title: Inhibition of ESCRT-II-CHMP6 interactions impedes cytokinetic abscission and leads to cell death.
abstract: Recently the ESCRT-III filamentous complex was designated as the driving force for mammalian cell abscission, that is, fission of the intercellular membrane bridge connecting daughter cells at the end of cytokinesis. However, how ESCRT-III is activated to set on abscission has not been resolved. Here we revisit the role of the upstream canonical ESCRT players ESCRT-II and CHMP6 in abscission. Using high-resolution imaging, we show that these proteins form highly ordered structures at the intercellular bridge during abscission progression. Furthermore, we demonstrate that a truncated version of CHMP6, composed of its first 52 amino acids (CHMP6-N), arrives at the intercellular bridge, blocks abscission, and subsequently leads to cell death. This phenotype is abolished in a mutated version of CHMP6-N designed to prevent CHMP6-N binding to its ESCRT-II partner. Of interest, deleting the first 10 amino acids from CHMP6-N does not interfere with its arrival at the intercellular bridge but almost completely abolishes the abscission failure phenotype. Taken together, these data suggest an active role for ESCRT-II and CHMP6 in ESCRT-mediated abscission. Our work advances the mechanistic understanding of ESCRT-mediated membrane fission in cells and introduces an easily applicable tool for upstream inhibition of the ESCRT pathway in live mammalian cells.
authors.affiliation: Department of Life Sciences and the National Institute for Biotechnology in the Negev (NIBN), Ben-Gurion University of the Negev, Beer-Sheva 84105, Israel.
authors.forename: Inna
authors.initials: I
authors.lastname: Goliand
authors.affiliation: Department of Life Sciences and the National Institute for Biotechnology in the Negev (NIBN), Ben-Gurion University of the Negev, Beer-Sheva 84105, Israel.
authors.forename: Dikla
authors.initials: D
authors.lastname: Nachmias
authors.affiliation: Department of Life Sciences and the National Institute for Biotechnology in the Negev (NIBN), Ben-Gurion University of the Negev, Beer-Sheva 84105, Israel.
authors.forename: Ofir
authors.initials: O
authors.lastname: Gershony
authors.affiliation: Department of Life Sciences and the National Institute for Biotechnology in the Negev (NIBN), Ben-Gurion University of the Negev, Beer-Sheva 84105, Israel.
authors.forename: Natalie
authors.initials: N
authors.lastname: Elia
mesh_terms: D000818:Animals; D016923:Cell Death / Q000235:genetics*; D048749:Cytokinesis / Q000235:genetics*; D004285:Dogs; D056827:Endosomal Sorting Complexes Required for Transport / Q000235:genetics / Q000378:metabolism*; D011992:Endosomes / Q000378:metabolism / Q000648:ultrastructure; D006801:Humans; D061985:Madin Darby Canine Kidney Cells; D008561:Membrane Fusion / Q000235:genetics; D011485:Protein Binding
chemical_list: C481528:CHMP6 protein, human; D056827:Endosomal Sorting Complexes Required for Transport