Apoptosis inducing factor (AIF) mediates lethal redox stress induced by menadione

// Hesti Lina Wiraswati 1,2,3,4,5 , Emilie Hangen 1,2,3,4 , Ana Belén Sanz 1,2,3,4,6 , Ngoc-Vy Lam 1,2,3,4 , Camille Reinhardt 3,4,7 , Allan Sauvat 1,2,3,8 , Ariane Mogha 1,2,3,4 , Alberto Ortiz 6 , Guido Kroemer 1,2,3,8,9,10,11,12,* and Nazanine Modjtahedi 3,4,7,* 1 Equipe 11 labellisée Ligue Nationale contre le Cancer, Centre de Recherche des Cordeliers, Paris, France 2 INSERM, U1138, Paris, France 3 Gustave Roussy Cancer Campus, Villejuif, France 4 Faculty of Medicine, Université Paris-Saclay, Kremlin-Bicêtre, France 5 Institut Teknologi Bandung (ITB), Bandung, Indonesia 6 Laboratory of Nephrology, IIS-Fundacion Jimenez Diaz UAM and REDINREN, Madrid, Spain 7 INSERM, U1030, Villejuif, France 8 Metabolomics and Cell Biology Platforms, Gustave Roussy Cancer Campus, Villejuif, France 9 Université Paris Descartes, Sorbonne Paris Cité, Paris, France 10 Université Pierre et Marie Curie, Paris, France 11 Pôle de Biologie, Hôpital Européen Georges Pompidou, AP-HP, Paris, France 12 Department of Women’s and Children’s Health, Karolinska Institute, Karolinska University Hospital, Stockholm, Sweden * These authors share senior co-authorship Correspondence to: Nazanine Modjtahedi, email: // Guido Kroemer, email: // Keywords : mitochondria, quinone metabolization, oxidative stress, protein arylation, Autophagy Received : March 13, 2016 Accepted : September 30, 2016 Published : October 11, 2016 Abstract Mitochondrial apoptosis inducing factor (AIF) is a redox-active enzyme that participates to the biogenesis/maintenance of complex I of the respiratory chain, yet also contributes to catabolic reactions in the context of regulated cell death when AIF translocates to the cytosol and to the nucleus. Here we explore the contribution of AIF to cell death induced by menadione (2-methyl-1,4-naphtoquinone; also called vitamin K3) in conditions in which this pro-oxidant does not cause the mitochondrial release of AIF, yet causes caspase-independent cell killing. Depletion of AIF from human cancer cells reduced the cytotoxicity of menadione. This cytoprotective effect was accompanied by the maintenance of high levels of reduced glutathione (GSH), which are normally depleted by menadione. In addition, AIF depletion reduced the arylation of cellular proteins induced by menadione. This menadione-triggered arylation, which can be measured by a fluorescence assay, is completely suppressed by addition of exogenous glutathione or N-acetyl cysteine. Complex I inhibition by Rotenone did not mimic the cytoprotective action of AIF depletion. Altogether, these results are compatible with the hypothesis that mitochondrion-sessile AIF facilitates lethal redox cycling of menadione, thereby precipitating protein arylation and glutathione depletion.