De-ubiquitination and ubiquitin ligase domains of A20 downregulate NF-kappaB signalling.
NF-kappaB transcription factors mediate the effects of pro-inflammatory cytokines such as tumour necrosis factor-alpha and interleukin-1beta. Failure to downregulate NF-kappaB transcriptional activity results in chronic inflammation and cell death, as observed in A20-deficient mice. A20 is a potent inhibitor of NF-kappaB signalling, but its mechanism of action is unknown. Here we show that A20 downregulates NF-kappaB signalling through the cooperative activity of its two ubiquitin-editing domains. The amino-terminal domain of A20, which is a de-ubiquitinating (DUB) enzyme of the OTU (ovarian tumour) family, removes lysine-63 (K63)-linked ubiquitin chains from receptor interacting protein (RIP), an essential mediator of the proximal TNF receptor 1 (TNFR1) signalling complex. The carboxy-terminal domain of A20, composed of seven C2/C2 zinc fingers, then functions as a ubiquitin ligase by polyubiquitinating RIP with K48-linked ubiquitin chains, thereby targeting RIP for proteasomal degradation. Here we define a novel ubiquitin ligase domain and identify two sequential mechanisms by which A20 downregulates NF-kappaB signalling. We also provide an example of a protein containing separate ubiquitin ligase and DUB domains, both of which participate in mediating a distinct regulatory effect.
Pubmed ID: 15258597 RIS Download
Cell Line | DNA-Binding Proteins | Down-Regulation | HeLa Cells | Humans | Intracellular Signaling Peptides and Proteins | NF-kappa B | Nuclear Proteins | Protein Structure, Tertiary | Proteins | RNA, Small Interfering | Receptor-Interacting Protein Serine-Threonine Kinases | Signal Transduction | TNF Receptor-Associated Factor 2 | Tumor Necrosis Factor alpha-Induced Protein 3 | Ubiquitin | Ubiquitin-Conjugating Enzymes | Ubiquitin-Protein Ligases