title: Mutant human FUS Is ubiquitously mislocalized and generates persistent stress granules in primary cultured transgenic zebrafish cells.
abstract: FUS mutations can occur in familial amyotrophic lateral sclerosis (fALS), a neurodegenerative disease with cytoplasmic FUS inclusion bodies in motor neurons. To investigate FUS pathology, we generated transgenic zebrafish expressing GFP-tagged wild-type or fALS (R521C) human FUS. Cell cultures were made from these zebrafish and the subcellular localization of human FUS and the generation of stress granule (SG) inclusions examined in different cell types, including differentiated motor neurons. We demonstrate that mutant FUS is mislocalized from the nucleus to the cytosol to a similar extent in motor neurons and all other cell types. Both wild-type and R521C FUS localized to SGs in zebrafish cells, demonstrating an intrinsic ability of human FUS to accumulate in SGs irrespective of the presence of disease-associated mutations or specific cell type. However, elevation in relative cytosolic to nuclear FUS by the R521C mutation led to a significant increase in SG assembly and persistence within a sub population of vulnerable cells, although these cells were not selectively motor neurons.
authors.affiliation: The Brain & Mind Research Institute, University of Sydney, Sydney, New South Wales, Australia; The Bosch Institute, University of Sydney, Sydney, New South Wales, Australia; Discipline of Anatomy and Histology, University of Sydney, Sydney, New South Wales, Australia.
authors.forename: Jamie Rae
authors.initials: JR
authors.lastname: Acosta
authors.affiliation: The Brain & Mind Research Institute, University of Sydney, Sydney, New South Wales, Australia; The Bosch Institute, University of Sydney, Sydney, New South Wales, Australia; Discipline of Anatomy and Histology, University of Sydney, Sydney, New South Wales, Australia.
authors.forename: Claire
authors.initials: C
authors.lastname: Goldsbury
authors.affiliation: The Bosch Institute, University of Sydney, Sydney, New South Wales, Australia; Discipline of Anatomy and Histology, University of Sydney, Sydney, New South Wales, Australia; Motorneurone Disease Research Centre, Australian School of Advanced Medicine, Macquarie University, Sydney, New South Wales, Australia.
authors.forename: Claire
authors.initials: C
authors.lastname: Winnick
authors.affiliation: The Bosch Institute, University of Sydney, Sydney, New South Wales, Australia; Discipline of Anatomy and Histology, University of Sydney, Sydney, New South Wales, Australia; Motorneurone Disease Research Centre, Australian School of Advanced Medicine, Macquarie University, Sydney, New South Wales, Australia.
authors.forename: Andrew P
authors.initials: AP
authors.lastname: Badrock
authors.affiliation: The Bosch Institute, University of Sydney, Sydney, New South Wales, Australia; Discipline of Physiology, University of Sydney, Sydney, New South Wales, Australia.
authors.forename: Stuart T
authors.initials: ST
authors.lastname: Fraser
authors.affiliation: The Brain & Mind Research Institute, University of Sydney, Sydney, New South Wales, Australia; ANZAC Research Institute, University of Sydney, Sydney, New South Wales, Australia.
authors.forename: Angela S
authors.initials: AS
authors.lastname: Laird
authors.affiliation: Institute for Molecular Bioscience, The University of Queensland, Brisbane, Australia.
authors.forename: Thomas E
authors.initials: TE
authors.lastname: Hall
authors.affiliation: The Bosch Institute, University of Sydney, Sydney, New South Wales, Australia; Motorneurone Disease Research Centre, Australian School of Advanced Medicine, Macquarie University, Sydney, New South Wales, Australia.
authors.forename: Emily K
authors.initials: EK
authors.lastname: Don
authors.affiliation: ANZAC Research Institute, University of Sydney, Sydney, New South Wales, Australia; Motorneurone Disease Research Centre, Australian School of Advanced Medicine, Macquarie University, Sydney, New South Wales, Australia.
authors.forename: Jennifer A
authors.initials: JA
authors.lastname: Fifita
authors.affiliation: ANZAC Research Institute, University of Sydney, Sydney, New South Wales, Australia; Motorneurone Disease Research Centre, Australian School of Advanced Medicine, Macquarie University, Sydney, New South Wales, Australia.
authors.forename: Ian P
authors.initials: IP
authors.lastname: Blair
authors.affiliation: ANZAC Research Institute, University of Sydney, Sydney, New South Wales, Australia; Motorneurone Disease Research Centre, Australian School of Advanced Medicine, Macquarie University, Sydney, New South Wales, Australia.
authors.forename: Garth A
authors.initials: GA
authors.lastname: Nicholson
authors.affiliation: The Bosch Institute, University of Sydney, Sydney, New South Wales, Australia; Discipline of Anatomy and Histology, University of Sydney, Sydney, New South Wales, Australia; Motorneurone Disease Research Centre, Australian School of Advanced Medicine, Macquarie University, Sydney, New South Wales, Australia.
authors.forename: Nicholas J
authors.initials: NJ
authors.lastname: Cole
mesh_terms: D000818:Animals; D030801:Animals, Genetically Modified; D002478:Cells, Cultured; D003594:Cytoplasmic Granules / Q000378:metabolism*; D003600:Cytosol / Q000378:metabolism*; D006801:Humans; D009046:Motor Neurons / Q000166:cytology; D009154:Mutation; D021381:Protein Transport / Q000235:genetics; D034702:RNA-Binding Protein FUS / Q000235:genetics* / Q000378:metabolism*; D015027:Zebrafish / Q000235:genetics* / Q000378:metabolism*
chemical_list: D034702:RNA-Binding Protein FUS