Activation of the pseudokinase MLKL unleashes the four-helix bundle domain to induce membrane localization and necroptotic cell death

JM Hildebrand; MC Tanzer; IS Lucet; SN Young; SK Spall; P Sharma; C Pierotti; JM Garnier; RCJ Dobson; AI Webb; A Tripaydonis; JJ Babon; MD Mulcair; MJ Scanlon; WS Alexander; AF Wilks; PE Czabotar; G Lessene; JM Murphy; J Silke

Journal article

Activation of the pseudokinase MLKL unleashes the four-helix bundle domain to induce membrane localization and necroptotic cell death

JM Hildebrand, MC Tanzer, IS Lucet, SN Young, SK Spall, P Sharma, C Pierotti, JM Garnier, RCJ Dobson, AI Webb, A Tripaydonis, JJ Babon, MD Mulcair, MJ Scanlon, WS Alexander, AF Wilks, PE Czabotar, G Lessene, JM Murphy, J Silke

Proceedings of the National Academy of Sciences of the United States of America | Published : 2014

DOI: 10.1073/pnas.1408987111

Abstract

Necroptosis is considered to be complementary to the classical caspase-dependent programmed cell death pathway, apoptosis. The pseudokinase Mixed Lineage Kinase Domain-Like (MLKL) is an essential effector protein in the necroptotic cell death pathway downstream of the protein kinase Receptor Interacting Protein Kinase-3 (RIPK3). How MLKL causes cell death is unclear, however RIPK3-mediated phosphorylation of the activation loop in MLKL trips a molecular switch to induce necroptotic cell death. Here, we show that the MLKL pseudokinase domain acts as a latch to restrain the N-terminal four-helix bundle (4HB) domain and that unleashing this domain results in formation of a high-molecular-weight..

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University of Melbourne Researchers

Joanne Hildebrand Author

Isabelle Lucet Author

Renwick Dobson Author

Andrew Webb Author

Grants

Funding Acknowledgements

We thank staff in the Walter and Eliza Hall Institute Bioservices Facility; Drs. Robert Ninnis, Grant Dewson, and James Vince for advice; Vishva Dixit for Ripk3<SUP>-/-</SUP> mice; and Toru Okamoto for developing inducible lentiviral vectors. This work was supported by National Health and Medical Research Council (NHMRC) Grants 1057905, 1067289, 1046984, 1025594, and 461221 and fellowships (to J.M.H., W.S.A., and J.S.); a Victorian International Research Scholarship (to M.C.T.); a C. R. Roper Fellowship (to R.C.J.D.); and Australian Research Council fellowships (to J.J.B., P.E.C., and J.M.M.). Additional support was obtained from the Australian Cancer Research Fund, Victorian State Government Operational Infrastructure Support, and NHMRC Independent Research Institute Infrastructure Support Grant 361646.