title: Cardiolipin externalization to the outer mitochondrial membrane acts as an elimination signal for mitophagy in neuronal cells.
abstract: Recognition of injured mitochondria for degradation by macroautophagy is essential for cellular health, but the mechanisms remain poorly understood. Cardiolipin is an inner mitochondrial membrane phospholipid. We found that rotenone, staurosporine, 6-hydroxydopamine and other pro-mitophagy stimuli caused externalization of cardiolipin to the mitochondrial surface in primary cortical neurons and SH-SY5Y cells. RNAi knockdown of cardiolipin synthase or of phospholipid scramblase-3, which transports cardiolipin to the outer mitochondrial membrane, decreased the delivery of mitochondria to autophagosomes. Furthermore, we found that the autophagy protein microtubule-associated-protein-1 light chain 3 (LC3), which mediates both autophagosome formation and cargo recognition, contains cardiolipin-binding sites important for the engulfment of mitochondria by the autophagic system. Mutation of LC3 residues predicted as cardiolipin-interaction sites by computational modelling inhibited its participation in mitophagy. These data indicate that redistribution of cardiolipin serves as an 'eat-me' signal for the elimination of damaged mitochondria from neuronal cells.
authors.affiliation: Department of Pathology, University of Pittsburgh, Pittsburgh, PA 15213.
authors.forename: Charleen T
authors.initials: CT
authors.lastname: Chu
authors.affiliation: Department of Environmental and Occupational Health and Center for Free Radical and Antioxidant Health, University of Pittsburgh, Pittsburgh, PA 15213.|Department of Critical Care Medicine and Safar Center for Resuscitation Research, University of Pittsburgh, Pittsburgh, PA 15213.
authors.forename: Jing
authors.initials: J
authors.lastname: Ji
authors.affiliation: Department of Pathology, University of Pittsburgh, Pittsburgh, PA 15213.
authors.forename: Ruben K
authors.initials: RK
authors.lastname: Dagda
authors.affiliation: Department of Environmental and Occupational Health and Center for Free Radical and Antioxidant Health, University of Pittsburgh, Pittsburgh, PA 15213.
authors.forename: Jian Fei
authors.initials: JF
authors.lastname: Jiang
authors.affiliation: Department of Environmental and Occupational Health and Center for Free Radical and Antioxidant Health, University of Pittsburgh, Pittsburgh, PA 15213.
authors.forename: Yulia Y
authors.initials: YY
authors.lastname: Tyurina
authors.affiliation: Department of Environmental and Occupational Health and Center for Free Radical and Antioxidant Health, University of Pittsburgh, Pittsburgh, PA 15213.
authors.forename: Alexandr A
authors.initials: AA
authors.lastname: Kapralov
authors.affiliation: Department of Environmental and Occupational Health and Center for Free Radical and Antioxidant Health, University of Pittsburgh, Pittsburgh, PA 15213.
authors.forename: Vladimir A
authors.initials: VA
authors.lastname: Tyurin
authors.affiliation: Department of Structural Biology, University of Pittsburgh, Pittsburgh, PA 15213.
authors.forename: Naveena
authors.initials: N
authors.lastname: Yanamala
authors.affiliation: Department of Computational and Systems Biology, University of Pittsburgh, PA 15213.
authors.forename: Indira H
authors.initials: IH
authors.lastname: Shrivastava
authors.affiliation: Department of Structural Biology, University of Pittsburgh, Pittsburgh, PA 15213.
authors.forename: Dariush
authors.initials: D
authors.lastname: Mohammadyani
authors.affiliation: Department of Pathology, University of Pittsburgh, Pittsburgh, PA 15213.
authors.forename: Kent Zhi Qiang
authors.initials: KZQ
authors.lastname: Wang
authors.affiliation: Department of Pathology, University of Pittsburgh, Pittsburgh, PA 15213.
authors.forename: Jianhui
authors.initials: J
authors.lastname: Zhu
authors.affiliation: Department of Structural Biology, University of Pittsburgh, Pittsburgh, PA 15213.
authors.forename: Judith
authors.initials: J
authors.lastname: Klein-Seetharaman
authors.affiliation: Department of Environmental and Occupational Health and Center for Free Radical and Antioxidant Health, University of Pittsburgh, Pittsburgh, PA 15213.
authors.forename: Krishnakumar
authors.initials: K
authors.lastname: Balasubramanian
authors.affiliation: Department of Environmental and Occupational Health and Center for Free Radical and Antioxidant Health, University of Pittsburgh, Pittsburgh, PA 15213.
authors.forename: Andrew A
authors.initials: AA
authors.lastname: Amoscato
authors.affiliation: Department of Environmental and Occupational Health and Center for Free Radical and Antioxidant Health, University of Pittsburgh, Pittsburgh, PA 15213.
authors.forename: Grigory
authors.initials: G
authors.lastname: Borisenko
authors.affiliation: Department of Environmental and Occupational Health and Center for Free Radical and Antioxidant Health, University of Pittsburgh, Pittsburgh, PA 15213.
authors.forename: Zhentai
authors.initials: Z
authors.lastname: Huang
authors.affiliation: Department of Pathology, University of Pittsburgh, Pittsburgh, PA 15213.
authors.forename: Aaron M
authors.initials: AM
authors.lastname: Gusdon
authors.affiliation: Department of Environmental and Occupational Health and Center for Free Radical and Antioxidant Health, University of Pittsburgh, Pittsburgh, PA 15213.
authors.forename: Amin
authors.initials: A
authors.lastname: Cheikhi
authors.affiliation: Department of Pathology, University of Pittsburgh, Pittsburgh, PA 15213.
authors.forename: Erin K
authors.initials: EK
authors.lastname: Steer
authors.affiliation: Department of Pathology, University of Pittsburgh, Pittsburgh, PA 15213.
authors.forename: Ruth
authors.initials: R
authors.lastname: Wang
authors.affiliation: Department of Cell Biology and Physiology and Center for Biologic Imaging, University of Pittsburgh, Pittsburgh, PA 15213.
authors.forename: Catherine
authors.initials: C
authors.lastname: Baty
authors.affiliation: Department of Cell Biology and Physiology and Center for Biologic Imaging, University of Pittsburgh, Pittsburgh, PA 15213.
authors.forename: Simon
authors.initials: S
authors.lastname: Watkins
authors.affiliation: Department of Computational and Systems Biology, University of Pittsburgh, PA 15213.
authors.forename: Ivet
authors.initials: I
authors.lastname: Bahar
authors.affiliation: Department of Environmental and Occupational Health and Center for Free Radical and Antioxidant Health, University of Pittsburgh, Pittsburgh, PA 15213.|Department of Critical Care Medicine and Safar Center for Resuscitation Research, University of Pittsburgh, Pittsburgh, PA 15213.
authors.forename: Hülya
authors.initials: H
authors.lastname: Bayir
authors.affiliation: Department of Environmental and Occupational Health and Center for Free Radical and Antioxidant Health, University of Pittsburgh, Pittsburgh, PA 15213.
authors.forename: Valerian E
authors.initials: VE
authors.lastname: Kagan
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D001343:Autophagy / Q000187:drug effects; D001692:Biological Transport / Q000187:drug effects; D002308:Cardiolipins / Q000235:genetics / Q000378:metabolism*; D045744:Cell Line, Tumor; D002478:Cells, Cultured; D055785:Gene Knockdown Techniques; D006367:HeLa Cells; D006801:Humans; D008928:Mitochondria / Q000187:drug effects; D051336:Mitochondrial Membranes / Q000378:metabolism*; D063306:Mitophagy / Q000187:drug effects / Q000502:physiology*; D008958:Models, Molecular; D008969:Molecular Sequence Data; D009474:Neurons / Q000187:drug effects / Q000502:physiology*; D016627:Oxidopamine / Q000494:pharmacology; D017434:Protein Structure, Tertiary; D051381:Rats; D017207:Rats, Sprague-Dawley; D012402:Rotenone / Q000494:pharmacology; D015398:Signal Transduction; D014475:Uncoupling Agents / Q000494:pharmacology
chemical_list: D002308:Cardiolipins; D014475:Uncoupling Agents; D012402:Rotenone; D016627:Oxidopamine
references.citation: Mol Biol Cell. 2012 Jan;23(2):381-9
references.pmid: 22114351
references.citation: Nat Chem Biol. 2005 Sep;1(4):223-32
references.pmid: 16408039
references.citation: Biochemistry. 1996 Dec 10;35(49):15784-90
references.pmid: 8961941
references.citation: Free Radic Biol Med. 2008 Jun 1;44(11):1935-44
references.pmid: 18375209
references.citation: J Pathol. 2010 Jun;221(2):117-24
references.pmid: 20225337
references.citation: J Biol Chem. 2012 Jun 1;287(23):19094-104
references.pmid: 22505714
references.citation: Science. 2011 Jul 8;333(6039):228-33
references.pmid: 21617041
references.citation: J Biol Chem. 2006 Dec 8;281(49):37496-506
references.pmid: 16990283
references.citation: Neuron. 2011 Jun 23;70(6):1033-53
references.pmid: 21689593
references.citation: Diabetes. 2007 Mar;56(3):786-94
references.pmid: 17327449
references.citation: Nat Neurosci. 2010 Jul;13(7):805-11
references.pmid: 20581817
references.citation: Cell Death Dis. 2010;1:e18
references.pmid: 21364619
references.citation: Proc Natl Acad Sci U S A. 2010 Jan 5;107(1):378-83
references.pmid: 19966284
references.citation: Science. 2012 Nov 9;338(6108):815-8
references.pmid: 23042293
references.citation: Biochemistry. 2008 Apr 15;47(15):4518-29
references.pmid: 18358005
references.citation: J Cell Biol. 2010 Apr 19;189(2):211-21
references.pmid: 20404107
references.citation: EMBO J. 1999 Nov 15;18(22):6329-38
references.pmid: 10562545
references.citation: Nat Cell Biol. 2010 Dec;12(12):1154-65
references.pmid: 21102437
references.citation: J Biol Chem. 2006 Jun 30;281(26):17870-81
references.pmid: 16621793
references.citation: Curr Biol. 2012 Mar 20;22(6):545-52
references.pmid: 22342752
references.citation: Biochimie. 2002 Feb-Mar;84(2-3):143-52
references.pmid: 12022945
references.citation: J Cell Sci. 2008 Aug 15;121(Pt 16):2685-95
references.pmid: 18653543
references.citation: Nat Cell Biol. 2012 Jan 22;14(2):177-85
references.pmid: 22267086
references.citation: J Cell Biol. 2008 Dec 1;183(5):795-803
references.pmid: 19029340
references.citation: Nat Cell Biol. 2010 Feb;12(2):119-31
references.pmid: 20098416
references.citation: Autophagy. 2008 Aug;4(6):770-82
references.pmid: 18594198
references.citation: J Lipid Res. 2008 Aug;49(8):1607-20
references.pmid: 18077827
references.citation: Methods Enzymol. 2008;442:375-93
references.pmid: 18662580
references.citation: Brain Pathol. 2003 Oct;13(4):473-81
references.pmid: 14655753
references.citation: Antioxid Redox Signal. 2011 May 15;14(10):1919-28
references.pmid: 21126216
references.citation: J Biol Chem. 1990 Mar 25;265(9):4923-8
references.pmid: 2138622
references.citation: PLoS Biol. 2010 Jan 26;8(1):e1000298
references.pmid: 20126261
references.citation: Open Biol. 2012 May;2(5):120080
references.pmid: 22724072
references.citation: EMBO Rep. 2010 Jan;11(1):45-51
references.pmid: 20010802
references.citation: Genes Cells. 2004 Jul;9(7):611-8
references.pmid: 15265004
references.citation: Nature. 2007 Nov 15;450(7168):435-9
references.pmid: 17960135
references.citation: J Biol Chem. 2012 Nov 23;287(48):40652-60
references.pmid: 23060438
references.citation: J Biol Chem. 2009 May 15;284(20):13843-13855
references.pmid: 19279012
references.citation: Arch Biochem Biophys. 2003 May 1;413(1):41-52
references.pmid: 12706340
references.citation: Cell Res. 2012 Mar;22(3):473-89
references.pmid: 22310240
references.citation: Genes Cells. 2010 Aug;15(8):887-900
references.pmid: 20604804
references.citation: Autophagy. 2011 Mar;7(3):279-96
references.pmid: 21189453
references.citation: Dev Cell. 2009 Jul;17(1):98-109
references.pmid: 19619495
references.citation: Hum Mol Genet. 2011 Mar 1;20(5):927-40
references.pmid: 21147754
references.citation: J Biol Chem. 2013 Jan 4;288(1):111-21
references.pmid: 23150663
references.citation: J Exp Med. 2010 Aug 30;207(9):1807-17
references.pmid: 20805564
references.citation: Nat Neurosci. 2012 Oct;15(10):1407-13
references.pmid: 22922784