TY - JOUR
T1 - A20 protects cells from TNF-induced apoptosis through linear ubiquitin-dependent and -independent mechanisms
AU - Priem, Dario
AU - Devos, Michael
AU - Druwé, Sarah
AU - Martens, Arne
AU - Slowicka, Karolina
AU - Ting, Adrian T.
AU - Pasparakis, Manolis
AU - Declerq, Wim
AU - Vandenabeele, Peter
AU - van Loo, Geert
AU - Bertrand, Mathieu J.M.
N1 - Funding Information:
We are grateful to V. Dixit and K. Newton (Genentech, USA) for the Ripk3−/− mice, J. Silke (WEHI, Australia) for the ciap1/2−/− MEFs, H. Walczak (UCL Cancer Institute, UK) for the Hoip−/− MEFs, Y. Dikic (Goethe University, Germany) for GST-UBAN plasmids and S. Hofmans and K. Augustyns (University of Antwerp, Belgium) for the synthesis of Nec-1s. We also thank the VIB Screening core facility for excellent assistance. Research in the groups of M.J.M.B., G.v.L., W.D., and P.V. is financially supported by the Vlaams Instituut voor Biotechnologie (VIB), by Ghent University (GROUP-ID, GOA-01G01914), by grants from the Fonds voor Wetenschappelijk Onderzoek Vlaanderen (FWO) (G013715N, G0C3114N, G044518N, EOS MODEL-IDI 30826052, G017212N, G078713N, G0E0416N, G0C7618N, and G0B7118N), from the Belgian science policy office (BELSPO) (IAP 7/32), from the Flemish Government (Methusalem BOF09/ 01M00709 and BOF16/MET_V/007), from the “Foundation against Cancer” (2012-188 and FAF-F/2016/865, FAF-F/2016/868), from the “Geneeskundige Stichting Koningin Elisabeth” (GSKE), from the CBC Banque Prize and from the Charcot Foundation. D.P. was paid by a strategic basic research PhD fellowship from the FWO.
Publisher Copyright:
© 2019, The Author(s).
PY - 2019/10/1
Y1 - 2019/10/1
N2 - The cytokine TNF promotes inflammation either directly by activating the MAPK and NF-κB signaling pathways, or indirectly by triggering cell death. A20 is a potent anti-inflammatory molecule, and mutations in the gene encoding A20 are associated with a wide panel of inflammatory pathologies, both in human and in the mouse. Binding of TNF to TNFR1 triggers the NF-κB-dependent expression of A20 as part of a negative feedback mechanism preventing sustained NF-κB activation. Apart from acting as an NF-κB inhibitor, A20 is also well-known for its ability to counteract the cytotoxic potential of TNF. However, the mechanism by which A20 mediates this function and the exact cell death modality that it represses have remained incompletely understood. In the present study, we provide in vitro and in vivo evidences that deletion of A20 induces RIPK1 kinase-dependent and -independent apoptosis upon single TNF stimulation. We show that constitutively expressed A20 is recruited to TNFR1 signaling complex (Complex I) via its seventh zinc finger (ZF7) domain, in a cIAP1/2-dependent manner, within minutes after TNF sensing. We demonstrate that Complex I-recruited A20 protects cells from apoptosis by stabilizing the linear (M1) ubiquitin network associated to Complex I, a process independent of its E3 ubiquitin ligase and deubiquitylase (DUB) activities and which is counteracted by the DUB CYLD, both in vitro and in vivo. In absence of linear ubiquitylation, A20 is still recruited to Complex I via its ZF4 and ZF7 domains, but this time protects the cells from death by deploying its DUB activity. Together, our results therefore demonstrate two distinct molecular mechanisms by which constitutively expressed A20 protect cells from TNF-induced apoptosis.
AB - The cytokine TNF promotes inflammation either directly by activating the MAPK and NF-κB signaling pathways, or indirectly by triggering cell death. A20 is a potent anti-inflammatory molecule, and mutations in the gene encoding A20 are associated with a wide panel of inflammatory pathologies, both in human and in the mouse. Binding of TNF to TNFR1 triggers the NF-κB-dependent expression of A20 as part of a negative feedback mechanism preventing sustained NF-κB activation. Apart from acting as an NF-κB inhibitor, A20 is also well-known for its ability to counteract the cytotoxic potential of TNF. However, the mechanism by which A20 mediates this function and the exact cell death modality that it represses have remained incompletely understood. In the present study, we provide in vitro and in vivo evidences that deletion of A20 induces RIPK1 kinase-dependent and -independent apoptosis upon single TNF stimulation. We show that constitutively expressed A20 is recruited to TNFR1 signaling complex (Complex I) via its seventh zinc finger (ZF7) domain, in a cIAP1/2-dependent manner, within minutes after TNF sensing. We demonstrate that Complex I-recruited A20 protects cells from apoptosis by stabilizing the linear (M1) ubiquitin network associated to Complex I, a process independent of its E3 ubiquitin ligase and deubiquitylase (DUB) activities and which is counteracted by the DUB CYLD, both in vitro and in vivo. In absence of linear ubiquitylation, A20 is still recruited to Complex I via its ZF4 and ZF7 domains, but this time protects the cells from death by deploying its DUB activity. Together, our results therefore demonstrate two distinct molecular mechanisms by which constitutively expressed A20 protect cells from TNF-induced apoptosis.
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UR - http://www.scopus.com/inward/citedby.url?scp=85072407158&partnerID=8YFLogxK
U2 - 10.1038/s41419-019-1937-y
DO - 10.1038/s41419-019-1937-y
M3 - Article
C2 - 31534131
AN - SCOPUS:85072407158
SN - 2041-4889
VL - 10
JO - Cell Death and Disease
JF - Cell Death and Disease
IS - 10
M1 - 692
ER -