Ubiquitylation of MLKL at lysine 219 positively regulates necroptosis-induced tissue injury and pathogen clearance

• Published in: Nature communications Vol. 12; no. 1; pp. 3364 - 18
• Main Authors: Garcia, Laura Ramos, Tenev, Tencho, Newman, Richard, Haich, Rachel O., Liccardi, Gianmaria, John, Sidonie Wicky, Annibaldi, Alessandro, Yu, Lu, Pardo, Mercedes, Young, Samuel N., Fitzgibbon, Cheree, Fernando, Winnie, Guppy, Naomi, Kim, Hyojin, Liang, Lung-Yu, Lucet, Isabelle S., Kueh, Andrew, Roxanis, Ioannis, Gazinska, Patrycja, Sims, Martin, Smyth, Tomoko, Ward, George, Bertin, John, Beal, Allison M., Geddes, Brad, Choudhary, Jyoti S., Murphy, James M., Aurelia Ball, K., Upton, Jason W., Meier, Pascal
• Format: Journal Article
• Language: English
• Published: London Nature Publishing Group UK 07.06.2021; Nature Publishing Group; Nature Portfolio
• Subjects: 13/106; 14/34; 38/70; 631/337/458/582; 631/80/458/582; 631/80/82/2344; 64/60; 82/58; 96/1; Animals; Apoptosis; Assembly; Cell death; Cell Line; Cells, Cultured; Clearances; Cytotoxicity; HEK293 Cells; Herpesviridae Infections - genetics; Herpesviridae Infections - metabolism; Herpesviridae Infections - virology; HT29 Cells; Humanities and Social Sciences; Humans; Inflammation; Lysine; Lysine - genetics; Lysine - metabolism; Membranes; Mice; Mice, Inbred C57BL; Mice, Knockout; Mortality; multidisciplinary; Muromegalovirus - physiology; Mutation; Necroptosis; Necroptosis - genetics; Necrosis; NIH 3T3 Cells; Oligomerization; Pathogenesis; Pathogens; Phosphorylation; Protein Kinases - genetics; Protein Kinases - metabolism; Rupture; Science; Science (multidisciplinary); Signalling systems; Skin - metabolism; Skin - pathology; Toxicity; Translocation; Ubiquitin; Ubiquitination
• ISSN: 2041-1723; 2041-1723
• DOI: 10.1038/s41467-021-23474-5

Necroptosis is a lytic, inflammatory form of cell death that not only contributes to pathogen clearance but can also lead to disease pathogenesis. Necroptosis is triggered by RIPK3-mediated phosphorylation of MLKL, which is thought to initiate MLKL oligomerisation, membrane translocation and membrane rupture, although the precise mechanism is incompletely understood. Here, we show that K63-linked ubiquitin chains are attached to MLKL during necroptosis and that ubiquitylation of MLKL at K219 significantly contributes to the cytotoxic potential of phosphorylated MLKL. The K219R MLKL mutation protects animals from necroptosis-induced skin damage and renders cells resistant to pathogen-induced necroptosis. Mechanistically, we show that ubiquitylation of MLKL at K219 is required for higher-order assembly of MLKL at membranes, facilitating its rupture and necroptosis. We demonstrate that K219 ubiquitylation licenses MLKL activity to induce lytic cell death, suggesting that necroptotic clearance of pathogens as well as MLKL-dependent pathologies are influenced by the ubiquitin-signalling system. Necroptosis is a form of cell death characterized by membrane rupture via MLKL oligomerization, although mechanistic details remain unclear. Here, the authors show that MLKL ubiquitylation of K219 facilitates high-order membrane assembly and subsequent rupture, promoting cytotoxicity.