title: NDPK-D (NM23-H4)-mediated externalization of cardiolipin enables elimination of depolarized mitochondria by mitophagy.
abstract: Mitophagy is critical for cell homeostasis. Externalization of the inner mitochondrial membrane phospholipid, cardiolipin (CL), to the surface of the outer mitochondrial membrane (OMM) was identified as a mitophageal signal recognized by the microtubule-associated protein 1 light chain 3. However, the CL-translocating machinery remains unknown. Here we demonstrate that a hexameric intermembrane space protein, NDPK-D (or NM23-H4), binds CL and facilitates its redistribution to the OMM. We found that mitophagy induced by a protonophoric uncoupler, carbonyl cyanide m-chlorophenylhydrazone (CCCP), caused externalization of CL to the surface of mitochondria in murine lung epithelial MLE-12 cells and human cervical adenocarcinoma HeLa cells. RNAi knockdown of endogenous NDPK-D decreased CCCP-induced CL externalization and mitochondrial degradation. A R90D NDPK-D mutant that does not bind CL was inactive in promoting mitophagy. Similarly, rotenone and 6-hydroxydopamine triggered mitophagy in SH-SY5Y cells was also suppressed by knocking down of NDPK-D. In situ proximity ligation assay (PLA) showed that mitophagy-inducing CL-transfer activity of NDPK-D is closely associated with the dynamin-like GTPase OPA1, implicating fission-fusion dynamics in mitophagy regulation.
authors.affiliation: Department of Environmental and Occupational Health, Center for Free Radical and Antioxidant Health, University of Pittsburgh, Pittsburgh, PA, USA.
authors.forename: V E
authors.initials: VE
authors.lastname: Kagan
authors.affiliation: Department of Environmental and Occupational Health, Center for Free Radical and Antioxidant Health, University of Pittsburgh, Pittsburgh, PA, USA.
authors.forename: J
authors.initials: J
authors.lastname: Jiang
authors.affiliation: Department of Environmental and Occupational Health, Center for Free Radical and Antioxidant Health, University of Pittsburgh, Pittsburgh, PA, USA.
authors.forename: Z
authors.initials: Z
authors.lastname: Huang
authors.affiliation: Department of Environmental and Occupational Health, Center for Free Radical and Antioxidant Health, University of Pittsburgh, Pittsburgh, PA, USA.
authors.forename: Y Y
authors.initials: YY
authors.lastname: Tyurina
authors.affiliation: University Grenoble Alpes-UJF, Laboratory of Fundamental and Applied Bioenergetics (LBFA), and SFR Environmental and Systems Biology (BEeSy), U1055, Grenoble, France.|Inserm, U1055, Grenoble, France.
authors.forename: C
authors.initials: C
authors.lastname: Desbourdes
authors.affiliation: University Grenoble Alpes-UJF, Laboratory of Fundamental and Applied Bioenergetics (LBFA), and SFR Environmental and Systems Biology (BEeSy), U1055, Grenoble, France.|Inserm, U1055, Grenoble, France.
authors.forename: C
authors.initials: C
authors.lastname: Cottet-Rousselle
authors.affiliation: Department of Environmental and Occupational Health, Center for Free Radical and Antioxidant Health, University of Pittsburgh, Pittsburgh, PA, USA.
authors.forename: H H
authors.initials: HH
authors.lastname: Dar
authors.affiliation: Department of Pathology, University of Pittsburgh, Pittsburgh, PA, USA.
authors.forename: M
authors.initials: M
authors.lastname: Verma
authors.affiliation: Department of Environmental and Occupational Health, Center for Free Radical and Antioxidant Health, University of Pittsburgh, Pittsburgh, PA, USA.
authors.forename: V A
authors.initials: VA
authors.lastname: Tyurin
authors.affiliation: Department of Environmental and Occupational Health, Center for Free Radical and Antioxidant Health, University of Pittsburgh, Pittsburgh, PA, USA.
authors.forename: A A
authors.initials: AA
authors.lastname: Kapralov
authors.affiliation: Department of Environmental and Occupational Health, Center for Free Radical and Antioxidant Health, University of Pittsburgh, Pittsburgh, PA, USA.
authors.forename: A
authors.initials: A
authors.lastname: Cheikhi
authors.affiliation: Department of Environmental and Occupational Health, Center for Free Radical and Antioxidant Health, University of Pittsburgh, Pittsburgh, PA, USA.
authors.forename: G
authors.initials: G
authors.lastname: Mao
authors.affiliation: Department of Cell Biology and Physiology, University of Pittsburgh, Pittsburgh, PA, USA.
authors.forename: D
authors.initials: D
authors.lastname: Stolz
authors.affiliation: Department of Cell Biology and Physiology, University of Pittsburgh, Pittsburgh, PA, USA.
authors.forename: C M
authors.initials: CM
authors.lastname: St Croix
authors.affiliation: Department of Cell Biology and Physiology, University of Pittsburgh, Pittsburgh, PA, USA.
authors.forename: S
authors.initials: S
authors.lastname: Watkins
authors.affiliation: Department of Biological Sciences, Wayne State University, Detroit, MI, USA.
authors.forename: Z
authors.initials: Z
authors.lastname: Shen
authors.affiliation: Department of Biological Sciences, Wayne State University, Detroit, MI, USA.
authors.forename: Y
authors.initials: Y
authors.lastname: Li
authors.affiliation: Department of Biological Sciences, Wayne State University, Detroit, MI, USA.
authors.forename: M L
authors.initials: ML
authors.lastname: Greenberg
authors.affiliation: University Grenoble Alpes-UJF, Laboratory of Fundamental and Applied Bioenergetics (LBFA), and SFR Environmental and Systems Biology (BEeSy), U1055, Grenoble, France.|Inserm, U1055, Grenoble, France.
authors.forename: M
authors.initials: M
authors.lastname: Tokarska-Schlattner
authors.affiliation: Sorbonne Universités, UPMC Univ Paris 06, Saint-Antoine Research Center, INSERM UMR-S 938, Paris, France.|AP-HP, Service de Biochimie et Hormonologie, Hôpital Tenon, Paris, France.
authors.forename: M
authors.initials: M
authors.lastname: Boissan
authors.affiliation: Sorbonne Universités, UPMC Univ Paris 06, Saint-Antoine Research Center, INSERM UMR-S 938, Paris, France.
authors.forename: M-L
authors.initials: ML
authors.lastname: Lacombe
authors.affiliation: Department of Biochemistry and Biomedical Sciences, McMaster University, Hamilton, Ontario, Canada.
authors.forename: R M
authors.initials: RM
authors.lastname: Epand
authors.affiliation: Department of Pathology, University of Pittsburgh, Pittsburgh, PA, USA.
authors.forename: C T
authors.initials: CT
authors.lastname: Chu
authors.affiliation: Department of Medicine, Acute Lung Injury Center of Excellence, University of Pittsburgh, Pittsburgh, PA, USA.|Medical Specialty Service Line, Veterans Affairs Pittsburgh Healthcare System, Pittsburgh, PA, USA.
authors.forename: R K
authors.initials: RK
authors.lastname: Mallampalli
authors.affiliation: Department of Critical Care Medicine, University of Pittsburgh, Pittsburgh, PA, USA.
authors.forename: H
authors.initials: H
authors.lastname: Bayır
authors.affiliation: University Grenoble Alpes-UJF, Laboratory of Fundamental and Applied Bioenergetics (LBFA), and SFR Environmental and Systems Biology (BEeSy), U1055, Grenoble, France.|Inserm, U1055, Grenoble, France.
authors.forename: U
authors.initials: U
authors.lastname: Schlattner
mesh_terms: D000818:Animals; D001343:Autophagy / Q000187:drug effects; D002258:Carbonyl Cyanide m-Chlorophenyl Hydrazone / Q000633:toxicity; D002308:Cardiolipins / Q000032:analysis / Q000378:metabolism*; D002460:Cell Line; D020558:GTP Phosphohydrolases / Q000378:metabolism; D006367:HeLa Cells; D006801:Humans; D008247:Lysosomes / Q000378:metabolism / Q000473:pathology; D051379:Mice; D008869:Microtubule-Associated Proteins / Q000378:metabolism; D008928:Mitochondria / Q000378:metabolism* / Q000473:pathology; D051336:Mitochondrial Membranes / Q000378:metabolism*; D063306:Mitophagy / Q000187:drug effects; D016297:Mutagenesis, Site-Directed; D061107:Nucleoside Diphosphate Kinase D / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism*; D016627:Oxidopamine / Q000494:pharmacology; D011485:Protein Binding; D034622:RNA Interference; D012402:Rotenone / Q000494:pharmacology
chemical_list: D002308:Cardiolipins; D008869:Microtubule-Associated Proteins; D012402:Rotenone; D002258:Carbonyl Cyanide m-Chlorophenyl Hydrazone; D016627:Oxidopamine; C517946:NME4 protein, human; D061107:Nucleoside Diphosphate Kinase D; D020558:GTP Phosphohydrolases; C415198:OPA1 protein, human