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 language | English (US) |
|---|---|
| Pages (from-to) | 283-295 |
| Number of pages | 13 |
| Journal | Methods in Molecular Biology |
| Volume | 1280 |
| DOIs | |
| State | Published - Jan 1 2015 |
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All Science Journal Classification (ASJC) codes
- Molecular Biology
- Genetics
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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 journal › Article
TY - JOUR
T1 - Elucidating dynamic protein–protein interactions and ubiquitination in NF-κB signaling pathways
AU - Shembade, Noula
AU - Harhaj, Edward W.
PY - 2015/1/1
Y1 - 2015/1/1
N2 - 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.
AB - 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.
UR - http://www.scopus.com/inward/record.url?scp=84924362849&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=84924362849&partnerID=8YFLogxK
U2 - 10.1007/978-1-4939-2422-6_16
DO - 10.1007/978-1-4939-2422-6_16
M3 - Article
C2 - 25736755
AN - SCOPUS:84924362849
VL - 1280
SP - 283
EP - 295
JO - Methods in Molecular Biology
JF - Methods in Molecular Biology
SN - 1064-3745
ER -