Caspase-8 scaffolding function and MLKL regulate NLRP3 inflammasome activation downstream of TLR3

Nat Commun. 2015 Jun 24:6:7515. doi: 10.1038/ncomms8515.

Abstract

TLR2 promotes NLRP3 inflammasome activation via an early MyD88-IRAK1-dependent pathway that provides a priming signal (signal 1) necessary for activation of the inflammasome by a second potassium-depleting signal (signal 2). Here we show that TLR3 binding to dsRNA promotes post-translational inflammasome activation through intermediate and late TRIF/RIPK1/FADD-dependent pathways. Both pathways require the scaffolding but not the catalytic function of caspase-8 or RIPK1. Only the late pathway requires kinase competent RIPK3 and MLKL function. Mechanistically, FADD/caspase-8 scaffolding function provides a post-translational signal 1 in the intermediate pathway, whereas in the late pathway it helps the oligomerization of RIPK3, which together with MLKL provides both signal 1 and 2 for inflammasome assembly. Cytoplasmic dsRNA activates NLRP3 independent of TRIF, RIPK1, RIPK3 or mitochondrial DRP1, but requires FADD/caspase-8 in wildtype macrophages to remove RIPK3 inhibition. Our study provides a comprehensive analysis of pathways that lead to NLRP3 inflammasome activation in response to dsRNA.

Publication types

  • Research Support, N.I.H., Extramural

MeSH terms

  • Adaptor Proteins, Vesicular Transport / genetics
  • Adaptor Proteins, Vesicular Transport / metabolism
  • Animals
  • Carrier Proteins / genetics
  • Carrier Proteins / metabolism*
  • Caspase 8 / genetics
  • Caspase 8 / metabolism*
  • Dynamins / genetics
  • Dynamins / metabolism
  • Fas-Associated Death Domain Protein / genetics
  • Fas-Associated Death Domain Protein / metabolism
  • Macrophages / metabolism*
  • Mice
  • Mice, Knockout
  • Mitochondria / metabolism
  • Myeloid Differentiation Factor 88 / genetics
  • Myeloid Differentiation Factor 88 / metabolism
  • NLR Family, Pyrin Domain-Containing 3 Protein
  • Protein Kinases / genetics
  • Protein Kinases / metabolism*
  • RNA, Double-Stranded / metabolism*
  • Receptor-Interacting Protein Serine-Threonine Kinases / genetics
  • Receptor-Interacting Protein Serine-Threonine Kinases / metabolism
  • Toll-Like Receptor 3 / metabolism*

Substances

  • Adaptor Proteins, Vesicular Transport
  • Carrier Proteins
  • Fadd protein, mouse
  • Fas-Associated Death Domain Protein
  • Myd88 protein, mouse
  • Myeloid Differentiation Factor 88
  • NLR Family, Pyrin Domain-Containing 3 Protein
  • Nlrp3 protein, mouse
  • RNA, Double-Stranded
  • TICAM-1 protein, mouse
  • TLR3 protein, mouse
  • Toll-Like Receptor 3
  • MLKL protein, mouse
  • Protein Kinases
  • Receptor-Interacting Protein Serine-Threonine Kinases
  • Ripk1 protein, mouse
  • Ripk3 protein, mouse
  • Casp8 protein, mouse
  • Caspase 8
  • Dnm1l protein, mouse
  • Dynamins