Article Dans Une Revue BioRxiv Année : 2022

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

Jonathan Michael Dragwidge
  • Fonction : Auteur
Yanning Wang
  • Fonction : Auteur
Lysiane Brocard
Andreas de Meyer
  • Fonction : Auteur
Roman Hudeček
  • Fonction : Auteur
Dominique Eeckhout
  • Fonction : Auteur
Clément Chambaud
Přemysl Pejchar
Martin Potocký
  • Fonction : Auteur
Michael Vandorpe
  • Fonction : Auteur
Amelie Bernard
Geert de Jaeger
Roman Pleskot
  • Fonction : Auteur
Xiaofeng Fang
  • Fonction : Auteur
Daniël van Damme

Résumé

Endocytosis is the process by which cells internalise molecules from their cell surface via plasma membrane-derived vesicles. In plants, clathrin-mediated endocytosis requires the evolutionarily ancient TSET/TPLATE complex (TPC), which was lost in metazoan and fungal lineages. TPC is required for membrane bending, but how TPC functions in the initiation of endocytosis and clathrin assembly is unclear. Here we used live-cell imaging and biochemical approaches to investigate the function of the Arabidopsis thaliana TPC subunit AtEH1/Pan1. Using in vitro and in vivo experiments we found that AtEH/Pan1 proteins can self-assemble into condensates through phase separation, which is influenced by both structured and intrinsically disordered regions. The proteome composition of these condensates revealed many key endocytic components which are selectively recruited via prion-likeand IDR-based interactions, including the ESCRT-0 TOM-Like proteins. Furthermore, AtEH/Pan1 condensates selectively nucleate on the plasma membrane by binding specific phospholipid species that are recognised by their EH domains. Visualization of the ultrastructure of the endocytic condensates via CLEM-ET revealed that the coat protein clathrin can assemble into lattices within condensates. Our results reveal that AtEH/Pan1 proteins act as scaffolds to direct endocytic machinery to specific plasma membrane regions to initiate internalisation. These findings provide new insight into the interplay between membranes and protein condensates.
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Dates et versions

hal-03847415 , version 1 (10-11-2022)

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Jonathan Michael Dragwidge, Yanning Wang, Lysiane Brocard, Andreas de Meyer, Roman Hudeček, et al.. AtEH/Pan1 proteins drive phase separation of the TPLATE complex and clathrin polymerisation during plant endocytosis. BioRxiv, 2022, ⟨10.1101/2022.03.17.484738⟩. ⟨hal-03847415⟩

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