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CD40 regulates the processing of NF-kappaB2 p100 to p52.

H J Coope1, P G P Atkinson, B Huhse

  • 1Divisions of Immune Cell Biology and Yeast Genetics, National Institute for Medical Research, London NW7 1AA, UK.

The EMBO Journal
|October 11, 2002
PubMed
Summary

CD40 ligation triggers the processing of NF-kappaB inhibitory protein p100 into the active p52 subunit, a crucial step in adaptive immunity. This pathway requires NF-kappaB-inducing kinase (NIK) for p100 processing.

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Area of Science:

  • Immunology
  • Molecular Biology
  • Cell Signaling

Background:

  • The nf-kb2 gene product, p100, is a precursor to the active p52 NF-kappaB subunit.
  • The specific ligands that induce p100 processing remain largely undefined.

Purpose of the Study:

  • To identify ligands that stimulate p100 processing.
  • To elucidate the mechanism by which CD40 ligation affects p100 processing and NF-kappaB activation.

Main Methods:

  • Transfection of 293 cells with CD40.
  • Analysis of p100 ubiquitylation and proteasome-mediated proteolysis.
  • Assessment of p52 nuclear translocation.
  • Studies on primary murine splenic B cells.
  • Investigation of NF-kappaB-inducing kinase (NIK) involvement.

Main Results:

  • CD40 ligation on 293 cells stimulates p100 ubiquitylation and subsequent proteasome-mediated proteolysis, leading to p52 production.
  • CD40-mediated p52 accumulation requires de novo protein synthesis and results in nuclear translocation, forming active NF-kappaB dimers.
  • Endogenous CD40 ligation on splenic B cells also induces p100 processing and delayed p52-RelB dimer nuclear translocation.
  • CD40-induced p100 processing is dependent on functional NF-kappaB-inducing kinase (NIK) in both cell types.
  • NIK activity is not required for CD40-mediated IkappaBalpha degradation.

Conclusions:

  • CD40 ligation is a novel trigger for p100 processing to p52.
  • The CD40-NIK pathway regulates p52 generation, impacting NF-kappaB activity.
  • This mechanism is likely critical for transcriptional regulation of CD40 target genes in adaptive immunity.