Lipopolysaccharide-mediated interferon regulatory factor activation involves TBK1-IKKepsilon-dependent Lys(63)-linked polyubiquitination and phosphorylation of TANK/I-TRAF

J Biol Chem. 2007 Oct 26;282(43):31131-46. doi: 10.1074/jbc.M701690200. Epub 2007 Sep 6.

Abstract

Type I interferon gene induction relies on IKK-related kinase TBK1 and IKKepsilon-mediated phosphorylations of IRF3/7 through the Toll-like receptor-dependent signaling pathways. The scaffold proteins that assemble these kinase complexes are poorly characterized. We show here that TANK/ITRAF is required for the TBK1- and IKKepsilon-mediated IRF3/7 phosphorylations through some Toll-like receptor-dependent pathways and is part of a TRAF3-containing complex. Moreover, TANK is dispensable for the early phase of double-stranded RNA-mediated IRF3 phosphorylation. Interestingly, TANK is heavily phosphorylated by TBK1-IKKepsilon upon lipopolysaccharide stimulation and is also subject to lipopolysaccharide- and TBK1-IKKepsilon-mediated Lys(63)-linked polyubiquitination, a mechanism that does not require TBK1-IKKepsilon kinase activity. Thus, we have identified TANK as a scaffold protein that assembles some but not all IRF3/7-phosphorylating TBK1-IKKepsilon complexes and demonstrated that these kinases possess two functions, namely the phosphorylation of both IRF3/7 and TANK as well as the recruitment of an E3 ligase for Lys(63)-linked polyubiquitination of their scaffold protein, TANK.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adaptor Proteins, Signal Transducing / immunology*
  • Adaptor Proteins, Signal Transducing / metabolism
  • Animals
  • Cell Line
  • Enzyme Activation / immunology
  • Genes, Reporter
  • HeLa Cells
  • Humans
  • I-kappa B Kinase / immunology*
  • I-kappa B Kinase / metabolism
  • Interferon Regulatory Factors / immunology
  • Interferon Regulatory Factors / metabolism
  • Kidney / cytology
  • Lipopolysaccharides / immunology*
  • Lipopolysaccharides / metabolism
  • Luciferases / metabolism
  • Lysine / chemistry
  • Lysine / metabolism*
  • Macrophages / immunology
  • Macrophages / metabolism
  • Mice
  • Phosphorylation
  • Protein Serine-Threonine Kinases / immunology*
  • Protein Serine-Threonine Kinases / metabolism
  • RNA Interference
  • RNA, Small Interfering / metabolism
  • Transfection
  • Two-Hybrid System Techniques
  • Ubiquitination

Substances

  • Adaptor Proteins, Signal Transducing
  • Interferon Regulatory Factors
  • Lipopolysaccharides
  • RNA, Small Interfering
  • TANK protein, human
  • Luciferases
  • Protein Serine-Threonine Kinases
  • TBK1 protein, human
  • I-kappa B Kinase
  • Lysine