title: Charged multivesicular body protein 2B (CHMP2B) of the endosomal sorting complex required for transport-III (ESCRT-III) polymerizes into helical structures deforming the plasma membrane.
abstract: The endosomal sorting complexes required for transport (ESCRT-0-III) allow membrane budding and fission away from the cytosol. This machinery is used during multivesicular endosome biogenesis, cytokinesis, and budding of some enveloped viruses. Membrane fission is catalyzed by ESCRT-III complexes made of polymers of charged multivesicular body proteins (CHMPs) and by the AAA-type ATPase VPS4. How and which of the ESCRT-III subunits sustain membrane fission from the cytoplasmic surface remain uncertain. In vitro, CHMP2 and CHMP3 recombinant proteins polymerize into tubular helical structures, which were hypothesized to drive vesicle fission. However, this model awaits the demonstration that such structures exist and can deform membranes in cellulo. Here, we show that depletion of VPS4 induces specific accumulation of endogenous CHMP2B at the plasma membrane. Unlike other CHMPs, overexpressed full-length CHMP2B polymerizes into long, rigid tubes that protrude out of the cell. CHMP4s relocalize at the base of the tubes, the formation of which depends on VPS4. Cryo-EM of the CHMP2B membrane tubes demonstrates that CHMP2B polymerizes into a tightly packed helical lattice, in close association with the inner leaflet of the membrane tube. This association is tight enough to deform the lipid bilayer in cases where the tubular CHMP2B helix varies in diameter or is closed by domes. Thus, our observation that CHMP2B polymerization scaffolds membranes in vivo represents a first step toward demonstrating its structural role during outward membrane deformation.
authors.affiliation: INSERM, U836, Equipe 2, Neurodégénérescence et Plasticité, Grenoble F-38042, France.
authors.forename: Gilles
authors.initials: G
authors.lastname: Bodon
authors.forename: Romain
authors.initials: R
authors.lastname: Chassefeyre
authors.forename: Karin
authors.initials: K
authors.lastname: Pernet-Gallay
authors.forename: Nicolas
authors.initials: N
authors.lastname: Martinelli
authors.forename: Grégory
authors.initials: G
authors.lastname: Effantin
authors.forename: David Lutje
authors.initials: DL
authors.lastname: Hulsik
authors.forename: Agnès
authors.initials: A
authors.lastname: Belly
authors.forename: Yves
authors.initials: Y
authors.lastname: Goldberg
authors.forename: Christine
authors.initials: C
authors.lastname: Chatellard-Causse
authors.forename: Béatrice
authors.initials: B
authors.lastname: Blot
authors.forename: Guy
authors.initials: G
authors.lastname: Schoehn
authors.forename: Winfried
authors.initials: W
authors.lastname: Weissenhorn
authors.forename: Rémy
authors.initials: R
authors.lastname: Sadoul
mesh_terms: D002462:Cell Membrane / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D056827:Endosomal Sorting Complexes Required for Transport / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D011992:Endosomes / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D006367:HeLa Cells; D006801:Humans; D055503:Protein Multimerization / Q000502:physiology*; D020836:Protein Structure, Quaternary
chemical_list: C503812:CHMP2B protein, human; D056827:Endosomal Sorting Complexes Required for Transport