Elucidating dynamic protein–protein interactions and ubiquitination in NF-κB signaling pathways

Noula Shembade, Edward W. Harhaj

Research output: Contribution to journalArticle

3 Citations (Scopus)

Abstract

The Nuclear factor-kappaB (NF-κB) family of transcription factors plays critical roles in infl ammatory responses and host defense; however, uncontrolled NF-κB activation can be deleterious by promoting autoimmune diseases and cancers. Lysine K63 (K63)-linked polyubiquitination has emerged as an important regulatory mechanism in NF-κB signaling by regulating dynamic protein–protein interactions that trigger NF-κB signaling. RIP1 and TRAF6 serve as key substrates of K63-linked polyubiquitin chains in tumor necrosis factor receptor (TNFR) and interleukin-1 receptor (IL-1R) pathways respectively as a mechanism to recruit TAK1 and IKK kinases by associated ubiquitin-binding adaptor molecules. Activation of IKKβ by TAK1 induces IκBα phosphorylation, degradation, and downstream NF-κB activation. The ubiquitin-editing enzyme A20 maintains transient NF-κB activation by opposing the K63-linked polyubiquitination of RIP1 and TRAF6. A20 inducibly interacts with the adaptor molecule TAX1BP1 and the E3 ligases Itch and RNF11 to form an A20 ubiquitin-editing enzyme complex. Notably, loss-of-function somatic mutations or polymorphisms in human A20 are associated with B-cell lymphomas or a variety of autoimmune diseases as a result of dysregulated NF-κB activation. In this chapter, we summarize the protocols routinely used in our laboratories to examine ubiquitination and NF-κB signaling.

Original languageEnglish (US)
Pages (from-to)283-295
Number of pages13
JournalMethods in Molecular Biology
Volume1280
DOIs
StatePublished - Jan 1 2015

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Ubiquitination
Ubiquitin
TNF Receptor-Associated Factor 6
Autoimmune Diseases
Polyubiquitin
Ubiquitin-Protein Ligases
Interleukin-1 Receptors
Tumor Necrosis Factor Receptors
B-Cell Lymphoma
Enzymes
Lysine
Transcription Factors
Phosphotransferases
Phosphorylation
Mutation
Neoplasms

All Science Journal Classification (ASJC) codes

  • Molecular Biology
  • Genetics

Cite this

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abstract = "The Nuclear factor-kappaB (NF-κB) family of transcription factors plays critical roles in infl ammatory responses and host defense; however, uncontrolled NF-κB activation can be deleterious by promoting autoimmune diseases and cancers. Lysine K63 (K63)-linked polyubiquitination has emerged as an important regulatory mechanism in NF-κB signaling by regulating dynamic protein–protein interactions that trigger NF-κB signaling. RIP1 and TRAF6 serve as key substrates of K63-linked polyubiquitin chains in tumor necrosis factor receptor (TNFR) and interleukin-1 receptor (IL-1R) pathways respectively as a mechanism to recruit TAK1 and IKK kinases by associated ubiquitin-binding adaptor molecules. Activation of IKKβ by TAK1 induces IκBα phosphorylation, degradation, and downstream NF-κB activation. The ubiquitin-editing enzyme A20 maintains transient NF-κB activation by opposing the K63-linked polyubiquitination of RIP1 and TRAF6. A20 inducibly interacts with the adaptor molecule TAX1BP1 and the E3 ligases Itch and RNF11 to form an A20 ubiquitin-editing enzyme complex. Notably, loss-of-function somatic mutations or polymorphisms in human A20 are associated with B-cell lymphomas or a variety of autoimmune diseases as a result of dysregulated NF-κB activation. In this chapter, we summarize the protocols routinely used in our laboratories to examine ubiquitination and NF-κB signaling.",
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Elucidating dynamic protein–protein interactions and ubiquitination in NF-κB signaling pathways. / Shembade, Noula; Harhaj, Edward W.

In: Methods in Molecular Biology, Vol. 1280, 01.01.2015, p. 283-295.

Research output: Contribution to journalArticle

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