Sphingolipids mediate polar sorting of PIN2 through phosphoinositide consumption at the trans-Golgi network

Yoko Ito,Patrick Moreau,Stéphane Claverol,Yohann Boutté,Matthieu Pierre Platre,Yvon Jaillais,Lise C Noack,Laetitia Fouillen,Nicolas Esnay,Louise Fougère,Wilhelm Menzel,Valérie Wattelet-Boyer

doi:10.1038/s41467-021-24548-0

Abstract

The lipid composition of organelles acts as a landmark to define membrane identity and specify subcellular function. Phosphoinositides are anionic lipids acting in protein sorting and trafficking at the trans-Golgi network (TGN). In animal cells, sphingolipids control the turnover of phosphoinositides through lipid exchange mechanisms at endoplasmic reticulum/TGN contact sites. In this study, we discover a mechanism for how sphingolipids mediate phosphoinositide homeostasis at the TGN in plant cells. Using multiple approaches, we show that a reduction of the acyl-chain length of sphingolipids results in an increased level of phosphatidylinositol-4-phosphate (PtdIns(4)P or PI4P) at the TGN but not of other lipids usually coupled to PI4P during exchange mechanisms. We show that sphingolipids mediate Phospholipase C (PLC)-driven consumption of PI4P at the TGN rather than local PI4P synthesis and that this mechanism is involved in the polar sorting of the auxin efflux carrier PIN2 at the TGN. Together, our data identify a mode of action of sphingolipids in lipid interplay at the TGN during protein sorting.

Highlights

The lipid composition of organelles acts as a landmark to define membrane identity and specify subcellular function
The interplay between sphingolipids and phosphoinositides has been evidenced in mammals at ER/trans-Golgi network (TGN) contact sites and involves a complex regulatory homeostatic loop based on OSBPmediated PI4P/sterols or PI4P/PS lipid exchange mechanisms induced by the grafting of a polar head on ceramide at the TGN5,6,10
We cannot completely exclude the possibility that the effect of sphingolipids on PI4P involves lipid exchange mechanisms induced by sphingolipid-metabolic flux, our results strongly suggest that the interplay between sphingolipids and phosphoinositides originates from another mechanism

Summary

Introduction

The lipid composition of organelles acts as a landmark to define membrane identity and specify subcellular function. Sphingolipids control the turnover of phosphoinositides through lipid exchange mechanisms at endoplasmic reticulum/ TGN contact sites. PI4P recruits both CERT, facilitating ceramide transfer, and oxysterol-binding proteins (OSBP), which exchange PI4P for sterols at ER-TGN contact sites[6,18]. Alteration of either the acyl-chain length of sphingolipids or PI4KIIIβ function result in swollen TGN-vesicles being less interconnected with membrane tubules and alteration in TGN-mediated sorting of the auxin efflux carrier PIN2 that localizes in a polar fashion at apical membrane of root epidermal cells, indicating a potential interplay between sphingolipids and phosphoinositides at the TGN in plant cells[22,26]. Our results establish a mode of action of sphingolipids on phosphoinositide homeostasis during protein sorting through another mode of PI4P consumption than the PI4P/ sterols exchange mechanism

Methods

Results

Conclusion