TRAF Family Member-associated NF-κB Activator (TANK) Inhibits Genotoxic Nuclear Factor κB Activation by Facilitating Deubiquitinase USP10-dependent Deubiquitination of TRAF6 Ligase.
DNA damage-induced NF-κB activation plays a critical role in regulating cellular response to genotoxic stress. However, the molecular mechanisms controlling the magnitude and duration of this genotoxic NF-κB signaling cascade are poorly understood. We recently demonstrated that genotoxic NF-κB activation is regulated by reversible ubiquitination of several essential mediators involved ... in this signaling pathway. Here we show that TRAF family member-associated NF-κB activator (TANK) negatively regulates NF-κB activation by DNA damage via inhibiting ubiquitination of TRAF6. Despite the lack of a deubiquitination enzyme domain, TANK has been shown to negatively regulate the ubiquitination of TRAF proteins. We found TANK formed a complex with MCPIP1 (also known as ZC3H12A) and a deubiquitinase, USP10, which was essential for the USP10-dependent deubiquitination of TRAF6 and the resolution of genotoxic NF-κB activation upon DNA damage. Clustered regularly interspaced short palindromic repeats (CRISPR)/Cas9-mediated deletion of TANK in human cells significantly enhanced NF-κB activation by genotoxic treatment, resulting in enhanced cell survival and increased inflammatory cytokine production. Furthermore, we found that the TANK-MCPIP1-USP10 complex also decreased TRAF6 ubiquitination in cells treated with IL-1β or LPS. In accordance, depletion of USP10 enhanced NF-κB activation induced by IL-1β or LPS. Collectively, our data demonstrate that TANK serves as an important negative regulator of NF-κB signaling cascades induced by genotoxic stress and IL-1R/Toll-like receptor stimulation in a manner dependent on MCPIP1/USP10-mediated TRAF6 deubiquitination.
Mesh Terms:
Adaptor Proteins, Signal Transducing, Animals, Apoptosis, Blotting, Western, Cell Proliferation, Cells, Cultured, DNA Damage, Gene Expression Regulation, Humans, Immunoprecipitation, Mice, Mice, Knockout, NF-kappa B, Protein Processing, Post-Translational, RNA, Messenger, Real-Time Polymerase Chain Reaction, Receptors, Interleukin-1, Reverse Transcriptase Polymerase Chain Reaction, Ribonucleases, Signal Transduction, TNF Receptor-Associated Factor 6, Toll-Like Receptors, Ubiquitin, Ubiquitin Thiolesterase, Ubiquitination
Adaptor Proteins, Signal Transducing, Animals, Apoptosis, Blotting, Western, Cell Proliferation, Cells, Cultured, DNA Damage, Gene Expression Regulation, Humans, Immunoprecipitation, Mice, Mice, Knockout, NF-kappa B, Protein Processing, Post-Translational, RNA, Messenger, Real-Time Polymerase Chain Reaction, Receptors, Interleukin-1, Reverse Transcriptase Polymerase Chain Reaction, Ribonucleases, Signal Transduction, TNF Receptor-Associated Factor 6, Toll-Like Receptors, Ubiquitin, Ubiquitin Thiolesterase, Ubiquitination
J. Biol. Chem.
Date: May. 22, 2015
PubMed ID: 25861989
View in: Pubmed Google Scholar
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