title: Quantitative interaction mapping reveals an extended UBX domain in ASPL that disrupts functional p97 hexamers.
abstract: Interaction mapping is a powerful strategy to elucidate the biological function of protein assemblies and their regulators. Here, we report the generation of a quantitative interaction network, directly linking 14 human proteins to the AAA+ ATPase p97, an essential hexameric protein with multiple cellular functions. We show that the high-affinity interacting protein ASPL efficiently promotes p97 hexamer disassembly, resulting in the formation of stable p97:ASPL heterotetramers. High-resolution structural and biochemical studies indicate that an extended UBX domain (eUBX) in ASPL is critical for p97 hexamer disassembly and facilitates the assembly of p97:ASPL heterotetramers. This spontaneous process is accompanied by a reorientation of the D2 ATPase domain in p97 and a loss of its activity. Finally, we demonstrate that overproduction of ASPL disrupts p97 hexamer function in ERAD and that engineered eUBX polypeptides can induce cell death, providing a rationale for developing anti-cancer polypeptide inhibitors that may target p97 activity.
authors.affiliation: Max Delbrück Center for Molecular Medicine, Robert-Rössle-Straße 10, 13125 Berlin, Germany.
authors.forename: Anup
authors.initials: A
authors.lastname: Arumughan
authors.affiliation: Max Delbrück Center for Molecular Medicine, Robert-Rössle-Straße 10, 13125 Berlin, Germany.
authors.forename: Yvette
authors.initials: Y
authors.lastname: Roske
authors.affiliation: Max Delbrück Center for Molecular Medicine, Robert-Rössle-Straße 10, 13125 Berlin, Germany.
authors.forename: Carolin
authors.initials: C
authors.lastname: Barth
authors.affiliation: Max Delbrück Center for Molecular Medicine, Robert-Rössle-Straße 10, 13125 Berlin, Germany.
authors.forename: Laura Lleras
authors.initials: LL
authors.lastname: Forero
authors.affiliation: Max-Planck-Institute for Coal Research, Kaiser-Wilhelm-Platz 1, 45470 Mülheim an der Ruhr, Germany.
authors.forename: Kenny
authors.initials: K
authors.lastname: Bravo-Rodriguez
authors.affiliation: Max Delbrück Center for Molecular Medicine, Robert-Rössle-Straße 10, 13125 Berlin, Germany.
authors.forename: Alexandra
authors.initials: A
authors.lastname: Redel
authors.affiliation: Max Delbrück Center for Molecular Medicine, Robert-Rössle-Straße 10, 13125 Berlin, Germany.
authors.forename: Simona
authors.initials: S
authors.lastname: Kostova
authors.affiliation: Max Delbrück Center for Molecular Medicine, Robert-Rössle-Straße 10, 13125 Berlin, Germany.
authors.forename: Erik
authors.initials: E
authors.lastname: McShane
authors.affiliation: Max Delbrück Center for Molecular Medicine, Robert-Rössle-Straße 10, 13125 Berlin, Germany.
authors.forename: Robert
authors.initials: R
authors.lastname: Opitz
authors.affiliation: Max Delbrück Center for Molecular Medicine, Robert-Rössle-Straße 10, 13125 Berlin, Germany.
authors.forename: Katja
authors.initials: K
authors.lastname: Faelber
authors.affiliation: Max Delbrück Center for Molecular Medicine, Robert-Rössle-Straße 10, 13125 Berlin, Germany.
authors.forename: Kirstin
authors.initials: K
authors.lastname: Rau
authors.affiliation: Max Planck Institute for Molecular Genetics, Ihnestraße 63-73, 14194 Berlin, Germany.
authors.forename: Thorsten
authors.identifier: 0000-0001-9275-3146
authors.initials: T
authors.lastname: Mielke
authors.affiliation: Max Delbrück Center for Molecular Medicine, Robert-Rössle-Straße 10, 13125 Berlin, Germany.
authors.forename: Oliver
authors.initials: O
authors.lastname: Daumke
authors.affiliation: Max Delbrück Center for Molecular Medicine, Robert-Rössle-Straße 10, 13125 Berlin, Germany.
authors.forename: Matthias
authors.initials: M
authors.lastname: Selbach
authors.affiliation: Max-Planck-Institute for Coal Research, Kaiser-Wilhelm-Platz 1, 45470 Mülheim an der Ruhr, Germany.
authors.forename: Elsa
authors.initials: E
authors.lastname: Sanchez-Garcia
authors.affiliation: Max Delbrück Center for Molecular Medicine, Robert-Rössle-Straße 10, 13125 Berlin, Germany.
authors.forename: Oliver
authors.identifier: 0000-0001-6349-9193
authors.initials: O
authors.lastname: Rocks
authors.affiliation: Max Delbrück Center for Molecular Medicine, Robert-Rössle-Straße 10, 13125 Berlin, Germany.
authors.forename: Daniela
authors.initials: D
authors.lastname: Panáková
authors.affiliation: Max Delbrück Center for Molecular Medicine, Robert-Rössle-Straße 10, 13125 Berlin, Germany.|Institute for Chemistry and Biochemistry, Freie Universität Berlin, Takustraße 6, 14195 Berlin, Germany.
authors.forename: Udo
authors.initials: U
authors.lastname: Heinemann
authors.affiliation: Max Delbrück Center for Molecular Medicine, Robert-Rössle-Straße 10, 13125 Berlin, Germany.
authors.forename: Erich E
authors.initials: EE
authors.lastname: Wanker
mesh_terms: D001921:Brain / Q000473:pathology; D049109:Cell Proliferation; D018360:Crystallography, X-Ray; D060746:Endoplasmic Reticulum-Associated Degradation / Q000502:physiology*; D057809:HEK293 Cells; D006801:Humans; D047908:Intracellular Signaling Peptides and Proteins; D009154:Mutation; D015514:Oncogene Proteins, Fusion / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism*; D010455:Peptides / Q000235:genetics / Q000378:metabolism; D011485:Protein Binding; D000072417:Protein Domains / Q000502:physiology*; D015202:Protein Engineering; D060066:Protein Interaction Maps; D055503:Protein Multimerization; D020836:Protein Structure, Quaternary; D011994:Recombinant Proteins / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism; D000074405:Valosin Containing Protein / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism*
chemical_list: C422743:ASPSCR1 protein, human; D047908:Intracellular Signaling Peptides and Proteins; D015514:Oncogene Proteins, Fusion; D010455:Peptides; D011994:Recombinant Proteins; C000614178:VCP protein, human; D000074405:Valosin Containing Protein