ALS-associated missense and nonsense TBK1 mutations can both cause loss of kinase function

• Type of Material: Article
• Authors: Martina de Majo, Kings College London; Simon D. Topp, Kings College London; Bradley N. Smith, Kings College London; Agnes L. Nishimura, Kings College London; Han-Jou Chen, Kings College London; Athina Soragia Gkazi, Kings College London; Jack Miller, Kings College London; Chun Hao Wong, Kings College London; Caroline Vance, Kings College London; Frank Baas, Leiden University; Anneloor L.M.A ten Asbroek, University of Amsterdam; Kevin P. Kenna, University of Massachusetts; Nicola Ticozzi, IRCCS Istituto Auxologico Italiano; Alberto Garcia Redondo, Instituto de Investigación Hospital 12 de Octubre de Madrid; Jesus Esteban-Perez, Instituto de Investigación Hospital 12 de Octubre de Madrid; Cinzia Tiloca, IRCCS Istituto Auxologico Italiano; Federico Verde, IRCCS Istituto Auxologico Italiano; Stefano Duga, Humanitas University; Karen E. Morrison, University of Southampton; Pamela J. Shaw, University of Sheffield; Janine Kirby, University of Sheffield; Martin R. Turner, University of Oxford; Kevin Talbot, University of Oxford; Orla Hardiman, Trinity College Dublin; Jonathan Glass, Emory University; Jacqueline de Belleroche, Imperial College London; Cinzia Gellera, Fondazione IRCCS Istituto Neurologico 'Carlo Besta'; Antonia Ratti, IRCCS Istituto Auxologico Italiano; Ammar Al-Chalabi, Kings Coll London; Robert H. Brown, University of Massachusetts; Vincenzo Silani, IRCCS Istituto Auxologico Italiano; John E. Landers, University of Massachusetts; Christopher E. Shaw, Kings College London
• Language: English
• Date: 2018-11-01
• Publisher: Elsevier Science Inc.
• Publication Version: Final Published Version
• Copyright Statement: © 2018 The Authors
• License: Creative Commons Attribution 4.0 International
• Final Published Version (URL): http://dx.doi.org/10.1016/j.neurobiolaging.2018.06.015
• Title of Journal or Parent Work: Neurobiology of Aging
• Volume: 71
• Start Page: 266.e1
• End Page: 266.e10
• Grant/Funding Information: Funding was provided to JEL by the National Institutes of Health (NIH)/National Institute of Neurological Disorders and Stroke (NINDS) (R01NS073873) and the American ALS Association.; Samples used in this research were in part obtained from the UK National DNA Bank for MND Research, funded by the MND Association and the Wellcome Trust.; CES and AA-C receive salary support from the National Institute for Health Research (NIHR) Dementia Biomedical Research Unit at South London and Maudsley NHS Foundation Trust and King's College London.; This is an EU Joint Program–Neurodegenerative Disease Research (JPND) project. The project is supported through the following funding organizations under the aegis of JPND–www.jpnd.eu (United Kingdom, Medical Research Council and Economic and Social Research Council) and Horizon 2020 Program (H2020-PHC-2014-two-stage; grant agreement number 633413).; BNS is supported by a Medical Research Fund fellowship.; This work was supported by Heaton-Ellis Trust, Motor Neurone Disease Association, Medical Research Council, the Wellcome Trust, the Noreen Murray Foundation, the American ALS Association, and Project MinE.
• Keywords: FTD; Life Sciences & Biomedicine; Neurosciences & Neurology; OPTINEURIN; OPTN; Geriatrics & Gerontology; TBK1; PHOSPHORYLATION; ACTIVATION; AUTOPHAGY; WES; Neurosciences; ALS; AMYOTROPHIC-LATERAL-SCLEROSIS; FRONTOTEMPORAL DEMENTIA; GENES; BINDING; Familial ALS; Science & Technology
• Research Categories: Biology, Neuroscience

Abstract

Mutations in TANK binding kinase 1 (TBK1) have been linked to amyotrophic lateral sclerosis. Some TBK1 variants are nonsense and are predicted to cause disease through haploinsufficiency; however, many other mutations are missense with unknown functional effects. We exome sequenced 699 familial amyotrophic lateral sclerosis patients and identified 16 TBK1 novel or extremely rare protein-changing variants. We characterized a subset of these: p.G217R, p.R357X, and p.C471Y. Here, we show that the p.R357X and p.G217R both abolish the ability of TBK1 to phosphorylate 2 of its kinase targets, IRF3 and optineurin, and to undergo phosphorylation. They both inhibit binding to optineurin and the p.G217R, within the TBK1 kinase domain, reduces homodimerization, essential for TBK1 activation and function. Finally, we show that the proportion of TBK1 that is active (phosphorylated) is reduced in 5 lymphoblastoid cell lines derived from patients harboring heterozygous missense or in-frame deletion TBK1 mutations. We conclude that missense mutations in functional domains of TBK1 impair the binding and phosphorylation of its normal targets, implicating a common loss of function mechanism, analogous to truncation mutations.