Phosphorylation of USP20 on Ser334 by IRAK1 promotes IL-1β-evoked signaling in vascular smooth muscle cells and vascular inflammation
Copyright © 2023 The Authors. Published by Elsevier Inc. All rights reserved..
Reversible lysine-63 (K63) polyubiquitination regulates proinflammatory signaling in vascular smooth muscle cells (SMCs) and plays an integral role in atherosclerosis. Ubiquitin-specific peptidase 20 (USP20) reduces NFκB activation triggered by proinflammatory stimuli, and USP20 activity attenuates atherosclerosis in mice. The association of USP20 with its substrates triggers deubiquitinase activity; this association is regulated by phosphorylation of USP20 on Ser334 (mouse) or Ser333 (human). USP20 Ser333 phosphorylation was greater in SMCs of atherosclerotic segments of human arteries as compared with nonatherosclerotic segments. To determine whether USP20 Ser334 phosphorylation regulates proinflammatory signaling, we created USP20-S334A mice using CRISPR/Cas9-mediated gene editing. USP20-S334A mice developed ∼50% less neointimal hyperplasia than congenic WT mice after carotid endothelial denudation. WT carotid SMCs showed substantial phosphorylation of USP20 Ser334, and WT carotids demonstrated greater NFκB activation, VCAM-1 expression, and SMC proliferation than USP20-S334A carotids. Concordantly, USP20-S334A primary SMCs in vitro proliferated and migrated less than WT SMCs in response to IL-1β. An active site ubiquitin probe bound to USP20-S334A and USP20-WT equivalently, but USP20-S334A associated more avidly with TRAF6 than USP20-WT. IL-1β induced less K63-linked polyubiquitination of TRAF6 and less downstream NFκB activity in USP20-S334A than in WT SMCs. Using in vitro phosphorylation with purified IRAK1 and siRNA-mediated gene silencing of IRAK1 in SMCs, we identified IRAK1 as a novel kinase for IL-1β-induced USP20 Ser334 phosphorylation. Our findings reveal novel mechanisms regulating IL-1β-induced proinflammatory signaling: by phosphorylating USP20 Ser334, IRAK1 diminishes the association of USP20 with TRAF6 and thus augments NFκB activation, SMC inflammation, and neointimal hyperplasia.
| Medienart: |
E-Artikel |
|---|
| Erscheinungsjahr: |
2023 |
|---|---|
| Erschienen: |
2023 |
| Enthalten in: |
Zur Gesamtaufnahme - volume:299 |
|---|---|
| Enthalten in: |
The Journal of biological chemistry - 299(2023), 7 vom: 03. Juli, Seite 104911 |
| Sprache: |
Englisch |
|---|
| Beteiligte Personen: |
Zhang, Lisheng [VerfasserIn] |
|---|
| Links: |
|---|
| Anmerkungen: |
Date Completed 07.08.2023 Date Revised 13.04.2024 published: Print-Electronic Citation Status MEDLINE |
|---|
| doi: |
10.1016/j.jbc.2023.104911 |
|---|
| funding: |
|
|---|---|
| Förderinstitution / Projekttitel: |
|
| PPN (Katalog-ID): |
NLM358131979 |
|---|
| LEADER | 01000caa a22002652 4500 | ||
|---|---|---|---|
| 001 | NLM358131979 | ||
| 003 | DE-627 | ||
| 005 | 20240413232316.0 | ||
| 007 | cr uuu---uuuuu | ||
| 008 | 231226s2023 xx |||||o 00| ||eng c | ||
| 024 | 7 | |a 10.1016/j.jbc.2023.104911 |2 doi | |
| 028 | 5 | 2 | |a pubmed24n1374.xml |
| 035 | |a (DE-627)NLM358131979 | ||
| 035 | |a (NLM)37311534 | ||
| 035 | |a (PII)S0021-9258(23)01939-7 | ||
| 040 | |a DE-627 |b ger |c DE-627 |e rakwb | ||
| 041 | |a eng | ||
| 100 | 1 | |a Zhang, Lisheng |e verfasserin |4 aut | |
| 245 | 1 | 0 | |a Phosphorylation of USP20 on Ser334 by IRAK1 promotes IL-1β-evoked signaling in vascular smooth muscle cells and vascular inflammation |
| 264 | 1 | |c 2023 | |
| 336 | |a Text |b txt |2 rdacontent | ||
| 337 | |a ƒaComputermedien |b c |2 rdamedia | ||
| 338 | |a ƒa Online-Ressource |b cr |2 rdacarrier | ||
| 500 | |a Date Completed 07.08.2023 | ||
| 500 | |a Date Revised 13.04.2024 | ||
| 500 | |a published: Print-Electronic | ||
| 500 | |a Citation Status MEDLINE | ||
| 520 | |a Copyright © 2023 The Authors. Published by Elsevier Inc. All rights reserved. | ||
| 520 | |a Reversible lysine-63 (K63) polyubiquitination regulates proinflammatory signaling in vascular smooth muscle cells (SMCs) and plays an integral role in atherosclerosis. Ubiquitin-specific peptidase 20 (USP20) reduces NFκB activation triggered by proinflammatory stimuli, and USP20 activity attenuates atherosclerosis in mice. The association of USP20 with its substrates triggers deubiquitinase activity; this association is regulated by phosphorylation of USP20 on Ser334 (mouse) or Ser333 (human). USP20 Ser333 phosphorylation was greater in SMCs of atherosclerotic segments of human arteries as compared with nonatherosclerotic segments. To determine whether USP20 Ser334 phosphorylation regulates proinflammatory signaling, we created USP20-S334A mice using CRISPR/Cas9-mediated gene editing. USP20-S334A mice developed ∼50% less neointimal hyperplasia than congenic WT mice after carotid endothelial denudation. WT carotid SMCs showed substantial phosphorylation of USP20 Ser334, and WT carotids demonstrated greater NFκB activation, VCAM-1 expression, and SMC proliferation than USP20-S334A carotids. Concordantly, USP20-S334A primary SMCs in vitro proliferated and migrated less than WT SMCs in response to IL-1β. An active site ubiquitin probe bound to USP20-S334A and USP20-WT equivalently, but USP20-S334A associated more avidly with TRAF6 than USP20-WT. IL-1β induced less K63-linked polyubiquitination of TRAF6 and less downstream NFκB activity in USP20-S334A than in WT SMCs. Using in vitro phosphorylation with purified IRAK1 and siRNA-mediated gene silencing of IRAK1 in SMCs, we identified IRAK1 as a novel kinase for IL-1β-induced USP20 Ser334 phosphorylation. Our findings reveal novel mechanisms regulating IL-1β-induced proinflammatory signaling: by phosphorylating USP20 Ser334, IRAK1 diminishes the association of USP20 with TRAF6 and thus augments NFκB activation, SMC inflammation, and neointimal hyperplasia | ||
| 650 | 4 | |a Journal Article | |
| 650 | 4 | |a Research Support, N.I.H., Extramural | |
| 650 | 4 | |a Research Support, Non-U.S. Gov't | |
| 650 | 4 | |a IRAK1 | |
| 650 | 4 | |a NFκB | |
| 650 | 4 | |a TRAF6 | |
| 650 | 4 | |a de-ubiquitination | |
| 650 | 4 | |a interleukin-1β | |
| 650 | 4 | |a vascular injury | |
| 650 | 7 | |a IL1B protein, mouse |2 NLM | |
| 650 | 7 | |a Interleukin-1 Receptor-Associated Kinases |2 NLM | |
| 650 | 7 | |a EC 2.7.11.1 |2 NLM | |
| 650 | 7 | |a Irak1 protein, mouse |2 NLM | |
| 650 | 7 | |a EC 2.7.11.1 |2 NLM | |
| 650 | 7 | |a Phosphoserine |2 NLM | |
| 650 | 7 | |a 17885-08-4 |2 NLM | |
| 650 | 7 | |a TNF Receptor-Associated Factor 6 |2 NLM | |
| 650 | 7 | |a Ubiquitin Thiolesterase |2 NLM | |
| 650 | 7 | |a EC 3.4.19.12 |2 NLM | |
| 650 | 7 | |a USP20 protein, human |2 NLM | |
| 650 | 7 | |a Usp20 protein, mouse |2 NLM | |
| 650 | 7 | |a EC 3.4.19.12 |2 NLM | |
| 650 | 7 | |a NF-kappa B |2 NLM | |
| 650 | 7 | |a Interleukin-1beta |2 NLM | |
| 700 | 1 | |a Wu, Jiao-Hui |e verfasserin |4 aut | |
| 700 | 1 | |a Jean-Charles, Pierre-Yves |e verfasserin |4 aut | |
| 700 | 1 | |a Murali, Pavitra |e verfasserin |4 aut | |
| 700 | 1 | |a Zhang, Wenli |e verfasserin |4 aut | |
| 700 | 1 | |a Jazic, Aeva |e verfasserin |4 aut | |
| 700 | 1 | |a Kaur, Suneet |e verfasserin |4 aut | |
| 700 | 1 | |a Nepliouev, Igor |e verfasserin |4 aut | |
| 700 | 1 | |a Stiber, Jonathan A |e verfasserin |4 aut | |
| 700 | 1 | |a Snow, Kamie |e verfasserin |4 aut | |
| 700 | 1 | |a Freedman, Neil J |e verfasserin |4 aut | |
| 700 | 1 | |a Shenoy, Sudha K |e verfasserin |4 aut | |
| 773 | 0 | 8 | |i Enthalten in |t The Journal of biological chemistry |d 1945 |g 299(2023), 7 vom: 03. Juli, Seite 104911 |w (DE-627)NLM000004995 |x 1083-351X |7 nnns |
| 773 | 1 | 8 | |g volume:299 |g year:2023 |g number:7 |g day:03 |g month:07 |g pages:104911 |
| 856 | 4 | 0 | |u http://dx.doi.org/10.1016/j.jbc.2023.104911 |3 Volltext |
| 912 | |a GBV_USEFLAG_A | ||
| 912 | |a GBV_NLM | ||
| 951 | |a AR | ||
| 952 | |d 299 |j 2023 |e 7 |b 03 |c 07 |h 104911 | ||