Loss of hydroxyl groups from the ceramide moiety can modify the lateral diffusion of membrane proteins in S. cerevisiae

Satoshi Uemura, Fumi Shishido, Motohiro Tani, Takahiro Mochizuki, Fumiyoshi Abe, Jin Ichi Inokuchi

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15 Citations (Scopus)

Abstract

In the yeast Saccharomyces cerevisiae , structural diversities of complex sphingolipids [inositol phosphorylceramide (IPC), mannosylinositol phosphorylceramide, and mannosyldiinositol phosphorylceramide] are often observed in the presence or absence of hydroxyl groups on the C-4 position of long-chain base (C4-OH) and the C-2 position of very long-chain fatty acids (C2-OH), but the biological signifi - cance of these groups remains unclear. Here, we evaluated cellular membrane fl uidity in hydroxyl group-defective yeast mutants by fl uorescence recovery after photobleaching. The lateral diffusion of enhanced green fl uorescent proteintagged hexose transporter 1 (Hxt1-EGFP) was infl uenced by the absence of C4-OH and/or C2-OH. Notably, the fl uorescence recovery of Hxt1-EGFP was dramatically decreased in the sur2 - mutant (absence of C4-OH) under the csg1 - csh1 - background, in which mannosylation of IPC is blocked leading to IPC accumulation, while the recovery in the scs7 - mutant (absence of C2-OH) under the same background was modestly decreased. In addition, the amount of low affi nity tryptophan transporter 1 (Tat1)-EGFP was markedly decreased in the sur2 - csg1 - csh1 - mutant and accumulated in intracellular membranes in the scs7 - csg1 - csh1 - mutant without altering its protein expression. These results suggest that C4-OH and C2-OH are most probably critical factors for maintaining membrane fl uidity and proper turnover of membrane molecules in yeast containing complex sphingolipids with only one hydrophilic head group. -Uemura, S., F. Shishido, M. Tani, T. Mochizuki, F. Abe, and J-i. Inokuchi. Loss of hydroxyl groups from the ceramide moiety can modify the lateral diffusion of membrane proteins in S. cerevisiae . J. Lipid Res. 2014. 55: 1343 - 1356 .

Original languageEnglish
Pages (from-to)1343-1356
Number of pages14
JournalJournal of Lipid Research
Volume55
Issue number7
DOIs
Publication statusPublished - Jul 2014

All Science Journal Classification (ASJC) codes

  • Biochemistry
  • Endocrinology
  • Cell Biology

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