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Sphingolipids with Very Long-chain Fatty Acids Regulate Vacuole Fusion During Tethering and Docking

Logan R. Hurst, Chi Zhang, Thomas D.D. Kazmirchuk, David A. Rivera-Kohr, Christopher L. Brett, View ORCID ProfileRutilio A. Fratti
doi: https://doi.org/10.1101/2020.02.17.953331
Logan R. Hurst
1Department of Biochemistry, and University of Illinois Urbana-Champaign, Urbana, IL, USA
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Chi Zhang
1Department of Biochemistry, and University of Illinois Urbana-Champaign, Urbana, IL, USA
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Thomas D.D. Kazmirchuk
2Department of Biology, Concordia University, Montreal, QC, Canada
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David A. Rivera-Kohr
1Department of Biochemistry, and University of Illinois Urbana-Champaign, Urbana, IL, USA
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Christopher L. Brett
2Department of Biology, Concordia University, Montreal, QC, Canada
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Rutilio A. Fratti
1Department of Biochemistry, and University of Illinois Urbana-Champaign, Urbana, IL, USA
3Center for Biophysics and Quantitative Biology, University of Illinois Urbana-Champaign, Urbana, IL, USA
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  • ORCID record for Rutilio A. Fratti
  • For correspondence: rfratti@illinois.edu
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Abstract

The role of sphingolipids in controlling the endolysosomal membrane trafficking remains unclear. Here, we show that in Saccharomyces cerevisiae sphingolipids containing very long-chain fatty-acids (VLCAs) promote homotypic vacuolar fusion. Yeast that lack the C26 VLCA elongase Elo3p display morphological and vacuolar abnormalities. Vacuoles isolated from these cells displayed reduced levels of in vitro fusion, which we traced to a block in tethering and docking. We found that C26 VLCFA deficient yeast mislocalize fusion markers, and the small GTPases Rho1p and Ypt7p fail to selectively concentrate at the boundary and vertex domains of vacuoles isolated from these yeasts. Surprisingly, we only observed mild changes to the localization of other regulatory lipids, but membrane fluidity and solubility was significantly altered. Taken together, these results suggest that sphingolipids containing C26 VLCFAs act as regulatory lipids in the homotypic vacuolar fusion cascade by assembling membrane microdomains that promote the protein and lipid machinery required for the tethering and docking of vacuoles.

Summary Many sphingolipids contain very-long chain fatty-acids with 26 carbons. The deletion of Elo3, the elongase that adds the final two carbons results in pleiotropic effects that negatively alter membrane fusion at the tethering and docking stages.

Footnotes

  • Abbreviations used: VLCFA, Very long-chain fatty acid; LCB, long-chain base; FAS, fatty acid synthase; PtdInsP, phosphatidylinositol phosphate; IPC, inositolphosphoryl ceramide.

Copyright 
The copyright holder for this preprint is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. All rights reserved. No reuse allowed without permission.
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Posted February 18, 2020.
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Sphingolipids with Very Long-chain Fatty Acids Regulate Vacuole Fusion During Tethering and Docking
Logan R. Hurst, Chi Zhang, Thomas D.D. Kazmirchuk, David A. Rivera-Kohr, Christopher L. Brett, Rutilio A. Fratti
bioRxiv 2020.02.17.953331; doi: https://doi.org/10.1101/2020.02.17.953331
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Sphingolipids with Very Long-chain Fatty Acids Regulate Vacuole Fusion During Tethering and Docking
Logan R. Hurst, Chi Zhang, Thomas D.D. Kazmirchuk, David A. Rivera-Kohr, Christopher L. Brett, Rutilio A. Fratti
bioRxiv 2020.02.17.953331; doi: https://doi.org/10.1101/2020.02.17.953331

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