Requirement of FADD, NEMO, and BAX/BAK for aberrant mitochondrial function in tumor necrosis factor alpha-induced necrosis

Krishna M. Irrinki, Karthik Mallilankaraman, Roshan J. Thapa, Harish C. Chandramoorthy, Frank J. Smith, Neelakshi R. Jog, Rajesh Kumar Gandhirajan, Steven G. Kelsen, Steven R. Houser, Michael J. May, Siddharth Balachandran, Muniswamy Madesh

Research output: Contribution to journalArticlepeer-review

91 Scopus citations

Abstract

Necroptosis represents a form of alternative programmed cell death that is dependent on the kinase RIP1. RIP1-dependent necroptotic death manifests as increased reactive oxygen species (ROS) production in mitochondria and is accompanied by loss of ATP biogenesis and eventual dissipation of mitochondrial membrane potential. Here, we show that tumor necrosis factor alpha (TNF-α)-induced necroptosis requires the adaptor proteins FADD and NEMO. FADD was found to mediate formation of the TNF-α-induced pronecrotic RIP1-RIP3 kinase complex, whereas the IκB Kinase (IKK) subunit NEMO appears to function downstream of RIP1-RIP3. Interestingly, loss of RelA potentiated TNF-α-dependent necroptosis, indicating that NEMO regulates necroptosis independently of NF-κB. Using both pharmacologic and genetic approaches, we demonstrate that the overexpression of antioxidants alleviates ROS elevation and necroptosis. Finally, elimination of BAX and BAK or overexpression of Bcl-x L protects cells from necroptosis at a later step. These findings provide evidence that mitochondria play an amplifying role in inflammation-induced necroptosis.

Original languageEnglish
Pages (from-to)3745-3758
Number of pages14
JournalMolecular and Cellular Biology
Volume31
Issue number18
DOIs
StatePublished - Sep 2011

Keywords

  • Animals
  • Apoptosis/physiology
  • Blotting, Western
  • Fas-Associated Death Domain Protein/metabolism
  • Flow Cytometry
  • GTPase-Activating Proteins/metabolism
  • Gene Knockout Techniques
  • Immunoprecipitation
  • Intracellular Signaling Peptides and Proteins/metabolism
  • Membrane Potential, Mitochondrial
  • Mice
  • Mitochondria/metabolism
  • NF-kappa B/metabolism
  • Necrosis/metabolism
  • Reactive Oxygen Species
  • Receptor-Interacting Protein Serine-Threonine Kinases/metabolism
  • Signal Transduction
  • Transcription Factor RelA/antagonists & inhibitors
  • Tumor Necrosis Factor-alpha/metabolism
  • bcl-2 Homologous Antagonist-Killer Protein/deficiency
  • bcl-2-Associated X Protein/deficiency

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