Intracellular localization and transcriptional regulation of tumor necrosis factor (TNF) receptor-associated factor 4 (TRAF4)

Eur J Biochem. 2002 Oct;269(19):4819-29. doi: 10.1046/j.1432-1033.2002.03180.x.


To gain insight in the subcellular localization of tumor necrosis factor receptor-associated factor (TRAF4) we analyzed GFP chimeras of full-length TRAF4 and various deletion mutants derived thereof. While TRAF4-GFP (T4-GFP) was clearly localized in the cytoplasm, the N-terminal deletion mutant, T4(259-470), comprising the TRAF domain of the molecule, and a C-terminal deletion mutant consisting mainly of the RING and zinc finger domains of TRAF4 were both localized predominantly to the nucleus. Passive nuclear localization of T4(259-470) can be ruled out as the TRAF domain of TRAF4 was sufficient to form high molecular weight complexes. T4(259-470) recruited full-length TRAF4 into the nucleus whereas TRAF4 was unable to change the nuclear localization of T4(259-470). Thus, it seems that individual T4(259-470) mutant molecules are sufficient to direct the respective TRAF4-T4(259-470) heteromeric complexes into the nucleus. In cells forming cell-cell contacts, TRAF4 was recruited to the sites of contact via its C-TRAF domain. The expression of some TRAF proteins is regulated by the NF-kappaB pathway. Thus, we investigated whether this pathway is also involved in the regulation of the TRAF4 gene. Indeed, in primary T-cells and Jurkat cells stimulated with the NF-kappaB inducers TNF or phorbol 12-myristate 13-acetate (PMA), TRAF4-mRNA was rapidly up-regulated. In Jurkat T-cells deficient for I-kappaB kinase gamma (IKKgamma, also known as NEMO), an essential component of the NF-kappaB-inducing-IKK complex, induction of TRAF4 was completely inhibited. In cells deficient for RIP (receptor interactive protein), an essential signaling intermediate of TNF-dependent NF-kappaB activation, TNF-, but not PMA-induced up-regulation of TRAF4 was blocked. These data suggest that activation of the NF-kappaB pathway is involved in up-regulation of TRAF4 in T-cells.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Cell Adhesion
  • Cell Line
  • Cell Nucleus / metabolism
  • Cytoplasm / metabolism
  • Green Fluorescent Proteins
  • HeLa Cells
  • Humans
  • I-kappa B Kinase
  • In Vitro Techniques
  • Jurkat Cells
  • Luminescent Proteins / chemistry
  • Luminescent Proteins / genetics
  • Luminescent Proteins / metabolism
  • Lymphocyte Activation
  • Microscopy, Confocal
  • Microscopy, Fluorescence
  • NF-kappa B / metabolism
  • Protein Structure, Tertiary
  • Protein-Serine-Threonine Kinases / metabolism
  • Proteins / chemistry
  • Proteins / genetics*
  • Proteins / metabolism*
  • Receptors, Tumor Necrosis Factor / metabolism
  • Recombinant Fusion Proteins / chemistry
  • Recombinant Fusion Proteins / genetics
  • Recombinant Fusion Proteins / metabolism
  • Sequence Deletion
  • T-Lymphocytes / immunology
  • T-Lymphocytes / metabolism
  • TNF Receptor-Associated Factor 4
  • Tumor Necrosis Factor Receptor-Associated Peptides and Proteins
  • Up-Regulation


  • Luminescent Proteins
  • NF-kappa B
  • Proteins
  • Receptors, Tumor Necrosis Factor
  • Recombinant Fusion Proteins
  • TNF Receptor-Associated Factor 4
  • TRAF4 protein, human
  • Tumor Necrosis Factor Receptor-Associated Peptides and Proteins
  • Green Fluorescent Proteins
  • Protein-Serine-Threonine Kinases
  • CHUK protein, human
  • I-kappa B Kinase
  • IKBKB protein, human
  • IKBKE protein, human