Journal article

A missense mutation in the MLKL brace region promotes lethal neonatal inflammation and hematopoietic dysfunction

Joanne M Hildebrand, Maria Kauppi, Ian J Majewski, Zikou Liu, Allison J Cox, Sanae Miyake, Emma J Petrie, Michael A Silk, Zhixiu Li, Maria C Tanzer, Gabriela Brumatti, Samuel N Young, Cathrine Hall, Sarah E Garnish, Jason Corbin, Michael D Stutz, Ladina Di Rago, Pradnya Gangatirkar, Emma C Josefsson, Kristin Rigbye Show all

Nature Communications | NATURE PUBLISHING GROUP | Published : 2020

Abstract

MLKL is the essential effector of necroptosis, a form of programmed lytic cell death. We have isolated a mouse strain with a single missense mutation, MlklD139V, that alters the two-helix 'brace' that connects the killer four-helix bundle and regulatory pseudokinase domains. This confers constitutive, RIPK3 independent killing activity to MLKL. Homozygous mutant mice develop lethal postnatal inflammation of the salivary glands and mediastinum. The normal embryonic development of MlklD139V homozygotes until birth, and the absence of any overt phenotype in heterozygotes provides important in vivo precedent for the capacity of cells to clear activated MLKL. These observations offer an important..

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Grants

Awarded by Australian National Health and Medical Research Council (NHMRC)


Awarded by NHMRC


Awarded by NIH


Awarded by National Institute of Arthritis and Musculoskeletal and Skin Diseases (NIAMS) at the National Institutes of Health


Awarded by Japan Society for the Promotion of Science, and Scientific Research on Innovative areas


Awarded by Japan Agency for Medical Research and Development (AMED) through AMED-CREST


Awarded by Jack Brockhoff Foundation


Awarded by NHMRC Project and Targeted Research grants


Awarded by Leukemia Foundation Australia priority grant


Awarded by Victoria Cancer Agency (VCA) mid-career fellowship


Awarded by Arthritis Research UK


Awarded by Wellcome Trust


Awarded by Oxford Comprehensive Biomedical Research Centre ankylosing spondylitis chronic disease cohort


Funding Acknowledgements

We thank all the following people for their technical assistance; Jiami Han, Cynthia Liu, Jasmine McManus, Janelle Lochland and Emma Tovey (WEHI), Aira Nuguid and Tina Cardamone (APN histopathology - The University of Melbourne). Thomas Boudier (WEHI Centre for Dynamic Imaging). The WEHI Histology Service, WEHI Antibody Facility and WEHI Bioservices. Y. Uchiyama and S. Kakuta who advised the interpretation of the results of TEM and Annette Jacobsen (WEHI) for important insight and discussion. The generation of Mlkl<SUP>D139V</SUP> mice by CRISPR/Cas9 was performed by Andrew Kueh and Marco Herold (WEHI MAGEC laboratory) supported by the Australian Phenomics Network (APN) and the Australian Government through the National Collaborative Research Infrastructure Strategy (NCRIS) program. This work is supported by; Project grant (1105023) and Fellowships (0541951 and 1142669) from the Australian National Health and Medical Research Council (NHMRC) to J.M.H.; Fellowship (1107149) from the NHMRC to J.S.; Program grant (1113577) and Fellowship (1058344) from the NHMRC (W.S.A.); Project grant (1124735) and Fellowships (1105754, 1172929) from the NHMRC (J.M.M.); NIH training grants T32GM008629 and T32GM082729-01 (A.J.C.); R01AR059703 from the National Institute of Arthritis and Musculoskeletal and Skin Diseases (NIAMS) at the National Institutes of Health (P.J.F. and A.G.B.); the Marjorie K. Lamb Professorship (P.J.F.); Grants-in-Aid from Scientific Research (B) 17H04069 from Japan Society for the Promotion of Science, and Scientific Research on Innovative areas 26110003, the Japan Agency for Medical Research and Development (AMED) through AMED-CREST JP20gm1210002 and Private University Research Branding project from a MEXT (H.N.); Victorian International Research Scholarship (Z.L. and M.C.T.). Australia Postgraduate Award (C.A.D.); S.L.M. acknowledges funding from NHMRC grants (1144282, 1142354, and 1099262), The Sylvia and Charles Viertel Foundation, HHMI-Wellcome International Research Scholarship and Glaxosmithkline; Fellowship from the Lorenzo and Pamela Galli Charitable Trust (E.C.J.), NHMRC grants 1107425 and 1045549 and The Sylvia & Charles Viertel Senior Medical Research Fellowship (M.P.); D.B.A. was supported by the Jack Brockhoff Foundation (JBF 4186, 2016) and NHMRC Fellowship (APP1072476); NHMRC IRIISS 9000587 and Victorian Government Operational Infrastructure Support schemes; NHMRC Project and Targeted Research grants 1006769, 512672, and 512381 (M.F.S.); The Department of Industry, Innovation, Science, Research and Tertiary Education Collaborative Research Network and Diabetes Australia (MAB); I.J.M. was supported by the Victorian Cancer Agency, and by generous support from the Felton Bequest; Cancer Australia and Leukemia Foundation Australia priority grant (PdCCRS 1162023) and a Victoria Cancer Agency (VCA) mid-career fellowship (MCRF 15027) to G.B. We gratefully acknowledge the contribution of genotype data by Dr Yorgi Mavros (University of Sydney), Professor Nick Martin (QIMR), Professor Jim Rosenbaum (Oregon Health and Science University), and Professor Maxime Breban and the Groupe Francaise d'Etude Genetique des Spondylarthrites (GFEGS). We are grateful to Professor BP Wordsworth of the University of Oxford, UK for access to genotype data on ankylosing spondylitis cases collected in studies funded, in part, by Arthritis Research UK (grants 19536 and 18797), by the Wellcome Trust (grant 076113) and by the Oxford Comprehensive Biomedical Research Centre ankylosing spondylitis chronic disease cohort (theme A91202).