RIPK1 inhibits ZBP1-driven necroptosis during development.
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/RHIMmice 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/RHIMmice. 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/RHIMmice. 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/RHIMmacrophages. Collectively, these findings indicate that the RHIM of RIPK1 is critical for preventing ZBP1/RIPK3/MLKL-dependent necroptosis during development.
Department of Physiological Chemistry, Genentech, 1 DNA Way, South San Francisco, California 94080, USA.
Kim
K
Newton
Department of Physiological Chemistry, Genentech, 1 DNA Way, South San Francisco, California 94080, USA.
Katherine E
KE
Wickliffe
Department of Physiological Chemistry, Genentech, 1 DNA Way, South San Francisco, California 94080, USA.
Allie
A
Maltzman
Department of Physiological Chemistry, Genentech, 1 DNA Way, South San Francisco, California 94080, USA.
Debra L
DL
Dugger
Molecular Genetics of Cancer Division, The Walter and Eliza Hall Institute of Medical Research, 1G Royal Parade, Parkville, Victoria 3052, Australia.|Department of Medical Biology, Melbourne University, Victoria 3010, Australia.
Andreas
A
Strasser
Department of Proteomics and Biological Resources, Genentech, 1 DNA Way, South San Francisco, California 94080, USA.
Victoria C
VC
Pham
Department of Proteomics and Biological Resources, Genentech, 1 DNA Way, South San Francisco, California 94080, USA.
Jennie R
JR
Lill
Department of Molecular Biology, Genentech, 1 DNA Way, South San Francisco, California 94080, USA.
Merone
M
Roose-Girma
Department of Molecular Biology, Genentech, 1 DNA Way, South San Francisco, California 94080, USA.
Søren
S
Warming
Department of Pathology, Genentech, 1 DNA Way, South San Francisco, California 94080, USA.
Margaret
M
Solon
Department of Pathology, Genentech, 1 DNA Way, South San Francisco, California 94080, USA.
Hai
H
Ngu
Department of Pathology, Genentech, 1 DNA Way, South San Francisco, California 94080, USA.
Joshua D
JD
Webster
Department of Physiological Chemistry, Genentech, 1 DNA Way, South San Francisco, California 94080, USA.
Vishva M
VM
Dixit
D033942:Adaptor Proteins, Vesicular Transport / Q000172:deficiency / Q000378:metabolism; D020816:Amino Acid Motifs; D000818:Animals; D000831:Animals, Newborn; D017209:Apoptosis; D053181:Caspase 8 / Q000235:genetics / Q000378:metabolism; D004622:Embryo, Mammalian / Q000166:cytology / Q000196:embryology* / Q000378:metabolism*; D005260:Female; D006023:Glycoproteins / Q000037:antagonists & inhibitors* / Q000737:chemistry / Q000172:deficiency / Q000378:metabolism*; D008264:Macrophages / Q000378:metabolism; D008297:Male; D051379:Mice; D009154:Mutation; D009336:Necrosis; D010766:Phosphorylation; D011485:Protein Binding; D011494:Protein Kinases / Q000172:deficiency / Q000378:metabolism; D016601:RNA-Binding Proteins; D053422:Receptor-Interacting Protein Serine-Threonine Kinases / Q000737:chemistry / Q000172:deficiency / Q000235:genetics / Q000378:metabolism*; D014409:Tumor Necrosis Factor-alpha / Q000494:pharmacology
D033942:Adaptor Proteins, Vesicular Transport; D053181:Caspase 8; D006023:Glycoproteins; D011494:Protein Kinases; D016601:RNA-Binding Proteins; D053422:Receptor-Interacting Protein Serine-Threonine Kinases; D014409:Tumor Necrosis Factor-alpha; C476286:Ticam1 protein, mouse; C402632:Zbp1 protein, mouse; C570694:MLKL protein, mouse; C506251:Ripk1 protein, mouse; C485615:Ripk3 protein, mouse