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NF-kappa B pp 355–370Cite as

Control of NF-κB Subunits by Ubiquitination

Part of the Methods in Molecular Biology book series (MIMB,volume 1280)

Abstract

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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  • DOI: 10.1007/978-1-4939-2422-6_21
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References

  1. Hershko A (1983) Ubiquitin: roles in protein modification and breakdown. Cell 34:11–12

    CrossRef  CAS  PubMed  Google Scholar 

  2. Komander D, Rape M (2012) The ubiquitin code. Annu Rev Biochem 81:203–229

    CrossRef  CAS  PubMed  Google Scholar 

  3. Pickart CM (2001) Mechanisms underlying ubiquitination. Annu Rev Biochem 70:503–533

    CrossRef  CAS  PubMed  Google Scholar 

  4. Bernassola F, Karin M, Ciechanover A, Melino G (2008) The HECT family of E3 ubiquitin ligases: multiple players in cancer development. Cancer Cell 14:10–21

    CrossRef  CAS  PubMed  Google Scholar 

  5. Petroski MD, Deshaies RJ (2005) Function and regulation of cullin-RING ubiquitin ligases. Nat Rev Mol Cell Biol 6:9–20

    CrossRef  CAS  PubMed  Google Scholar 

  6. Chen ZJ, Sun LJ (2009) Nonproteolytic functions of ubiquitin in cell signaling. Mol Cell 33:275–286

    CrossRef  CAS  PubMed  Google Scholar 

  7. Adhikari A, Chen ZJ (2009) Diversity of polyubiquitin chains. Dev Cell 16:485–486

    CrossRef  CAS  PubMed  Google Scholar 

  8. Chen J, Chen ZJ (2013) Regulation of NF-kappaB by ubiquitination. Curr Opin Immunol 25:4–12

    CrossRef  PubMed Central  CAS  PubMed  Google Scholar 

  9. Spencer E, Jiang J, Chen ZJ (1999) Signal-induced ubiquitination of IkappaBalpha by the F-box protein Slimb/beta-TrCP. Genes Dev 13:284–294

    CrossRef  PubMed Central  CAS  PubMed  Google Scholar 

  10. Lamothe B, Besse A, Campos AD, Webster WK, Wu H, Darnay BG (2007) Site-specific Lys-63-linked tumor necrosis factor receptor-associated factor 6 auto-ubiquitination is a critical determinant of I kappa B kinase activation. J Biol Chem 282:4102–4112

    CrossRef  PubMed Central  CAS  PubMed  Google Scholar 

  11. Ea CK, Deng L, Xia ZP, Pineda G, Chen ZJ (2006) Activation of IKK by TNFalpha requires site-specific ubiquitination of RIP1 and polyubiquitin binding by NEMO. Mol Cell 22:245–257

    CrossRef  CAS  PubMed  Google Scholar 

  12. Fong A, Sun SC (2002) Genetic evidence for the essential role of beta-transducin repeat-containing protein in the inducible processing of NF-kappa B2/p100. J Biol Chem 277:22111–22114

    CrossRef  CAS  PubMed  Google Scholar 

  13. Carmody RJ, Ruan Q, Palmer S, Hilliard B, Chen YH (2007) Negative regulation of toll-like receptor signaling by NF-kappaB p50 ubiquitination blockade. Science 317:675–678

    CrossRef  CAS  PubMed  Google Scholar 

  14. Saccani S, Marazzi I, Beg AA, Natoli G (2004) Degradation of promoter-bound p65/RelA is essential for the prompt termination of the nuclear factor kappaB response. J Exp Med 200:107–113

    CrossRef  PubMed Central  CAS  PubMed  Google Scholar 

  15. Geng H, Wittwer T, Dittrich-Breiholz O, Kracht M, Schmitz ML (2009) Phosphorylation of NF-kappaB p65 at Ser468 controls its COMMD1-dependent ubiquitination and target gene-specific proteasomal elimination. EMBO Rep 10:381–386

    CrossRef  PubMed Central  CAS  PubMed  Google Scholar 

  16. Ryo A, Suizu F, Yoshida Y, Perrem K, Liou YC, Wulf G, Rottapel R, Yamaoka S, Lu KP (2003) Regulation of NF-kappaB signaling by Pin1-dependent prolyl isomerization and ubiquitin-mediated proteolysis of p65/RelA. Mol Cell 12:1413–1426

    CrossRef  CAS  PubMed  Google Scholar 

  17. Maine GN, Mao X, Komarck CM, Burstein E (2007) COMMD1 promotes the ubiquitination of NF-kappaB subunits through a cullin-containing ubiquitin ligase. EMBO J 26:436–447

    CrossRef  PubMed Central  CAS  PubMed  Google Scholar 

  18. Mao X, Gluck N, Li D, Maine GN, Li H, Zaidi IW, Repaka A, Mayo MW, Burstein E (2009) GCN5 is a required cofactor for a ubiquitin ligase that targets NF-kappaB/RelA. Genes Dev 23:849–861

    CrossRef  PubMed Central  CAS  PubMed  Google Scholar 

  19. Li H, Wittwer T, Weber A, Schneider H, Moreno R, Maine GN, Kracht M, Schmitz ML, Burstein E (2011) Regulation of NF-kappaB activity by competition between RelA acetylation and ubiquitination. Oncogene 31: 611–623

    Google Scholar 

  20. Colleran A, Collins PE, O’Carroll C, Ahmed A, Mao X, McManus B, Kiely PA, Burstein E, Carmody RJ (2013) Deubiquitination of NF-kappaB by Ubiquitin-Specific Protease-7 promotes transcription. Proc Natl Acad Sci U S A 110:618–623

    CrossRef  PubMed Central  CAS  PubMed  Google Scholar 

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Correspondence to Ruaidhrí J. Carmody .

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Collins, P.E., Colleran, A., Carmody, R.J. (2015). Control of NF-κB Subunits by Ubiquitination. In: May, M. (eds) NF-kappa B. Methods in Molecular Biology, vol 1280. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-2422-6_21

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  • DOI: https://doi.org/10.1007/978-1-4939-2422-6_21

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  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-2421-9

  • Online ISBN: 978-1-4939-2422-6

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