Clathrin-mediated endocytosis is an essential cellular internalization pathway involving the dynamic assembly of clathrin and accessory proteins to form membrane-bound vesicles. The evolutionarily ancient TSET-TPLATE complex (TPC) plays an essential, but ill-defined role in endocytosis in plants. Here we show that two highly disordered TPC subunits, AtEH1 and AtEH2, function as scaffolds to drive biomolecular condensation of the complex. These condensates specifically nucleate on the plasma membrane through interactions with anionic phospholipids, and facilitate the dynamic recruitment and assembly of clathrin, as well as early- and late-stage endocytic accessory proteins. Importantly, condensation promotes ordered clathrin assemblies. TPC-driven biomolecular condensation thereby facilitates dynamic protein assemblies throughout clathrin-mediated endocytosis. Furthermore, we show that a disordered region of AtEH1 controls the material properties of endocytic condensates in vivo. Alteration of these material properties disturbs the recruitment of accessory proteins, influences endocytosis dynamics and impairs plant responsiveness. Our findings reveal how collective interactions shape endocytosis.

Bordeaux Imaging Center INSERM CNRS Université de Bordeaux Bordeaux France

Center for Plant Biology School of Life Sciences Tsinghua University Beijing China

Department of Experimental Plant Biology Faculty of Sciences Charles University Prague Czech Republic

Department of Plant Biotechnology and Bioinformatics Ghent University Ghent Belgium

Institute of Experimental Botany of the Czech Academy of Sciences Prague Czech Republic

Laboratoire de Biogenèse Membranaire CNRS Université de Bordeaux Bordeaux France

VIB Center for Plant Systems Biology Ghent Belgium

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Guidelines for naming and studying plasma membrane domains in plants