RIPK1 inhibits ZBP1-driven necroptosis during development

Nature. 2016 Dec 1;540(7631):129-133. doi: 10.1038/nature20559. Epub 2016 Nov 7.


Receptor-interacting protein kinase 1 (RIPK1) promotes cell survival-mice lacking RIPK1 die perinatally, exhibiting aberrant caspase-8-dependent apoptosis and mixed lineage kinase-like (MLKL)-dependent necroptosis. However, mice expressing catalytically inactive RIPK1 are viable, and an ill-defined pro-survival function for the RIPK1 scaffold has therefore been proposed. Here we show that the RIP homotypic interaction motif (RHIM) in RIPK1 prevents the RHIM-containing adaptor protein ZBP1 (Z-DNA binding protein 1; also known as DAI or DLM1) from activating RIPK3 upstream of MLKL. Ripk1RHIM/RHIM mice that expressed mutant RIPK1 with critical RHIM residues IQIG mutated to AAAA died around birth and exhibited RIPK3 autophosphorylation on Thr231 and Ser232, which is a hallmark of necroptosis, in the skin and thymus. Blocking necroptosis with catalytically inactive RIPK3(D161N), RHIM mutant RIPK3, RIPK3 deficiency, or MLKL deficiency prevented lethality in Ripk1RHIM/RHIM mice. Loss of ZBP1, which engages RIPK3 in response to certain viruses but previously had no defined role in development, also prevented perinatal lethality in Ripk1RHIM/RHIM mice. Consistent with the RHIM of RIPK1 functioning as a brake that prevents ZBP1 from engaging the RIPK3 RHIM, ZBP1 interacted with RIPK3 in Ripk1RHIM/RHIMMlkl-/- macrophages, but not in wild-type, Mlkl-/- or Ripk1RHIM/RHIMRipk3RHIM/RHIM macrophages. Collectively, these findings indicate that the RHIM of RIPK1 is critical for preventing ZBP1/RIPK3/MLKL-dependent necroptosis during development.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adaptor Proteins, Vesicular Transport / deficiency
  • Adaptor Proteins, Vesicular Transport / metabolism
  • Amino Acid Motifs
  • Animals
  • Animals, Newborn
  • Apoptosis*
  • Caspase 8 / genetics
  • Caspase 8 / metabolism
  • Embryo, Mammalian / cytology
  • Embryo, Mammalian / embryology*
  • Embryo, Mammalian / metabolism*
  • Female
  • Glycoproteins / antagonists & inhibitors*
  • Glycoproteins / chemistry
  • Glycoproteins / deficiency
  • Glycoproteins / metabolism*
  • Macrophages / metabolism
  • Male
  • Mice
  • Mutation
  • Necrosis*
  • Phosphorylation
  • Protein Binding
  • Protein Kinases / deficiency
  • Protein Kinases / metabolism
  • RNA-Binding Proteins
  • Receptor-Interacting Protein Serine-Threonine Kinases / chemistry
  • Receptor-Interacting Protein Serine-Threonine Kinases / deficiency
  • Receptor-Interacting Protein Serine-Threonine Kinases / genetics
  • Receptor-Interacting Protein Serine-Threonine Kinases / metabolism*
  • Tumor Necrosis Factor-alpha / pharmacology


  • Adaptor Proteins, Vesicular Transport
  • Glycoproteins
  • RNA-Binding Proteins
  • TICAM-1 protein, mouse
  • Tumor Necrosis Factor-alpha
  • Zbp1 protein, mouse
  • MLKL protein, mouse
  • Protein Kinases
  • Receptor-Interacting Protein Serine-Threonine Kinases
  • Ripk1 protein, mouse
  • Ripk3 protein, mouse
  • Caspase 8