A RNA Interference Screen Identifies RIP3 as an Essential Inducer of TNF-Induced Programmed Necrosis

  • YoungSik Cho
  • Sreerupa Challa
  • Francis Ka-Ming Chan
Conference paper
Part of the Advances in Experimental Medicine and Biology book series (AEMB, volume 691)


Recent evidence indicates that TNF-like death cytokines can induce apoptotic and non-apoptotic forms of cell death. We have coined the term “programmed necrosis” to describe caspase-independent cell death induced by TNF-like cytokines. Besides an obligate requirement for the protein serine/threonine kinase RIP1 and the production of reactive oxygen species (ROS), relatively little is known about the molecular mechanisms that control TNF-induced programmed necrosis. In order to further illuminate the molecular pathway that governs programmed necrosis, we performed a targeted RNA interference (RNAi) screen. Our screen identified RIP3, a RIP1 family member, as a specific mediator for programmed necrosis, but not apoptosis. Biochemical analyses show that assembly of the pro-necrotic RIP1–RIP3 complex critically regulates induction of programmed necrosis. The physiological relevance of RIP3-dependent programmed necrosis is demonstrated by the failure of RIP3-deficient mice to control vaccinia virus infections.



This work was supported by NIH grant AI065877. F.K.C. was a recipient of investigator awards from the Smith Family Foundation and the Cancer Research Institute.


  1. 1.
    Galluzzi L, Kepp O, Kroemer G(2009) RIP kinases initiate programmed necrosis. J Mol Cell Biol 2009 1(1):8–10Google Scholar
  2. 2.
    Kono H, Rock KL (2008) How dying cells alert the immune system to danger. Nat Rev Immunol 8(4):279–289CrossRefPubMedGoogle Scholar
  3. 3.
    Holler N et al (2000) Fas triggers an alternative, caspase-8-independent cell death pathway using the kinase RIP as effector molecule. Nat Immunol 1(6):489–495CrossRefPubMedGoogle Scholar
  4. 4.
    Chan FK et al (2003) A role for tumor necrosis factor receptor-2 and receptor-interacting protein in programmed necrosis and antiviral responses. J Biol Chem 278(51):51613–51621CrossRefPubMedGoogle Scholar
  5. 5.
    Lin Y et al (2004) Tumor necrosis factor-induced nonapoptotic cell death requires receptor-interacting protein-mediated cellular reactive oxygen species accumulation. J Biol Chem 279(11):10822–10828CrossRefPubMedGoogle Scholar
  6. 6.
    Cho YS et al (2009) Phosphorylation-driven assembly of the RIP1-RIP3 complex regulates programmed necrosis and virus-induced inflammation. Cell 137(6):1112–11123CrossRefPubMedGoogle Scholar
  7. 7.
    Micheau O, Tschopp J (2003) Induction of TNF receptor I-mediated apoptosis via two sequential signaling complexes. Cell 114(2):181–190CrossRefPubMedGoogle Scholar
  8. 8.
    Degterev A et al (2008) Identification of RIP1 kinase as a specific cellular target of necrostatins. Nat Chem Biol 4(5):313–321CrossRefPubMedGoogle Scholar
  9. 9.
    Vanlangenakker N et al (2008) Molecular mechanisms and pathophysiology of necrotic cell death. Curr Mol Med 8(3):207–220CrossRefPubMedGoogle Scholar
  10. 10.
    Kim YS et al (2007) TNF-induced activation of the Nox1 NADPH oxidase and its role in the induction of necrotic cell death. Mol Cell 26(5):675–687CrossRefPubMedGoogle Scholar
  11. 11.
    Yazdanpanah B et al (2009) Riboflavin kinase couples TNF receptor 1 to NADPH oxidase. Nature 460(7259):1159–1163CrossRefPubMedGoogle Scholar
  12. 12.
    Zhang J et al (1998) Fas-mediated apoptosis and activation-induced T-cell proliferation are defective in mice lacking FADD/Mort1. Nature 392(6673):296–300CrossRefPubMedGoogle Scholar
  13. 13.
    Chun HJ et al (2002) Pleiotropic defects in lymphocyte activation caused by caspase-8 mutations lead to human immunodeficiency. Nature 419(6905):395–399CrossRefPubMedGoogle Scholar
  14. 14.
    Zhang Y et al (2005) Conditional Fas-associated death domain protein (FADD): GFP knockout mice reveal FADD is dispensable in thymic development but essential in peripheral T cell homeostasis. J Immunol 175(5):3033–3044PubMedGoogle Scholar
  15. 15.
    Bell BD et al (2008) FADD and caspase-8 control the outcome of autophagic signaling in proliferating T cells. Proc Natl Acad Sci USA 105(43):16677–16682CrossRefPubMedGoogle Scholar
  16. 16.
    Ch’en IL et al (2008) Antigen-mediated T cell expansion regulated by parallel pathways of death. Proc Natl Acad Sci USA 105(45):17463–17468CrossRefPubMedGoogle Scholar
  17. 17.
    Li M, Beg AA (2000) Induction of necrotic-like cell death by tumor necrosis factor alpha and caspase inhibitors: novel mechanism for killing virus-infected cells. J Virol 74(16):7470–7477CrossRefPubMedGoogle Scholar
  18. 18.
    He S et al (2009) Receptor interacting protein kinase-3 determines cellular necrotic response to TNF-alpha. Cell 137(6):1100–1111CrossRefPubMedGoogle Scholar
  19. 19.
    Zhang DW et al (2009) RIP3, an energy metabolism regulator that switches TNF-induced cell death from apoptosis to necrosis. Science 325(5938):332–336CrossRefPubMedGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC 2011

Authors and Affiliations

  • YoungSik Cho
    • 1
    • 2
    • 3
  • Sreerupa Challa
    • 1
  • Francis Ka-Ming Chan
    • 1
  1. 1.Department of PathologyUniversity of Massachusetts Medical SchoolWorcesterUSA
  2. 2.Immunology and Virology ProgramUniversity of Massachusetts Medical SchoolWorcesterUSA
  3. 3.Center for Metabolic Syndrome Therapeutics, Bio-Organic Science DivisionKorea Research Institute of Chemical TechnologyDaejeonKorea

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