title: Ubiquitin-specific Protease 20 Regulates the Reciprocal Functions of β-Arrestin2 in Toll-like Receptor 4-promoted Nuclear Factor κB (NFκB) Activation.
abstract: Toll-like receptor 4 (TLR4) promotes vascular inflammatory disorders such as neointimal hyperplasia and atherosclerosis. TLR4 triggers NFκB signaling through the ubiquitin ligase TRAF6 (tumor necrosis factor receptor-associated factor 6). TRAF6 activity can be impeded by deubiquitinating enzymes like ubiquitin-specific protease 20 (USP20), which can reverse TRAF6 autoubiquitination, and by association with the multifunctional adaptor protein β-arrestin2. Although β-arrestin2 effects on TRAF6 suggest an anti-inflammatory role, physiologic β-arrestin2 promotes inflammation in atherosclerosis and neointimal hyperplasia. We hypothesized that anti- and proinflammatory dimensions of β-arrestin2 activity could be dictated by β-arrestin2's ubiquitination status, which has been linked with its ability to scaffold and localize activated ERK1/2 to signalosomes. With purified proteins and in intact cells, our protein interaction studies showed that TRAF6/USP20 association and subsequent USP20-mediated TRAF6 deubiquitination were β-arrestin2-dependent. Generation of transgenic mice with smooth muscle cell-specific expression of either USP20 or its catalytically inactive mutant revealed anti-inflammatory effects of USP20in vivoandin vitro Carotid endothelial denudation showed that antagonizing smooth muscle cell USP20 activity increased NFκB activation and neointimal hyperplasia. We found that β-arrestin2 ubiquitination was promoted by TLR4 and reversed by USP20. The association of USP20 with β-arrestin2 was augmented when β-arrestin2 ubiquitination was prevented and reduced when β-arrestin2 ubiquitination was rendered constitutive. Constitutive β-arrestin2 ubiquitination also augmented NFκB activation. We infer that pro- and anti-inflammatory activities of β-arrestin2 are determined by β-arrestin2 ubiquitination and that changes in USP20 expression and/or activity can therefore regulate inflammatory responses, at least in part, by defining the ubiquitination status of β-arrestin2.
authors.affiliation: From the Departments of Medicine (Cardiology) and.
authors.forename: Pierre-Yves
authors.initials: PY
authors.lastname: Jean-Charles
authors.affiliation: From the Departments of Medicine (Cardiology) and.
authors.forename: Lisheng
authors.initials: L
authors.lastname: Zhang
authors.affiliation: From the Departments of Medicine (Cardiology) and.
authors.forename: Jiao-Hui
authors.initials: JH
authors.lastname: Wu
authors.affiliation: From the Departments of Medicine (Cardiology) and.
authors.forename: Sang-Oh
authors.initials: SO
authors.lastname: Han
authors.affiliation: From the Departments of Medicine (Cardiology) and.
authors.forename: Leigh
authors.initials: L
authors.lastname: Brian
authors.affiliation: From the Departments of Medicine (Cardiology) and Cell Biology, Duke University Medical Center, Durham, North Carolina 27710 neil.freedman@dm.duke.edu.
authors.forename: Neil J
authors.initials: NJ
authors.lastname: Freedman
authors.affiliation: From the Departments of Medicine (Cardiology) and Cell Biology, Duke University Medical Center, Durham, North Carolina 27710 skshenoy@dm.duke.edu.
authors.forename: Sudha K
authors.initials: SK
authors.lastname: Shenoy
mesh_terms: D000818:Animals; D019390:Arrestins / Q000235:genetics / Q000378:metabolism*; D002460:Cell Line; D010450:Endopeptidases / Q000235:genetics / Q000378:metabolism*; D051379:Mice; D018345:Mice, Knockout; D016328:NF-kappa B / Q000235:genetics / Q000378:metabolism*; D048029:TNF Receptor-Associated Factor 6 / Q000235:genetics / Q000378:metabolism; D051197:Toll-Like Receptor 4 / Q000235:genetics / Q000378:metabolism*; D043222:Ubiquitin Thiolesterase; D054875:Ubiquitination / Q000502:physiology*; D000071557:beta-Arrestins
chemical_list: D019390:Arrestins; D016328:NF-kappa B; D048029:TNF Receptor-Associated Factor 6; C493487:Tlr4 protein, mouse; D051197:Toll-Like Receptor 4; D000071557:beta-Arrestins; D010450:Endopeptidases; D043222:Ubiquitin Thiolesterase; C486762:Usp20 protein, mouse
keywords: NFκB; TNF receptor-associated factor (TRAF); Toll-like receptor 4 (TLR4); USP20; deubiquitylation (deubiquitination); inflammation; neointimal hyperplasia; scaffold; ubiquitin; β-arrestin
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