AtEH/Pan1 proteins drive phase separation of the TPLATE complex and clathrin polymerisation during plant endocytosis

Dragwidge, J. M.; Wang, Y.; Brocard, L.; De Meyer, A.; Hudecek, R.; Eeckhout, D.; Chambaud, C.; Pejchar, P.; Potocky, M.; Vandorpe, M.; Bernard, A.; De Jaeger, G.; Pleskot, R.; Fang, X.; Van Damme, D.

doi:10.1101/2022.03.17.484738

bioRxiv · 10.1101/2022.03.17.484738

AtEH/Pan1 proteins drive phase separation of the TPLATE complex and clathrin polymerisation during plant endocytosis

Dragwidge, J. M.·Wang, Y.·Brocard, L.·De Meyer, A.·Hudecek, R.·Eeckhout, D.·Chambaud, C.·Pejchar, P.·Potocky, M.·Vandorpe, M.·Bernard, A.·De Jaeger, G.·Pleskot, R.·Fang, X.·Van Damme, D.

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Abstract

Clathrin-mediated endocytosis (CME) is an essential cellular internalisation pathway involving the dynamic assembly of clathrin and accessory proteins to form membrane-bound vesicles. In plants, the evolutionarily ancient TSET/TPLATE complex (TPC) plays an essential, but not well-defined role in CME. 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, early-, and late-stage endocytic accessory proteins. Importantly, clathrin forms ordered assemblies within the condensate environment. Biomolecular condensation therefore acts to promote dynamic protein assemblies throughout clathrin-mediated endocytosis. Furthermore, the disordered region sequence properties of AtEH1 regulate the material properties of the endocytic condensates in vivo and alteration of these material properties influences endocytosis dynamics, and consequently plant adaptive growth. HighlightsO_LIAtEH subunits are endocytic scaffolds which drive condensation of the TPC C_LIO_LIAtEH1 condensates nucleate on the plasma membrane via lipid interactions C_LIO_LICondensation of AtEH1/TPC facilitates clathrin re-arrangement and assembly C_LIO_LIAtEH1 IDR1 composition controls condensate properties to regulate endocytosis C_LI

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Dragwidge, J. M., Wang, Y., Brocard, L., De Meyer, A., Hudecek, R., Eeckhout, D., Chambaud, C., Pejchar, P., Potocky, M., Vandorpe, M., Bernard, A., De Jaeger, G., Pleskot, R., Fang, X., Van Damme, D.. 2022-03-17. AtEH/Pan1 proteins drive phase separation of the TPLATE complex and clathrin polymerisation during plant endocytosis. https://doi.org/10.1101/2022.03.17.484738

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