Abstract
The alternative or noncanonical NF-κB pathway regulates the generation of p52-containing NF-κB dimers (e.g., p52/RelB) through a partial degradation (called processing) of the precursor p100 into p52. This pathway is activated by a subset of non-death TNF receptor members, which ultimately activate two kinases: NIK (NF-κB-Inducing Kinase) and IKKα (Inhibitor of κB Kinase alpha). These kinases create a phosphodegron for the E3 ligase SCF-β-TrCP that covalently binds K48-linked polyubiquitin chain onto p100 prior to its proteasomal processing. The resulting p52-containing complexes translocate into the nucleus to activate target genes involved in secondary lymphoid organ development, B cell survival or in osteoclastogenesis.
We describe in this chapter straightforward methods to monitor the activation of the alternative NF-κB pathway. These methods uncover cytosolic and nuclear biochemical modifications of key proteins of the alternative NF-κB pathway required prior to the transcription of NF-κB target genes.
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Acknowledgments
We thank the Centre Anticancéreux (CAC) from the University of Liège (Belgium), the Fédération belge Contre le Cancer (FCC) (Belgium), and the Interuniversity Attraction Poles (IAP7/32) (Belgium) for their funding. C.R. is supported by a fellowship from the Télévie (Belgium), and E.D. is a Research Associate at the FNRS (Belgium).
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Remouchamps, C., Dejardin, E. (2015). Methods to Assess the Activation of the Alternative (Noncanonical) NF-κB Pathway by Non-death TNF Receptors. 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_7
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DOI: https://doi.org/10.1007/978-1-4939-2422-6_7
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