title: Structural identification of the Vps18 β-propeller reveals a critical role in the HOPS complex stability and function.
abstract: Membrane fusion at the vacuole, the lysosome equivalent in yeast, requires the HOPS tethering complex, which is recruited by the Rab7 GTPase Ypt7. HOPS provides a template for the assembly of SNAREs and thus likely confers fusion at a distinct position on vacuoles. Five of the six subunits in HOPS have a similar domain prediction with strong similarity to COPII subunits and nuclear porins. Here, we show that Vps18 indeed has a seven-bladed β-propeller as its N-terminal domain by revealing its structure at 2.14 Å. The Vps18 N-terminal domain can interact with the N-terminal part of Vps11 and also binds to lipids. Although deletion of the Vps18 N-terminal domain does not preclude HOPS assembly, as revealed by negative stain electron microscopy, the complex is instable and cannot support membrane fusion in vitro. We thus conclude that the β-propeller of Vps18 is required for HOPS stability and function and that it can serve as a starting point for further structural analyses of the HOPS tethering complex.
authors.affiliation: From the Max-Delbrück Center for Molecular Medicine, Macromolecular Structure and Interaction Group, Robert-Rössle-Strasse 10, 13125 Berlin, Germany, Freie Universität Berlin, Chemistry and Biochemistry Institute, Takustrasse 6, 14195 Berlin, Germany.
authors.forename: Heide
authors.initials: H
authors.lastname: Behrmann
authors.affiliation: Department of Biology/Chemistry, Biochemistry Section, University of Osnabrück, Barbarastrasse 13, 49076 Osnabrück, Germany.
authors.forename: Anna
authors.initials: A
authors.lastname: Lürick
authors.affiliation: Department of Structural Biochemistry, Max-Planck Institute of Molecular Physiology, Otto-Hahn-Strasse 11, 44227 Dortmund, Germany, and.
authors.forename: Anne
authors.initials: A
authors.lastname: Kuhlee
authors.affiliation: Department of Biology/Chemistry, Biochemistry Section, University of Osnabrück, Barbarastrasse 13, 49076 Osnabrück, Germany.
authors.forename: Henning Kleine
authors.initials: HK
authors.lastname: Balderhaar
authors.affiliation: Department of Biology/Chemistry, Biochemistry Section, University of Osnabrück, Barbarastrasse 13, 49076 Osnabrück, Germany.
authors.forename: Cornelia
authors.initials: C
authors.lastname: Bröcker
authors.affiliation: Department of Biology/Chemistry, Structural Biology Section, University of Osnabrück, Barbarastrasse 13, 49076 Osnabrück, Germany.
authors.forename: Daniel
authors.initials: D
authors.lastname: Kümmel
authors.affiliation: Department of Biology/Chemistry, Biochemistry Section, University of Osnabrück, Barbarastrasse 13, 49076 Osnabrück, Germany.
authors.forename: Siegfried
authors.initials: S
authors.lastname: Engelbrecht-Vandré
authors.affiliation: From the Max-Delbrück Center for Molecular Medicine, Macromolecular Structure and Interaction Group, Robert-Rössle-Strasse 10, 13125 Berlin, Germany.
authors.forename: Ulrich
authors.initials: U
authors.lastname: Gohlke
authors.affiliation: Department of Structural Biochemistry, Max-Planck Institute of Molecular Physiology, Otto-Hahn-Strasse 11, 44227 Dortmund, Germany, and.
authors.forename: Stefan
authors.initials: S
authors.lastname: Raunser
authors.affiliation: From the Max-Delbrück Center for Molecular Medicine, Macromolecular Structure and Interaction Group, Robert-Rössle-Strasse 10, 13125 Berlin, Germany, Freie Universität Berlin, Chemistry and Biochemistry Institute, Takustrasse 6, 14195 Berlin, Germany, heinemann@mdc-berlin.de.
authors.forename: Udo
authors.initials: U
authors.lastname: Heinemann
authors.affiliation: Department of Biology/Chemistry, Biochemistry Section, University of Osnabrück, Barbarastrasse 13, 49076 Osnabrück, Germany, cu@uos.de.
authors.forename: Christian
authors.initials: C
authors.lastname: Ungermann
mesh_terms: D033942:Adaptor Proteins, Vesicular Transport / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D022181:COP-Coated Vesicles / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D046912:Multiprotein Complexes / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D055550:Protein Stability; D017433:Protein Structure, Secondary; D017434:Protein Structure, Tertiary; D012441:Saccharomyces cerevisiae / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D029701:Saccharomyces cerevisiae Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism
chemical_list: D033942:Adaptor Proteins, Vesicular Transport; D046912:Multiprotein Complexes; C071657:PEP3 protein, S cerevisiae; D029701:Saccharomyces cerevisiae Proteins
keywords: Crystal Structure; Endosome; HOPS; Membrane Fusion; Rab; SNARE Proteins; Tethering Complex; Vacuole