title: MLKL compromises plasma membrane integrity by binding to phosphatidylinositol phosphates.
abstract: Although mixed lineage kinase domain-like (MLKL) protein has emerged as a specific and crucial protein for necroptosis induction, how MLKL transduces the death signal remains poorly understood. Here, we demonstrate that the full four-helical bundle domain (4HBD) in the N-terminal region of MLKL is required and sufficient to induce its oligomerization and trigger cell death. Moreover, we found that a patch of positively charged amino acids on the surface of the 4HBD binds to phosphatidylinositol phosphates (PIPs) and allows recruitment of MLKL to the plasma membrane. Importantly, we found that recombinant MLKL, but not a mutant lacking these positive charges, induces leakage of PIP-containing liposomes as potently as BAX, supporting a model in which MLKL induces necroptosis by directly permeabilizing the plasma membrane. Accordingly, we found that inhibiting the formation of PI(5)P and PI(4,5)P2 specifically inhibits tumor necrosis factor (TNF)-mediated necroptosis but not apoptosis.
authors.affiliation: VIB Inflammation Research Center, Technologiepark 927, 9052 Zwijnaarde-Ghent, Belgium; Department of Biomedical Molecular Biology, Ghent University, Technologiepark 927, 9052 Zwijnaarde-Ghent, Belgium.
authors.forename: Yves
authors.initials: Y
authors.lastname: Dondelinger
authors.affiliation: VIB Inflammation Research Center, Technologiepark 927, 9052 Zwijnaarde-Ghent, Belgium; Department of Biomedical Molecular Biology, Ghent University, Technologiepark 927, 9052 Zwijnaarde-Ghent, Belgium.
authors.forename: Wim
authors.initials: W
authors.lastname: Declercq
authors.affiliation: Department of Cell Biology, University of Geneva, 1211 Geneva 4, Switzerland.
authors.forename: Sylvie
authors.initials: S
authors.lastname: Montessuit
authors.affiliation: VIB Inflammation Research Center, Technologiepark 927, 9052 Zwijnaarde-Ghent, Belgium; Department of Biomedical Molecular Biology, Ghent University, Technologiepark 927, 9052 Zwijnaarde-Ghent, Belgium.
authors.forename: Ria
authors.initials: R
authors.lastname: Roelandt
authors.affiliation: Microscopy Core Facility, VIB Inflammation Research Center, VIB/Ghent University, Technologiepark 927, 9052 Zwijnaarde-Ghent, Belgium.
authors.forename: Amanda
authors.initials: A
authors.lastname: Goncalves
authors.affiliation: VIB Inflammation Research Center, Technologiepark 927, 9052 Zwijnaarde-Ghent, Belgium; Department of Biomedical Molecular Biology, Ghent University, Technologiepark 927, 9052 Zwijnaarde-Ghent, Belgium.
authors.forename: Inge
authors.initials: I
authors.lastname: Bruggeman
authors.affiliation: VIB Inflammation Research Center, Technologiepark 927, 9052 Zwijnaarde-Ghent, Belgium; Department of Biomedical Molecular Biology, Ghent University, Technologiepark 927, 9052 Zwijnaarde-Ghent, Belgium.
authors.forename: Paco
authors.initials: P
authors.lastname: Hulpiau
authors.affiliation: VIB Inflammation Research Center, Technologiepark 927, 9052 Zwijnaarde-Ghent, Belgium; Department of Biomedical Molecular Biology, Ghent University, Technologiepark 927, 9052 Zwijnaarde-Ghent, Belgium.
authors.forename: Kathrin
authors.initials: K
authors.lastname: Weber
authors.affiliation: Pattern Recognition Receptor Discovery Performance Unit, Immuno-Inflammation Therapeutic Area, GlaxoSmithKline, Collegeville, PA 19426, USA.
authors.forename: Clark A
authors.initials: CA
authors.lastname: Sehon
authors.affiliation: Pattern Recognition Receptor Discovery Performance Unit, Immuno-Inflammation Therapeutic Area, GlaxoSmithKline, Collegeville, PA 19426, USA.
authors.forename: Robert W
authors.initials: RW
authors.lastname: Marquis
authors.affiliation: Pattern Recognition Receptor Discovery Performance Unit, Immuno-Inflammation Therapeutic Area, GlaxoSmithKline, Collegeville, PA 19426, USA.
authors.forename: John
authors.initials: J
authors.lastname: Bertin
authors.affiliation: Pattern Recognition Receptor Discovery Performance Unit, Immuno-Inflammation Therapeutic Area, GlaxoSmithKline, Collegeville, PA 19426, USA.
authors.forename: Peter J
authors.initials: PJ
authors.lastname: Gough
authors.affiliation: Unit for Structural Biology and Biophysics, Laboratory for Protein Biochemistry and Biomolecular Engineering, Ghent University, K.L. Ledeganckstraat 35, 9000 Ghent, Belgium.
authors.forename: Savvas
authors.initials: S
authors.lastname: Savvides
authors.affiliation: Department of Cell Biology, University of Geneva, 1211 Geneva 4, Switzerland.
authors.forename: Jean-Claude
authors.initials: JC
authors.lastname: Martinou
authors.affiliation: VIB Inflammation Research Center, Technologiepark 927, 9052 Zwijnaarde-Ghent, Belgium; Department of Biomedical Molecular Biology, Ghent University, Technologiepark 927, 9052 Zwijnaarde-Ghent, Belgium.
authors.forename: Mathieu J M
authors.initials: MJ
authors.lastname: Bertrand
authors.affiliation: VIB Inflammation Research Center, Technologiepark 927, 9052 Zwijnaarde-Ghent, Belgium; Department of Biomedical Molecular Biology, Ghent University, Technologiepark 927, 9052 Zwijnaarde-Ghent, Belgium; Methusalem Program, Ghent University, Technologiepark 927, 9052 Zwijnaarde-Ghent, Belgium. Electronic address: peter.vandenabeele@irc.vib-ugent.be.
authors.forename: Peter
authors.initials: P
authors.lastname: Vandenabeele
mesh_terms: D016923:Cell Death / Q000187:drug effects / Q000502:physiology; D002460:Cell Line; D002462:Cell Membrane / Q000201:enzymology / Q000378:metabolism; D057809:HEK293 Cells; D006801:Humans; D008081:Liposomes / Q000378:metabolism; D009336:Necrosis; D018129:Phosphatidylinositol Phosphates / Q000378:metabolism*; D010766:Phosphorylation; D011494:Protein Kinases / Q000378:metabolism* / Q000494:pharmacology; D011994:Recombinant Proteins / Q000494:pharmacology; D015398:Signal Transduction; D014409:Tumor Necrosis Factor-alpha / Q000037:antagonists & inhibitors / Q000494:pharmacology
chemical_list: D008081:Liposomes; D018129:Phosphatidylinositol Phosphates; D011994:Recombinant Proteins; D014409:Tumor Necrosis Factor-alpha; C570693:MLKL protein, human; D011494:Protein Kinases