Membrane surface charge is critical for the transient, yet specific recruitment of proteins with polybasic regions to certain organelles. In eukaryotes, the plasma membrane (PM) is the most electronegative compartment of the cell, which specifies its identity. As such, membrane electrostatics is a central parameter in signaling, intracellular trafficking, and polarity. Here, we explore which are the lipids that control membrane electrostatics using plants as a model. We show that phosphatidylinositol-4-phosphate (PI4P), phosphatidic acidic (PA), and phosphatidylserine (PS) are separately required to generate the electrostatic signature of the plant PM. In addition, we reveal the existence of an electrostatic territory that is organized as a gradient along the endocytic pathway and is controlled by PS/PI4P combination. Altogether, we propose that combinatorial lipid composition of the cytosolic leaflet of organelles not only defines the electrostatic territory but also distinguishes different functional compartments within this territory by specifying their varying surface charges.

Bordeaux Imaging Center UMS 3420 CNRS US4 INSERM University of Bordeaux Bordeaux 33000 France

Institut Jacques Monod CNRS UMR 7592 Université Paris Diderot Sorbonne Paris Cité Paris 75013 France

Institute of Experimental Botany Czech Academy of Sciences 16502 Prague 6 Lysolaje Czech Republic

Laboratoire Reproduction et Développement des Plantes Université de Lyon ENS de Lyon UCB Lyon 1 CNRS INRA Lyon 69342 France

Metabolome Lipidome Facility of Bordeaux Functional Genomics Center Villenave d'Ornon France

UMR 5200 Membrane Biogenesis Laboratory CNRS University of Bordeaux Bâtiment A3 INRA Bordeaux Aquitaine 71 Avenue Edouard Bourlaux CS 20032 Villenave d'Ornon 33140 France