuing larval arrest induced by sterol starvation. We describe the total synthesis of a major PEGC species and demonstrate that the PEGC synthetic counterpart suppresses the dauer-constitutive phenotype of Niemann–Pick C1 (NPC1) and DAF-7/TGF-β mutant worms caused by impaired intracellular sterol trafficking. PEGC biosynthesis depends on functional NPC1 and TGF-β, indicating that these proteins control larval development at least partly through PEGC. Furthermore, glucosylceramide deficiency dramatically reduced PEGC amounts. However, the resulting developmental arrest could be rescued by oversaturation of food with cholesterol. Taken together, these data show that PEGC is essential for C. elegans development through its regulation of sterol mobilization.

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Figure 1: L2* arrest caused by sterol depletion can be reversed by the addition of a hydrophilic fraction derived from a total lipid extract of worms.
Figure 2: Assigned structure of the novel phosphoglycolipid class PEGC and total synthesis of d17iso-GlcCer (11) and mmPEGC-C22 (1).
Figure 3: PEGC promotes the development of worms by mobilizing internal sterol pools.
Figure 4: Feeding of NPC-1 mutant worms with 1 suppresses Daf-c phenotype.
Figure 5: Mobilization of the internal pool of sterols by PEGC is under the control of TGF-β/DAF-7.
Figure 6: Larval arrest of PEGC-deficient worms (elo-5 and cgt-3;cgt-1) can be rescued by a high-cholesterol diet.

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Acknowledgements

We thank all members of the Kurzchalia group and the Knölker group for helpful discussions and E. Entchev and Ü. Coskun for critical reading of the manuscript. We are grateful to K. Ploß from the Max-Planck Institute for Chemical Ecology for her support through the HPLC–MS experiments. We are grateful to the Caenorhabditis Genetics Center (CGC), A. Antebi, G. Lesa and to H. Riezman for providing worm strains. We are indebted to R. Saini (Department of Chemistry, Technische Universität Dresden) for the synthesis of Δ1,7-DA and lophanol. We thank A. Shevchenko for giving us access to the high-resolution mass spectrometer. We thank A.W. Schmidt and M. Gruner for support. Work in the Knölker laboratory was supported by the European Science Foundation EuroMembrane Network (Deutsche Forschungsgemeinschaft (DFG) grant KN 240/13-1). U.S. is grateful to the TUD Graduate Academy for a fellowship.

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  1. Sebastian Boland and Ulrike Schmidt: These authors contributed equally to this work.

Authors and Affiliations

  1. MPI of Molecular Cell Biology and Genetics, Dresden, Germany

    Sebastian Boland, Vyacheslav Zagoriy, Julio L Sampaio, Jakob Reimann, Sider Penkov & Teymuras V Kurzchalia

  2. Department of Chemistry, Technische Universität Dresden, Dresden, Germany

    Ulrike Schmidt, Raphael F Fritsche, Regina Czerwonka, Tilo Lübken, Jakob Reimann & Hans-Joachim Knölker

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Contributions

S.B., V.Z., S.P., J.R. and T.V.K. designed and performed biological research; S.B., U.S., V.Z., J.L.S. and T.L. performed analytical research; U.S., R.F.F., R.C. and H.-J.K. developed and performed the total synthesis of mmPEGC-C22 and d17iso-GlcCer; S.B., J.L.S., U.S., T.L., H.-J.K. and T.V.K. analyzed data; all authors discussed the data; and S.B., U.S., S.P., H.-J.K. and T.V.K. wrote the paper.

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Correspondence to Hans-Joachim Knölker or Teymuras V Kurzchalia.

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Supplementary Results and Supplementary Figures 1–11 (PDF 2742 kb)

Supplementary Note

Experimental procedures for the syntheses of mmPEGC-C22 and d17iso-GlcCer and corresponding NMR spectra (PDF 5096 kb)

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Boland, S., Schmidt, U., Zagoriy, V. et al. Phosphorylated glycosphingolipids essential for cholesterol mobilization in Caenorhabditis elegans. Nat Chem Biol 13, 647–654 (2017). https://doi.org/10.1038/nchembio.2347

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