TY - JOUR
T1 - Site-specific ubiquitination of the E3 ligase HOIP regulates apoptosis and immune signaling
AU - Fennell, Lilian M.
AU - Gomez Diaz, Carlos
AU - Deszcz, Luiza
AU - Kavirayani, Anoop
AU - Hoffmann, David
AU - Yanagitani, Kota
AU - Schleiffer, Alexander
AU - Mechtler, Karl
AU - Hagelkruys, Astrid
AU - Penninger, Josef
AU - Ikeda, Fumiyo
N1 - Funding Information:
We thank Merle Hantsche‐Grininger (EMBL, Heidelberg) and Michaela Morlock (2016 VBC summer student, currently AstraZeneca, Göteborg, Sweden) for their initial contribution to the project setups, Katrin Rittinger (Crick Institute, London) and Paul Elliott (University of Oxford, Oxford) for discussions on the structural aspect of HOIP, and all the Ikeda Laboratory members for constructive discussions and suggestions on the project, as well as the team work, especially Katrin Schodl (IMBA), Olga Olszanska (IMBA) and Akiko Okumura (Kyushu University) also for technical support. We also thank Anna Szydłowska and Kikue Tachibana (IMBA, Vienna) for the technical advice on CRISPR‐Cas9‐based knock‐in mouse generation, Adrian Ting (Icahn School of Medicine at Mount Sinai, New York) for advice on ear‐derived fibroblast isolation, Richard Imre (Protein Chemistry, Vienna) for the Mass spectrometry data analysis, Margit Jaschke (Thermo Scientific, Vienna) for supports with the Cartoplex assays, the IMP‐IMBA core facilities of transgenic service, comparative medicine, molecular biology service, biooptics (especially Pawel Pasierbek), and VBCF facilities, ProTech, and HistoPathology for their technical assistance. Work in the Mechtler laboratory has been supported by EPIC‐XS, project number 823839, funded by the Horizon 2020 Program of the European Union and the by the ERA‐CAPS I 3686 project of the Austrian Science Fund. JMP is supported by the von Zastrow Foundation and a Canada 150 chair. Research in Ikeda Laboratory is supported by JSPS KAKENHI Grant Number 18K19959, ERC Consolidator Grant (LUbi, 614711), FWF stand‐alone grant (P 2550 8) and Austrian Academy of Sciences. We also thank Angela Andersen from the Life Science Editors for editing the manuscript.
Funding Information:
We thank Merle Hantsche-Grininger (EMBL, Heidelberg) and Michaela Morlock (2016 VBC summer student, currently AstraZeneca, G?teborg, Sweden) for their initial contribution to the project setups, Katrin Rittinger (Crick Institute, London) and Paul Elliott (University of Oxford, Oxford) for discussions on the structural aspect of HOIP, and all the Ikeda Laboratory members for constructive discussions and suggestions on the project, as well as the team work, especially Katrin Schodl (IMBA), Olga Olszanska (IMBA) and Akiko Okumura (Kyushu University) also for technical support. We also thank Anna Szyd?owska and Kikue Tachibana (IMBA, Vienna) for the technical advice on CRISPR-Cas9-based knock-in mouse generation, Adrian Ting (Icahn School of Medicine at Mount Sinai, New York) for advice on ear-derived fibroblast isolation, Richard Imre (Protein Chemistry, Vienna) for the Mass spectrometry data analysis, Margit Jaschke (Thermo Scientific, Vienna) for supports with the Cartoplex assays, the IMP-IMBA core facilities of transgenic service, comparative medicine, molecular biology service, biooptics (especially Pawel Pasierbek), and VBCF facilities, ProTech, and HistoPathology for their technical assistance. Work in the Mechtler laboratory has been supported by EPIC-XS, project number 823839, funded by the Horizon 2020 Program of the European Union and the by the ERA-CAPS I 3686 project of the Austrian Science Fund. JMP is supported by the von Zastrow Foundation and a Canada 150 chair. Research in Ikeda Laboratory is supported by JSPS KAKENHI Grant Number 18K19959, ERC Consolidator Grant (LUbi, 614711), FWF stand-alone grant (P 2550 8) and Austrian Academy of Sciences. We also thank Angela Andersen from the Life Science Editors for editing the manuscript.
Publisher Copyright:
© 2020 The Authors
PY - 2020/12/15
Y1 - 2020/12/15
N2 - HOIP, the catalytic component of the linear ubiquitin chain assembly complex (LUBAC), is a critical regulator of inflammation. However, how HOIP itself is regulated to control inflammatory responses is unclear. Here, we discover that site-specific ubiquitination of K784 within human HOIP promotes tumor necrosis factor (TNF)-induced inflammatory signaling. A HOIP K784R mutant is catalytically active but shows reduced induction of an NF-κB reporter relative to wild-type HOIP. HOIP K784 is evolutionarily conserved, equivalent to HOIP K778 in mice. We generated HoipK778R/K778R knock-in mice, which show no overt developmental phenotypes; however, in response to TNF, HoipK778R/K778R mouse embryonic fibroblasts display mildly suppressed NF-κB activation and increased apoptotic markers. On the other hand, HOIP K778R enhances the TNF-induced formation of TNFR complex II and an interaction between TNFR complex II and LUBAC. Loss of the LUBAC component SHARPIN leads to embryonic lethality in HoipK778R/K778R mice, which is rescued by knockout of TNFR1. We propose that site-specific ubiquitination of HOIP regulates a LUBAC-dependent switch between survival and apoptosis in TNF signaling.
AB - HOIP, the catalytic component of the linear ubiquitin chain assembly complex (LUBAC), is a critical regulator of inflammation. However, how HOIP itself is regulated to control inflammatory responses is unclear. Here, we discover that site-specific ubiquitination of K784 within human HOIP promotes tumor necrosis factor (TNF)-induced inflammatory signaling. A HOIP K784R mutant is catalytically active but shows reduced induction of an NF-κB reporter relative to wild-type HOIP. HOIP K784 is evolutionarily conserved, equivalent to HOIP K778 in mice. We generated HoipK778R/K778R knock-in mice, which show no overt developmental phenotypes; however, in response to TNF, HoipK778R/K778R mouse embryonic fibroblasts display mildly suppressed NF-κB activation and increased apoptotic markers. On the other hand, HOIP K778R enhances the TNF-induced formation of TNFR complex II and an interaction between TNFR complex II and LUBAC. Loss of the LUBAC component SHARPIN leads to embryonic lethality in HoipK778R/K778R mice, which is rescued by knockout of TNFR1. We propose that site-specific ubiquitination of HOIP regulates a LUBAC-dependent switch between survival and apoptosis in TNF signaling.
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U2 - 10.15252/embj.2019103303
DO - 10.15252/embj.2019103303
M3 - Article
C2 - 33215740
AN - SCOPUS:85096747636
VL - 39
JO - EMBO Journal
JF - EMBO Journal
SN - 0261-4189
IS - 24
M1 - e103303
ER -