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PMID: 17980495 Published · ppublish English Journal Article Research Support, N.I.H., Extramural

Upregulation of BNIP3 and translocation to mitochondria mediates cyanide-induced apoptosis in cortical cells.

Neuroscience ·Vol. 150 ·No. 1 ·2007-11-30 ·Pages 159-67

Prabhakaran K, Li L, Zhang L, Borowitz JL, Isom GE

Abstract

Bcl-2/adenovirus E1B 19-kDa-interacting protein 3 (BNIP3), a Bcl-2 homology domain 3 (BH3) domain only protein, has been identified as a mitochondrial mediator of hypoxia-induced cell death. Since cyanide produces histotoxic anoxia (chemical hypoxia), the present study was undertaken in primary rat cortical cells to determine involvement of the BNIP3 signaling pathway in cyanide-induced death. Over a 20 h exposure KCN increased BNIP3 expression, followed by a concentration-related apoptotic death. To determine if BNIP3 plays a role in the cell death, expression was either increased with BNIP3 cDNA (BNIP3+) or knocked down with small interfering RNA (RNAi). In BNIP3+ cells, cyanide-induced apoptotic death was markedly enhanced and preceded by reduction of mitochondrial membrane potential (delta psim), release of cytochrome c from mitochondria and elevated caspase 3 and 7 activity. Pretreatment with the pan-caspase inhibitor N-benzyloxycarbonyl-Ala-Asp-fluoromethyl ketone (zVAD-fmk) suppressed BNIP3+-mediated cell death, thus confirming a caspase-dependent apoptosis. On the other hand, BNIP3 knockdown by RNAi or antagonism of BNIP3 by a transmembrane-deleted dominant-negative mutant (BNIP3 delta TM) markedly reduced cell death. Immunohistochemical imaging showed that cyanide stimulated translocation of BNIP3 from cytosol to mitochondria and displacement studies with BNIP3 delta TM showed that integration of BNIP3 into the mitochondrial outer membrane was necessary for the cell death. In BNIP3+ cells, cyclosporin-A, an inhibitor of mitochondrial pore transition, blocked the cyanide-induced reduction of delta psim and decreased the apoptotic death. These results demonstrate in cortical cells that cyanide induces a rapid upregulation of BNIP3 expression, followed by translocation to the mitochondrial outer membrane to reduce delta psim. This was followed by mitochondrial release of cytochrome c to execute a caspase-dependent cell death.

MeSH Terms
Analysis of Variance Animals Apoptosis/drug effects Caspases/metabolism Cells, Cultured Cerebral Cortex/cytology Cyanides/pharmacology Cyclosporine/pharmacology Dose-Response Relationship, Drug Drug Interactions Embryo, Mammalian In Situ Nick-End Labeling Membrane Potential, Mitochondrial/drug effects Membrane Proteins/genetics,metabolism Mitochondria/metabolism Mitochondrial Proteins Mutation/physiology Neurons/drug effects Protein Transport/drug effects,physiology Proto-Oncogene Proteins/genetics,metabolism RNA, Small Interfering/pharmacology Rats Time Factors Transfection/methods
Chemicals
BNIP3 protein, rat Cyanides Membrane Proteins Mitochondrial Proteins Proto-Oncogene Proteins RNA, Small Interfering Cyclosporine Caspases
Authors & Affiliations
5 authors, click to expand affiliations / ORCID
Prabhakaran K
Department of Medicinal Chemistry and Molecular Pharmacology, Purdue University, West Lafayette, IN 47907-1333, USA.
Li L
Zhang L
Borowitz J L
Isom G E
References (39)
39 references, click to expand
  1. HIF-1alpha and p53 promote hypoxia-induced delayed neuronal death in models of CNS ischemia.
    Exp Neurol. 1999 Sep;159(1):65-72 PMID: 10486175
  2. Cyanide-induced apoptosis involves oxidative-stress-activated NF-kappaB in cortical neurons.
    Toxicol Appl Pharmacol. 2000 Apr 15;164(2):196-205 PMID: 10764633
  3. Arsenic trioxide induces autophagic cell death in malignant glioma cells by upregulation of mitochondrial cell death protein BNIP3.
    Oncogene. 2005 Feb 3;24(6):980-91 PMID: 15592527
  4. p38 Mitogen-activated protein kinase regulates Bax translocation in cyanide-induced apoptosis.
    Toxicol Sci. 2003 Sep;75(1):99-107 PMID: 12805646
  5. Inducible expression of BNIP3 provokes mitochondrial defects and hypoxia-mediated cell death of ventricular myocytes.
    Circ Res. 2002 Aug 9;91(3):226-31 PMID: 12169648
  6. Hypoxia and acidosis activate cardiac myocyte death through the Bcl-2 family protein BNIP3.
    Proc Natl Acad Sci U S A. 2002 Oct 1;99(20):12825-30 PMID: 12226479
  7. Hypoxia induces apoptosis with enhanced expression of Fas antigen messenger RNA in cultured neonatal rat cardiomyocytes.
    Circ Res. 1994 Sep;75(3):426-33 PMID: 7520371
  8. Upregulation of BNIP3 promotes apoptosis of lung cancer cells that were induced by p53.
    Biochem Biophys Res Commun. 2006 Jul 28;346(2):501-7 PMID: 16765911
  9. Activation of Ras up-regulates pro-apoptotic BNIP3 in nitric oxide-induced cell death.
    J Biol Chem. 2006 Nov 10;281(45):33939-48 PMID: 16954213
  10. BNIP3 and genetic control of necrosis-like cell death through the mitochondrial permeability transition pore.
    Mol Cell Biol. 2000 Aug;20(15):5454-68 PMID: 10891486
  11. Nix and Nip3 form a subfamily of pro-apoptotic mitochondrial proteins.
    J Biol Chem. 1999 Jan 1;274(1):7-10 PMID: 9867803
  12. Dieldrin-induced oxidative stress and neurochemical changes contribute to apoptopic cell death in dopaminergic cells.
    Free Radic Biol Med. 2001 Dec 1;31(11):1473-85 PMID: 11728820
  13. A unique pathway of cardiac myocyte death caused by hypoxia-acidosis.
    J Exp Biol. 2004 Aug;207(Pt 18):3189-200 PMID: 15299040
  14. Nuclear localization of the hypoxia-regulated pro-apoptotic protein BNIP3 after global brain ischemia in the rat hippocampus.
    Brain Res. 2004 Mar 19;1001(1-2):133-42 PMID: 14972662
  15. Oxidative stress and cyclooxygenase-2 induction mediate cyanide-induced apoptosis of cortical cells.
    Toxicol Appl Pharmacol. 2002 Nov 15;185(1):55-63 PMID: 12460737
  16. NF-kappaB-mediated up-regulation of Bcl-X(S) and Bax contributes to cytochrome c release in cyanide-induced apoptosis.
    J Neurochem. 2002 May;81(4):842-52 PMID: 12065643
  17. The E1B 19K/Bcl-2-binding protein Nip3 is a dimeric mitochondrial protein that activates apoptosis.
    J Exp Med. 1997 Dec 15;186(12):1975-83 PMID: 9396766
  18. RNAi functions in cultured mammalian neurons.
    Proc Natl Acad Sci U S A. 2002 Sep 3;99(18):11926-9 PMID: 12192088
  19. Regulation of BNIP3 in normal and cancer cells.
    Mol Cells. 2006 Feb 28;21(1):1-6 PMID: 16511341
  20. Modulation of the NMDA receptor by cyanide: enhancement of receptor-mediated responses.
    J Pharmacol Exp Ther. 1997 Mar;280(3):1341-8 PMID: 9067322
  21. BH4 domain of antiapoptotic Bcl-2 family members closes voltage-dependent anion channel and inhibits apoptotic mitochondrial changes and cell death.
    Proc Natl Acad Sci U S A. 2000 Mar 28;97(7):3100-5 PMID: 10737788
  22. Cell-specific induction of apoptosis by microinjection of cytochrome c. Bcl-xL has activity independent of cytochrome c release.
    J Biol Chem. 1997 Nov 28;272(48):30299-305 PMID: 9374516
  23. Hypoxia, BNip3 proteins, and the mitochondrial death pathway in cardiomyocytes.
    Circ Res. 2002 Aug 9;91(3):183-5 PMID: 12169642
  24. Oxidative stress induces neuronal death by recruiting a protease and phosphatase-gated mechanism.
    J Biol Chem. 2001 Sep 14;276(37):35049-59 PMID: 11443132
  25. Bcl-2-related protein family gene expression during oligodendroglial differentiation.
    J Neurochem. 2003 Jun;85(6):1500-12 PMID: 12787069
  26. Cyanide enhancement of dopamine-induced apoptosis in mesencephalic cells involves mitochondrial dysfunction and oxidative stress.
    Neurotoxicology. 2003 Jun;24(3):333-42 PMID: 12782099
  27. Differential susceptibility of brain areas to cyanide involves different modes of cell death.
    Toxicol Appl Pharmacol. 1999 Apr 1;156(1):6-16 PMID: 10101094
  28. Hypoxia induces the expression of the pro-apoptotic gene BNIP3.
    Cell Death Differ. 2001 Apr;8(4):367-76 PMID: 11550088
  29. Co-transfection with cDNA encoding the Bcl family of anti-apoptotic proteins improves the efficiency of transfection in primary fetal neural stem cells.
    J Neurosci Methods. 2002 Jun 30;117(2):153-8 PMID: 12100980
  30. Nitric oxide induces BNIP3 expression that causes cell death in macrophages.
    Biochem Biophys Res Commun. 2004 Aug 20;321(2):298-305 PMID: 15358175
  31. BNips: a group of pro-apoptotic proteins in the Bcl-2 family.
    Apoptosis. 2003 Jun;8(3):229-36 PMID: 12766483
  32. Localization and developmental ontogeny of the pro-apoptotic Bnip3 mRNA in the postnatal rat cortex and hippocampus.
    Brain Res. 2006 Jul 19;1100(1):55-63 PMID: 16765336
  33. Up-regulation of uncoupling protein 2 by cyanide is linked with cytotoxicity in mesencephalic cells.
    J Pharmacol Exp Ther. 2005 Sep;314(3):1338-45 PMID: 15937145
  34. Regulated targeting of BAX to mitochondria.
    J Cell Biol. 1998 Oct 5;143(1):207-15 PMID: 9763432
  35. BNip3 and signal-specific programmed death in the heart.
    J Mol Cell Cardiol. 2005 Jan;38(1):35-45 PMID: 15623420
  36. BNIP3 heterodimerizes with Bcl-2/Bcl-X(L) and induces cell death independent of a Bcl-2 homology 3 (BH3) domain at both mitochondrial and nonmitochondrial sites.
    J Biol Chem. 2000 Jan 14;275(2):1439-48 PMID: 10625696
  37. Cyanide intoxication and its mechanism of antagonism.
    Annu Rev Pharmacol Toxicol. 1984;24:451-81 PMID: 6428300
  38. Production of reactive oxygen species by mitochondria: central role of complex III.
    J Biol Chem. 2003 Sep 19;278(38):36027-31 PMID: 12840017
  39. Cyanide induces different modes of death in cortical and mesencephalon cells.
    J Pharmacol Exp Ther. 2002 Nov;303(2):510-9 PMID: 12388630
Article Info
Journal
Neuroscience
Abbr.
Neuroscience
ISSN
0306-4522
DOI
10.1016/j.neuroscience.2007.07.033
Published
2007-11-30
Epub
2007-00-29
Pages
159-67
Language
English
Region
United States
NLM ID
7605074
PMCID
PMC2196214
Subset
IM
Grants
NIEHS NIH HHS · R01 ES004140 · United States
NIEHS NIH HHS · R01 ES004140-21 · United States
NIEHS NIH HHS · ES04140 · United States
External Links
PubMed source DOI: 10.1016/j.neuroscience.2007.07.033 PMC Full Text View journal details
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