TDP-43 is a ubiquitylation substrate of the SCF cyclin F complex.

Autor: Rayner SL; Centre for Motor Neuron Disease Research, Department of Biomedical Sciences, Faculty of Medicine, Health, and Human Sciences, Macquarie University, North Ryde, NSW, Australia. Electronic address: stephanie.rayner@mq.edu.au., Yang S; Centre for Motor Neuron Disease Research, Department of Biomedical Sciences, Faculty of Medicine, Health, and Human Sciences, Macquarie University, North Ryde, NSW, Australia. Electronic address: shu.yang@mq.edu.au., Farrawell NE; Illawarra Health and Medical Research Institute (IHMRI), University of Wollongong, Wollongong, NSW, Australia; School of Chemistry and Molecular Bioscience and Molecular Horizons, University of Wollongong, Wollongong, NSW, Australia. Electronic address: nfarrawe@uow.edu.au., Jagaraj CJ; Centre for Motor Neuron Disease Research, Department of Biomedical Sciences, Faculty of Medicine, Health, and Human Sciences, Macquarie University, North Ryde, NSW, Australia. Electronic address: cyril-jones.jagaraj@mq.edu.au., Cheng F; Centre for Motor Neuron Disease Research, Department of Biomedical Sciences, Faculty of Medicine, Health, and Human Sciences, Macquarie University, North Ryde, NSW, Australia. Electronic address: flora.cheng@mq.edu.au., Davidson JM; Centre for Motor Neuron Disease Research, Department of Biomedical Sciences, Faculty of Medicine, Health, and Human Sciences, Macquarie University, North Ryde, NSW, Australia. Electronic address: jennilee.davidson@hdr.mq.edu.au., Luu L; Centre for Motor Neuron Disease Research, Department of Biomedical Sciences, Faculty of Medicine, Health, and Human Sciences, Macquarie University, North Ryde, NSW, Australia. Electronic address: luan.luu@mq.edu.au., Redondo AG; Unidad de ELA, Instituto de Investigación Hospital 12 de Octubre de Madrid, SERMAS, Centro de Investigación Biomédica en Red de Enfermedades Raras (CIBERER U-723), Madrid, Spain. Electronic address: ela@correo.h12o.es., Rábano A; Neuropathology Department and CIEN Tissue Bank, Alzheimer's Centre Reina Sofia-CIEN Foundation, 28031 Madrid, Spain. Electronic address: arabano@fundacioncien.es., Borrego-Hernández D; Unidad de ELA, Instituto de Investigación Hospital 12 de Octubre de Madrid, SERMAS, Centro de Investigación Biomédica en Red de Enfermedades Raras (CIBERER U-723), Madrid, Spain. Electronic address: dborregohernandez.imas12@h12o.es., Atkin JD; Centre for Motor Neuron Disease Research, Department of Biomedical Sciences, Faculty of Medicine, Health, and Human Sciences, Macquarie University, North Ryde, NSW, Australia. Electronic address: julie.atkin@mq.edu.au., Morsch M; Centre for Motor Neuron Disease Research, Department of Biomedical Sciences, Faculty of Medicine, Health, and Human Sciences, Macquarie University, North Ryde, NSW, Australia. Electronic address: marco.morsch@mq.edu.au., Blair IP; Centre for Motor Neuron Disease Research, Department of Biomedical Sciences, Faculty of Medicine, Health, and Human Sciences, Macquarie University, North Ryde, NSW, Australia. Electronic address: ian.blair@mq.edu.au., Yerbury JJ; Illawarra Health and Medical Research Institute (IHMRI), University of Wollongong, Wollongong, NSW, Australia; School of Chemistry and Molecular Bioscience and Molecular Horizons, University of Wollongong, Wollongong, NSW, Australia. Electronic address: jyerbury@uow.edu.au., Chung R; Centre for Motor Neuron Disease Research, Department of Biomedical Sciences, Faculty of Medicine, Health, and Human Sciences, Macquarie University, North Ryde, NSW, Australia. Electronic address: roger.chung@mq.edu.au., Lee A; Centre for Motor Neuron Disease Research, Department of Biomedical Sciences, Faculty of Medicine, Health, and Human Sciences, Macquarie University, North Ryde, NSW, Australia. Electronic address: albert.lee@mq.edu.au.
Jazyk: angličtina
Zdroj: Neurobiology of disease [Neurobiol Dis] 2022 Jun 01; Vol. 167, pp. 105673. Date of Electronic Publication: 2022 Feb 26.
DOI: 10.1016/j.nbd.2022.105673
Abstrakt: Background: Amyotrophic lateral sclerosis (ALS) is a fatal neurodegenerative disease characterised by the loss of upper and lower motor neurons in the brain and spinal cord. ALS and frontotemporal dementia (FTD) are overlapping diseases with shared pathological features. Affected neurons of people with ALS and FTD typically contain ubiquitin-immunoreactive inclusions, of which TDP-43 (Tar DNA-binding protein of 43 kDa) is a major component. However, what triggers the formation of these abnormal TDP-43 inclusions is unclear. Previously, we identified CCNF mutations in cohorts of familial and sporadic cases of ALS and FTD. CCNF encodes cyclin F, the substrate-binding component of a multiprotein E3 ubiquitin ligase complex that ubiquitylates and subsequently directs a set of protein substrates for proteasomal degradation. Here, we explored the relationship between cyclin F and TDP-43.
Methods: We used a series of complementary biochemical approaches including immunoprecipitations, in vitro ubiquitylation assays, immunofluorescence imaging and immunocytochemistry. Unpaired student t-tests were used to determine statistical significance of the results.
Results: In this study, we demonstrate that that the SCF cyclin F complex directly mediates the poly-ubiquitylation of TDP-43. Importantly, we demonstrate that cyclin F bearing the pathogenic ALS/FTD mutation, S621G, leads to aberrant ubiquitylation of TDP-43 as well as the accumulation of K48-ubiquitylated TDP-43 in neuron-like cells. Furthermore, we demonstrate that a patient carrying the ALS/FTD cyclin F S195R mutation displayed skein-like cytoplasmic TDP-43 aggregates, implying abnormal TDP-43 degradation in a CCNF mutation bearing patient.
Conclusion: In summary, this study reports a direct ubiquitylation mechanism for TDP-43, revealing important insights into the regulation of cyclin F-mediated TDP-43 turnover and clues towards understanding the molecular origins of the ubiquitylated TDP-43 inclusions that are the hallmark pathological feature in ALS and FTD.
(Copyright © 2022 The Authors. Published by Elsevier Inc. All rights reserved.)
Databáze: MEDLINE
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