title: Phase Separation of C9orf72 Dipeptide Repeats Perturbs Stress Granule Dynamics.
abstract: Liquid-liquid phase separation (LLPS) of RNA-binding proteins plays an important role in the formation of multiple membrane-less organelles involved in RNA metabolism, including stress granules. Defects in stress granule homeostasis constitute a cornerstone of ALS/FTLD pathogenesis. Polar residues (tyrosine and glutamine) have been previously demonstrated to be critical for phase separation of ALS-linked stress granule proteins. We now identify an active role for arginine-rich domains in these phase separations. Moreover, arginine-rich dipeptide repeats (DPRs) derived from C9orf72 hexanucleotide repeat expansions similarly undergo LLPS and induce phase separation of a large set of proteins involved in RNA and stress granule metabolism. Expression of arginine-rich DPRs in cells induced spontaneous stress granule assembly that required both eIF2α phosphorylation and G3BP. Together with recent reports showing that DPRs affect nucleocytoplasmic transport, our results point to an important role for arginine-rich DPRs in the pathogenesis of C9orf72 ALS/FTLD.
authors.affiliation: Experimental Neurology and Leuven Research Institute for Neuroscience and Disease (LIND), Department of Neurosciences, KU Leuven - University of Leuven, 3000 Leuven, Belgium; Laboratory of Neurobiology, VIB, Center for Brain and Disease Research, 3000 Leuven, Belgium.
authors.forename: Steven
authors.initials: S
authors.lastname: Boeynaems
authors.affiliation: Experimental Neurology and Leuven Research Institute for Neuroscience and Disease (LIND), Department of Neurosciences, KU Leuven - University of Leuven, 3000 Leuven, Belgium; Laboratory of Neurobiology, VIB, Center for Brain and Disease Research, 3000 Leuven, Belgium.
authors.forename: Elke
authors.initials: E
authors.lastname: Bogaert
authors.affiliation: Center for Structural Biology (CSB), VIB, Vrije Universiteit Brussel (VUB), 1050 Brussels, Belgium.
authors.forename: Denes
authors.initials: D
authors.lastname: Kovacs
authors.affiliation: Biomolecular and Analytical Mass Spectrometry Group, Department of Chemistry, University of Antwerp, 2020 Antwerp, Belgium.
authors.forename: Albert
authors.initials: A
authors.lastname: Konijnenberg
authors.affiliation: VIB-UGent Center for Medical Biotechnology, 9000 Gent, Belgium; VIB Proteomics Core, 9000 Gent, Belgium; Department of Biochemistry, Ghent University, 9000 Gent, Belgium.
authors.forename: Evy
authors.initials: E
authors.lastname: Timmerman
authors.affiliation: Center for Structural Biology (CSB), VIB, Vrije Universiteit Brussel (VUB), 1050 Brussels, Belgium.
authors.forename: Alex
authors.initials: A
authors.lastname: Volkov
authors.affiliation: Center for Structural Biology (CSB), VIB, Vrije Universiteit Brussel (VUB), 1050 Brussels, Belgium.
authors.forename: Mainak
authors.initials: M
authors.lastname: Guharoy
authors.affiliation: Experimental Neurology and Leuven Research Institute for Neuroscience and Disease (LIND), Department of Neurosciences, KU Leuven - University of Leuven, 3000 Leuven, Belgium; Laboratory of Neurobiology, VIB, Center for Brain and Disease Research, 3000 Leuven, Belgium.
authors.forename: Mathias
authors.initials: M
authors.lastname: De Decker
authors.affiliation: Experimental Neurology and Leuven Research Institute for Neuroscience and Disease (LIND), Department of Neurosciences, KU Leuven - University of Leuven, 3000 Leuven, Belgium; Laboratory of Neurobiology, VIB, Center for Brain and Disease Research, 3000 Leuven, Belgium.
authors.forename: Tom
authors.initials: T
authors.lastname: Jaspers
authors.affiliation: Neuroscience Graduate Program, Brown University, Providence, RI 02912, USA.
authors.forename: Veronica H
authors.initials: VH
authors.lastname: Ryan
authors.affiliation: Department of Molecular Pharmacology, Physiology, and Biotechnology, Brown University, Providence, RI 02912, USA.
authors.forename: Abigail M
authors.initials: AM
authors.lastname: Janke
authors.affiliation: VIB, Bioimaging Core Facility, 3000 Leuven, Belgium.
authors.forename: Pieter
authors.initials: P
authors.lastname: Baatsen
authors.affiliation: Laboratory of Virology and Chemotherapy, Rega Institute, KU Leuven, 3000 Leuven, Belgium.
authors.forename: Thomas
authors.initials: T
authors.lastname: Vercruysse
authors.affiliation: Howard Hughes Medical Institute, Department of Cell and Molecular Biology, St. Jude Children's Research Hospital, Memphis, TN 38105, USA.
authors.forename: Regina-Maria
authors.initials: RM
authors.lastname: Kolaitis
authors.affiliation: Laboratory of Virology and Chemotherapy, Rega Institute, KU Leuven, 3000 Leuven, Belgium.
authors.forename: Dirk
authors.initials: D
authors.lastname: Daelemans
authors.affiliation: Howard Hughes Medical Institute, Department of Cell and Molecular Biology, St. Jude Children's Research Hospital, Memphis, TN 38105, USA.
authors.forename: J Paul
authors.initials: JP
authors.lastname: Taylor
authors.affiliation: Division of Rheumatology, Immunology, and Allergy, Harvard Medical School and Brigham and Women's Hospital, Boston, MA 02115, USA.
authors.forename: Nancy
authors.initials: N
authors.lastname: Kedersha
authors.affiliation: Division of Rheumatology, Immunology, and Allergy, Harvard Medical School and Brigham and Women's Hospital, Boston, MA 02115, USA.
authors.forename: Paul
authors.initials: P
authors.lastname: Anderson
authors.affiliation: VIB-UGent Center for Medical Biotechnology, 9000 Gent, Belgium; VIB Proteomics Core, 9000 Gent, Belgium; Department of Biochemistry, Ghent University, 9000 Gent, Belgium.
authors.forename: Francis
authors.initials: F
authors.lastname: Impens
authors.affiliation: Biomolecular and Analytical Mass Spectrometry Group, Department of Chemistry, University of Antwerp, 2020 Antwerp, Belgium; Astbury Centre for Structural Molecular Biology, University of Leeds, Leeds LS2 9JT, UK; School of Molecular and Cellular Biology, University of Leeds, Leeds LS2 9JT, UK.
authors.forename: Frank
authors.initials: F
authors.lastname: Sobott
authors.affiliation: Switch Laboratory, Center for Brain and Disease Research, VIB, 3000 Leuven, Belgium; Switch Laboratory, Department of Cellular and Molecular Medicine, KU Leuven, 3000 Leuven, Belgium.
authors.forename: Joost
authors.initials: J
authors.lastname: Schymkowitz
authors.affiliation: Switch Laboratory, Center for Brain and Disease Research, VIB, 3000 Leuven, Belgium; Switch Laboratory, Department of Cellular and Molecular Medicine, KU Leuven, 3000 Leuven, Belgium.
authors.forename: Frederic
authors.initials: F
authors.lastname: Rousseau
authors.affiliation: Department of Molecular Pharmacology, Physiology, and Biotechnology, Brown University, Providence, RI 02912, USA.
authors.forename: Nicolas L
authors.initials: NL
authors.lastname: Fawzi
authors.affiliation: Experimental Neurology and Leuven Research Institute for Neuroscience and Disease (LIND), Department of Neurosciences, KU Leuven - University of Leuven, 3000 Leuven, Belgium; Department of Neurology, University Hospitals Leuven, 3000 Leuven, Belgium.
authors.forename: Wim
authors.initials: W
authors.lastname: Robberecht
authors.affiliation: Experimental Neurology and Leuven Research Institute for Neuroscience and Disease (LIND), Department of Neurosciences, KU Leuven - University of Leuven, 3000 Leuven, Belgium; Laboratory of Neurobiology, VIB, Center for Brain and Disease Research, 3000 Leuven, Belgium; Department of Neurology, University Hospitals Leuven, 3000 Leuven, Belgium.
authors.forename: Philip
authors.initials: P
authors.lastname: Van Damme
authors.affiliation: Center for Structural Biology (CSB), VIB, Vrije Universiteit Brussel (VUB), 1050 Brussels, Belgium; Institute of Enzymology, Research Centre for Natural Sciences of the Hungarian Academy of Sciences, 1117 Budapest, Hungary. Electronic address: ptompa@vub.ac.be.
authors.forename: Peter
authors.initials: P
authors.lastname: Tompa
authors.affiliation: Experimental Neurology and Leuven Research Institute for Neuroscience and Disease (LIND), Department of Neurosciences, KU Leuven - University of Leuven, 3000 Leuven, Belgium; Laboratory of Neurobiology, VIB, Center for Brain and Disease Research, 3000 Leuven, Belgium. Electronic address: ludo.vandenbosch@vib-kuleuven.be.
authors.forename: Ludo
authors.initials: L
authors.lastname: Van Den Bosch
mesh_terms: D000690:Amyotrophic Lateral Sclerosis / Q000235:genetics / Q000378:metabolism* / Q000473:pathology; D001120:Arginine / Q000737:chemistry / Q000378:metabolism*; D000073885:C9orf72 Protein; D002352:Carrier Proteins / Q000235:genetics / Q000378:metabolism; D003594:Cytoplasmic Granules / Q000378:metabolism* / Q000473:pathology; D004265:DNA Helicases; D004151:Dipeptides / Q000737:chemistry / Q000378:metabolism*; D015852:Eukaryotic Initiation Factor-2 / Q000235:genetics / Q000378:metabolism; D006367:HeLa Cells; D006801:Humans; D064267:Intrinsically Disordered Proteins / Q000737:chemistry / Q000378:metabolism*; D066292:Lipid Droplets / Q000378:metabolism; D010766:Phosphorylation; D000075223:Poly-ADP-Ribose Binding Proteins; D000072417:Protein Domains; D011506:Proteins / Q000737:chemistry / Q000378:metabolism*; D012313:RNA / Q000378:metabolism; D020365:RNA Helicases; D000067776:RNA Recognition Motif Proteins; D013997:Time Factors; D014162:Transfection
chemical_list: D000073885:C9orf72 Protein; C568651:C9orf72 protein, human; D002352:Carrier Proteins; D004151:Dipeptides; D015852:Eukaryotic Initiation Factor-2; D064267:Intrinsically Disordered Proteins; D000075223:Poly-ADP-Ribose Binding Proteins; D011506:Proteins; D000067776:RNA Recognition Motif Proteins; D012313:RNA; D001120:Arginine; D004265:DNA Helicases; C116070:G3BP1 protein, human; D020365:RNA Helicases
keywords: FUS; LLPS; amyotrophic lateral sclerosis; frontotemporal lobar degeneration; hnRNP; intrinsically disordered protein; low complexity domain; phase transition; prion-like domain; protein aggregation