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Tumor Necrosis Factor Receptor Associated Factors (TRAFs) 2 and 3 Form a Transcriptional Complex with Phosho-RNA

Jimmy El Hokayem1, George C Brittain2,3, Zafar Nawaz1

  • 1Department of Biochemistry and Molecular Biology, University of Miami, Miller School of Medicine, Miami, FL, USA.

Molecular Neurobiology
|February 5, 2016
PubMed

Insights

Tumor necrosis factor receptor-associated factors (TRAFs) are newly found in neuronal cell nuclei, with TRAF2 and TRAF3 entering upon CD40L stimulation. These TRAFs regulate gene transcription, impacting neuronal signaling pathways.

Area of Science:

  • Neuroscience
  • Molecular Biology
  • Immunology

Background:

  • Tumor necrosis factor receptor-associated factors (TRAFs) traditionally act as cytosolic adaptors in immune receptor signaling.
  • Their nuclear functions and roles in neuronal cells remain largely unexplored.

Purpose of the Study:

  • To investigate the subcellular localization and nuclear functions of TRAFs in neuronal cells.
  • To elucidate the role of TRAFs in CD40L-induced gene transcription in neurons.

Main Methods:

  • Immunofluorescence microscopy to detect TRAF localization in Neuro2a cells and primary cortical neurons.
  • Luciferase assays to assess the transcriptional regulatory activity of TRAFs.
  • Chromatin immunoprecipitation followed by quantitative PCR (ChIP-qPCR) to identify TRAFs in transcriptional complexes.
  • Western blotting to analyze H2b ubiquitination.

Main Results:

  • TRAFs are present in both the cytoplasm and nucleus of neuronal cells.
  • TRAF2 and TRAF3 translocate to the nucleus rapidly after CD40L stimulation.
  • TRAFs exhibit cell-specific transcriptional activation or repression.
  • TRAFs, p65, and RNA Polymerase II form a complex on the Icam-1 promoter.
  • TRAF2 nuclear import is essential for H2b ubiquitination, a transcription-permissive epigenetic mark.

Conclusions:

  • TRAFs possess nuclear functions in neuronal cells, extending beyond their known cytosolic roles.
  • TRAFs are integral components of CD40L-induced transcriptional complexes in neurons.
  • This study reveals a novel mechanism for epigenetic regulation of gene expression in neurons mediated by TRAFs.
Keywords:
CD40NF‐kB transcription factorNeuronNucleusTNF receptor associated factor (TRAF)TranscriptionTumor necrosis factor (TNF)

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