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A Guide to Production, Crystallization, and Structure Determination of Human IKK1/α
Published on: November 2, 2018
Tumor necrosis factor-alpha induces RelA degradation via ubiquitination at lysine 195 to prevent excessive nuclear
Yihui Fan1, Renfang Mao, Yanling Zhao
1Department of Pediatrics, Texas Children's Cancer Center, Houston, TX 77030, USA.
Abstract:
Ubiquitination-mediated degradation of the RelA subunit of nuclear factor-kappaB (NF-kappaB) is critical for the termination of NF-kappaB activation. However, the precise mechanism for the ubiquitination of RelA is still not fully understood. Here we report that tumor necrosis factor-alpha (TNFalpha) induces RelA polyubiquitination at the lysine 195 residue, and this ubiquitination event is critical for the degradation of RelA and termination of TNFalpha-mediated NF-kappaB activation. Overexpression of a RelA mutant with an arginine substitution for the lysine 195 residue dramatically inhibits RelA polyubiquitination and induces a stronger NF-kappaB activation compared with the wild type. Reconstitution of RelA-deficient mouse embryo fibroblast cells with wild-type RelA or RelA containing a K195R mutation revealed the importance of this site in TNFalpha-mediated RelA polyubiquitination, degradation, and attenuation of NF-kappaB activation. Our finding is the first report that substitution of a key RelA lysine residue with arginine inhibits TNFalpha-induced RelA ubiquitination and enhances TNFalpha-induced NF-kappaB activation.
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