Methodology to study NF-κB/RelA ubiquitination in vivo.
Nuclear factor-kappa B (NF-κB) is a family of transcription factors that regulate immune responses, cell proliferation, differentiation, and survival. Activity of the NF-κΒ pathway on a cellular level is tightly controlled through various mechanisms, one of which is the ubiquitin-dependent degradation of chromatin-bound NF-κB subunits. In general, the ubiquitination of NF-κB regulates the duration of gene transcription activated in response to inflammatory signals. In this article, we present protocols to examine the in vivo ubiquitination status of RelA, a critical protein of the NF-κB family.
Department of Internal Medicine, UT Southwestern Medical Center, Dallas, TX, 75235, USA.
Haiying
H
Li
Petro
P
Starokadomskyy
Ezra
E
Burstein
D015153:Blotting, Western / Q000379:methods; D002460:Cell Line; D015870:Gene Expression; D006801:Humans; D047468:Immunoprecipitation / Q000379:methods; D016328:NF-kappa B / Q000235:genetics / Q000378:metabolism*; D046988:Proteasome Endopeptidase Complex / Q000378:metabolism; D059748:Proteolysis; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism; D015398:Signal Transduction; D051996:Transcription Factor RelA / Q000235:genetics / Q000378:metabolism*; D025801:Ubiquitin / Q000235:genetics / Q000378:metabolism; D054875:Ubiquitination
D016328:NF-kappa B; D011993:Recombinant Fusion Proteins; D051996:Transcription Factor RelA; D025801:Ubiquitin; D046988:Proteasome Endopeptidase Complex