E2 interaction and dimerization in the crystal structure of TRAF6

Nat Struct Mol Biol. 2009 Jun;16(6):658-66. doi: 10.1038/nsmb.1605. Epub 2009 May 24.

Abstract

Tumor necrosis factor (TNF) receptor-associated factor (TRAF)-6 mediates Lys63-linked polyubiquitination for NF-kappaB activation via its N-terminal RING and zinc finger domains. Here we report the crystal structures of TRAF6 and its complex with the ubiquitin-conjugating enzyme (E2) Ubc13. The RING and zinc fingers of TRAF6 assume a rigid, elongated structure. Interaction of TRAF6 with Ubc13 involves direct contacts of the RING and the preceding residues, and the first zinc finger has a structural role. Unexpectedly, this region of TRAF6 is dimeric both in the crystal and in solution, different from the trimeric C-terminal TRAF domain. Structure-based mutagenesis reveals that TRAF6 dimerization is crucial for polyubiquitin synthesis and autoubiquitination. Fluorescence resonance energy transfer analysis shows that TRAF6 dimerization induces higher-order oligomerization of full-length TRAF6. The mismatch of dimeric and trimeric symmetry may provide a mode of infinite oligomerization that facilitates ligand-dependent signal transduction of many immune receptors.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, N.I.H., Intramural
  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

  • Crystallography, X-Ray
  • Fluorescence Resonance Energy Transfer
  • Humans
  • Protein Conformation
  • Protein Multimerization
  • RING Finger Domains
  • TNF Receptor-Associated Factor 6 / chemistry*
  • Ubiquitin-Conjugating Enzymes / chemistry*
  • Ubiquitination
  • Zinc Fingers

Substances

  • TNF Receptor-Associated Factor 6
  • UBE2N protein, human
  • Ubiquitin-Conjugating Enzymes