Lys63-linked polyubiquitination of IRAK-1 is required for interleukin-1 receptor- and toll-like receptor-mediated NF-kappaB activation

Mol Cell Biol. 2008 May;28(10):3538-47. doi: 10.1128/MCB.02098-07. Epub 2008 Mar 17.

Abstract

Stimulation through the interleukin-1 receptor (IL-1R) and some Toll-like receptors (TLRs) induces ubiquitination of TRAF6 and IRAK-1, signaling components required for NF-kappaB and mitogen-activated protein kinase activation. Here we show that although TRAF6 and IRAK-1 acquired Lys63 (K63)-linked polyubiquitin chains upon IL-1 stimulation, only ubiquitinated IRAK-1 bound NEMO, the regulatory subunit of IkappaB kinase (IKK). The sites of IRAK-1 ubiquitination were mapped to Lys134 and Lys180, and arginine substitution of these residues impaired IL-1R/TLR-mediated IRAK-1 ubiquitination, NEMO binding, and NF-kappaB activation. K63-linked ubiquitination of IRAK-1 required enzymatically active TRAF6, indicating that it is the physiologically relevant E3. Thus, K63-linked polyubiquitination of proximal signaling proteins is a common mechanism used by diverse innate immune receptors for recruiting IKK and activating NF-kappaB.

Publication types

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

MeSH terms

  • Animals
  • Base Sequence
  • Binding Sites
  • Cell Line
  • Cells, Cultured
  • DNA Primers / genetics
  • Humans
  • I-kappa B Kinase / chemistry
  • I-kappa B Kinase / genetics
  • I-kappa B Kinase / metabolism
  • Interleukin-1 Receptor-Associated Kinases / chemistry*
  • Interleukin-1 Receptor-Associated Kinases / genetics
  • Interleukin-1 Receptor-Associated Kinases / metabolism*
  • Lysine / chemistry
  • Mice
  • NF-kappa B / metabolism*
  • Receptors, Interleukin-1 / genetics
  • Receptors, Interleukin-1 / metabolism*
  • Recombinant Proteins / chemistry
  • Recombinant Proteins / genetics
  • Recombinant Proteins / metabolism
  • TNF Receptor-Associated Factor 6 / genetics
  • TNF Receptor-Associated Factor 6 / metabolism
  • Toll-Like Receptor 4 / genetics
  • Toll-Like Receptor 4 / metabolism
  • Toll-Like Receptors / genetics
  • Toll-Like Receptors / metabolism*
  • Transfection
  • Ubiquitination

Substances

  • DNA Primers
  • IKBKG protein, human
  • NF-kappa B
  • Receptors, Interleukin-1
  • Recombinant Proteins
  • TLR4 protein, human
  • TNF Receptor-Associated Factor 6
  • Toll-Like Receptor 4
  • Toll-Like Receptors
  • IRAK1 protein, human
  • Interleukin-1 Receptor-Associated Kinases
  • I-kappa B Kinase
  • Lysine