title: An organized co-assembly of clathrin adaptors is essential for endocytosis.
abstract: Clathrin-mediated endocytosis, the main trafficking route from the plasma membrane to the cytoplasm, is critical to many fundamental cellular processes. Clathrin, coupled to the membrane by adaptor proteins, is thought to play a major structural role in endocytosis by self-assembling into a cage-like lattice around the forming vesicle. Although clathrin adaptors are essential for endocytosis, little is known about their structural role in this process. Here we show that the membrane-binding domains of two conserved clathrin adaptors, Sla2 and Ent1, co-assemble in a PI(4,5)P2-dependent manner to form organized lattices on membranes. We determined the structure of the co-assembled lattice by electron cryo-microscopy and designed mutations that specifically impair the lattice formation in vitro. We show that these mutations block endocytosis in vivo. We suggest that clathrin adaptors not only link the polymerized clathrin to the membrane but also form an oligomeric structure, which is essential for membrane remodeling during endocytosis.
authors.affiliation: Cell Biology and Biophysics Unit, European Molecular Biology Laboratory (EMBL), 69117 Heidelberg, Germany.
authors.forename: Michal
authors.initials: M
authors.lastname: Skruzny
authors.affiliation: Cell Biology and Biophysics Unit, European Molecular Biology Laboratory (EMBL), 69117 Heidelberg, Germany; Structural and Computational Biology Unit, EMBL, 69117 Heidelberg, Germany.
authors.forename: Ambroise
authors.initials: A
authors.lastname: Desfosses
authors.affiliation: Structural and Computational Biology Unit, EMBL, 69117 Heidelberg, Germany.
authors.forename: Simone
authors.initials: S
authors.lastname: Prinz
authors.affiliation: Structural and Computational Biology Unit, EMBL, 69117 Heidelberg, Germany.
authors.forename: Svetlana O
authors.initials: SO
authors.lastname: Dodonova
authors.affiliation: EMBL Hamburg Unit, EMBL, 22607 Hamburg, Germany.
authors.forename: Anna
authors.initials: A
authors.lastname: Gieras
authors.affiliation: WP-79 Sample Environment Group, European XFEL GmbH, 22607 Hamburg, Germany.
authors.forename: Charlotte
authors.initials: C
authors.lastname: Uetrecht
authors.affiliation: Structural and Computational Biology Unit, EMBL, 69117 Heidelberg, Germany; EMBL Hamburg Unit, EMBL, 22607 Hamburg, Germany.
authors.forename: Arjen J
authors.initials: AJ
authors.lastname: Jakobi
authors.affiliation: Cell Biology and Biophysics Unit, European Molecular Biology Laboratory (EMBL), 69117 Heidelberg, Germany.
authors.forename: Marc
authors.initials: M
authors.lastname: Abella
authors.affiliation: Structural and Computational Biology Unit, EMBL, 69117 Heidelberg, Germany.
authors.forename: Wim J H
authors.initials: WJ
authors.lastname: Hagen
authors.affiliation: WP-79 Sample Environment Group, European XFEL GmbH, 22607 Hamburg, Germany.
authors.forename: Joachim
authors.initials: J
authors.lastname: Schulz
authors.affiliation: EMBL Hamburg Unit, EMBL, 22607 Hamburg, Germany.
authors.forename: Rob
authors.initials: R
authors.lastname: Meijers
authors.affiliation: Structural and Computational Biology Unit, EMBL, 69117 Heidelberg, Germany.
authors.forename: Vladimir
authors.initials: V
authors.lastname: Rybin
authors.affiliation: Cell Biology and Biophysics Unit, European Molecular Biology Laboratory (EMBL), 69117 Heidelberg, Germany; Structural and Computational Biology Unit, EMBL, 69117 Heidelberg, Germany.
authors.forename: John A G
authors.initials: JA
authors.lastname: Briggs
authors.affiliation: Structural and Computational Biology Unit, EMBL, 69117 Heidelberg, Germany. Electronic address: carsten.sachse@embl.de.
authors.forename: Carsten
authors.initials: C
authors.lastname: Sachse
authors.affiliation: Cell Biology and Biophysics Unit, European Molecular Biology Laboratory (EMBL), 69117 Heidelberg, Germany; Structural and Computational Biology Unit, EMBL, 69117 Heidelberg, Germany. Electronic address: kaksonen@embl.de.
authors.forename: Marko
authors.initials: M
authors.lastname: Kaksonen
mesh_terms: D033942:Adaptor Proteins, Vesicular Transport / Q000378:metabolism*; D001692:Biological Transport; D002462:Cell Membrane / Q000378:metabolism; D003598:Cytoskeletal Proteins; D004023:Dictyostelium / Q000378:metabolism*; D004705:Endocytosis / Q000502:physiology*; D033721:Equilibrative Nucleoside Transporter 1 / Q000378:metabolism*; D010766:Phosphorylation; D017434:Protein Structure, Tertiary; D022161:Transport Vesicles; D015003:Yeasts / Q000378:metabolism*
chemical_list: D033942:Adaptor Proteins, Vesicular Transport; D003598:Cytoskeletal Proteins; D033721:Equilibrative Nucleoside Transporter 1