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A20/Nrdp1 interaction alters the inflammatory signaling profile by mediating K48- and K63-linked polyubiquitination of effectors MyD88 and TBK1.
- Source :
-
The Journal of biological chemistry [J Biol Chem] 2021 Jul; Vol. 297 (1), pp. 100811. Date of Electronic Publication: 2021 May 21. - Publication Year :
- 2021
-
Abstract
- A20 is a potent anti-inflammatory protein that mediates both inflammation and ubiquitination in mammals, but the related mechanisms are not clear. In this study, we performed mass spectrometry (MS) screening, gene ontology (GO) analysis, and coimmunoprecipitation (co-IP) in a lipopolysaccharide (LPS)-induced inflammatory cell model to identify novel A20-interacting proteins. We confirmed that the E3 ubiquitin ligase Nrdp1, also known as ring finger protein 41 (RNF41), interacted with A20 in LPS-stimulated cells. Further co-IP analysis demonstrated that when A20 was knocked out, degradation-inducing K48-linked ubiquitination of inflammatory effector MyD88 was decreased, but protein interaction-mediating K63-linked ubiquitination of another inflammatory effector TBK1 was increased. Moreover, western blot experiments showed that A20 inhibition induced an increase in levels of MyD88 and phosphorylation of downstream effector proteins as well as of TBK1 and a downstream effector, while Nrdp1 inhibition induced an increase in MyD88 but a decrease in TBK1 levels. When A20 and Nrdp1 were coinhibited, no further change in MyD88 was observed, but TBK1 levels were significantly decreased compared with those upon A20 inhibition alone. Gain- and loss-of-function analyses revealed that the ZnF4 domain of A20 is required for Nrdp1 polyubiquitination. Upon LPS stimulation, the inhibition of Nrdp1 alone increased the secretion of IL-6 and TNF-α but decreased IFN-β secretion, as observed in other studies, suggesting that Nrdp1 preferentially promotes the production of IFN-β. Taken together, these results demonstrated that A20/Nrdp1 interaction is important for A20 anti-inflammation, thus revealing a novel mechanism for the anti-inflammatory effects of A20.<br />Competing Interests: Conflict of interest The authors declare that they have no conflicts of interest with the contents of this article.<br /> (Copyright © 2021 The Authors. Published by Elsevier Inc. All rights reserved.)
- Subjects :
- Animals
Enzyme Activation
Inflammation pathology
Interferons metabolism
Macrophages metabolism
Mice
Models, Biological
Protein Binding
Protein Domains
Proteolysis
RAW 264.7 Cells
Signal Transduction
Tumor Necrosis Factor alpha-Induced Protein 3 chemistry
Inflammation metabolism
Lysine metabolism
Myeloid Differentiation Factor 88 metabolism
Polyubiquitin metabolism
Protein Serine-Threonine Kinases metabolism
Tumor Necrosis Factor alpha-Induced Protein 3 metabolism
Ubiquitin-Protein Ligases metabolism
Ubiquitination
Subjects
Details
- Language :
- English
- ISSN :
- 1083-351X
- Volume :
- 297
- Issue :
- 1
- Database :
- MEDLINE
- Journal :
- The Journal of biological chemistry
- Publication Type :
- Academic Journal
- Accession number :
- 34023381
- Full Text :
- https://doi.org/10.1016/j.jbc.2021.100811