Journal article

MLKL trafficking and accumulation at the plasma membrane control the kinetics and threshold for necroptosis

Andre L Samson, Ying Zhang, Niall D Geoghegan, Xavier J Gavin, Katherine A Davies, Michael J Mlodzianoski, Lachlan W Whitehead, Daniel Frank, Sarah E Garnish, Cheree Fitzgibbon, Anne Hempel, Samuel N Young, Annette V Jacobsen, Wayne Cawthorne, Emma J Petrie, Maree C Faux, Kristy Shield-Artin, Najoua Lalaoui, Joanne M Hildebrand, John Silke Show all



Mixed lineage kinase domain-like (MLKL) is the terminal protein in the pro-inflammatory necroptotic cell death program. RIPK3-mediated phosphorylation is thought to initiate MLKL oligomerization, membrane translocation and membrane disruption, although the precise choreography of events is incompletely understood. Here, we use single-cell imaging approaches to map the chronology of endogenous human MLKL activation during necroptosis. During the effector phase of necroptosis, we observe that phosphorylated MLKL assembles into higher order species on presumed cytoplasmic necrosomes. Subsequently, MLKL co-traffics with tight junction proteins to the cell periphery via Golgi-microtubule-actin-de..

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Awarded by Australian National Health and Medical Research Council

Awarded by Victorian Cancer Agency

Funding Acknowledgements

We thank the Walter and Eliza Hall Institute Monoclonal Antibody Facility for their assistance generating the anti-human MLKL antibodies. We are grateful to the Australian National Health and Medical Research Council for fellowship (J. M.H., 1142669; G.L., 1117089; E.D.H., 1159488; J.M.M., 1105754 and 1172929), grant (1124735, 1124737, and 1105023), and infrastructure (IRIISS 9000587) support and the Victorian Cancer Agency for fellowship support (N.L., 17030), with additional support from the CASS Foundation (A.L.S.), the Australian Cancer Research Foundation and the Victorian Government Operational Infrastructure Support scheme. We acknowledge the support for A.V.J., S.E.G. and K.A.D. from an Australian Research Training Program Scholarship, the support for D.F. from a Melbourne Research Scholarship, and additional support for K.A.D. by an AINSE postgraduate research award. The Lattice light sheet referenced in this research was used under license from Howard Hughes Medical Institute, Janelia Research Campus. We thank Prof. Matt Sweet (University of Queensland) for the MLKL2 isoform cDNA. We thank Ueli Nachbur and Antony W. Burgess for support and helpful comments about the manuscript. mTagRFP-Membrane-1 was a gift from Michael Davidson (The National High Magnetic Field Laboratory; Addgene plasmid #57992;; RRID:Addgene_57992).