NF-kappa B pp 355-370 | Cite as

Control of NF-κB Subunits by Ubiquitination

  • Patricia E. Collins
  • Amy Colleran
  • Ruaidhrí J. CarmodyEmail author
Part of the Methods in Molecular Biology book series (MIMB, volume 1280)


NF-κB is an essential regulator of inflammation and is also required for normal immune development and homeostasis. The inducible activation of NF-κB by a wide range of immuno-receptors such as the toll-like receptors (TLR), Tumour Necrosis Factor receptor (TNFR), and antigen T cell and B cell receptors requires the ubiquitin-triggered proteasomal degradation of IκBα to promote the nuclear translocation and transcriptional activity of NF-κB dimers. More recently, an additional role for ubiquitination and proteasomal degradation in the control of NF-κB activity has been uncovered. In this case, it is the ubiquitination and proteasomal degradation of the NF-κB subunits that play a critical role in the termination of the NF-κB-dependent transcriptional response induced by receptor activation. The primary trigger of NF-κB ubiquitination is DNA binding by NF-κB dimers and is further controlled by specific phosphorylation events which regulate the interaction of NF-κB with the E3 ligase complex and the deubiquitinase enzyme USP7. It is the balance between ubiquitination and deubiquitination that shapes the NF-κB-mediated transcriptional response. This chapter describes methods for the analysis of NF-κB ubiquitination.

Key words

NF-κB Ubiquitin Immunoprecipitation Immunoblotting 


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Copyright information

© Springer Science+Business Media New York 2015

Authors and Affiliations

  • Patricia E. Collins
    • 1
  • Amy Colleran
    • 1
  • Ruaidhrí J. Carmody
    • 1
    Email author
  1. 1.Institute of Infection, Immunology and Inflammation, College of Medical, Veterinary and Life SciencesUniversity of GlasgowGlasgowUK

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