title: Monomethylated and unmethylated FUS exhibit increased binding to Transportin and distinguish FTLD-FUS from ALS-FUS.
abstract: Deposition of the nuclear DNA/RNA-binding protein Fused in sarcoma (FUS) in cytosolic inclusions is a common hallmark of some cases of frontotemporal lobar degeneration (FTLD-FUS) and amyotrophic lateral sclerosis (ALS-FUS). Whether both diseases also share common pathological mechanisms is currently unclear. Based on our previous finding that FUS deposits are hypomethylated in FTLD-FUS but not in ALS-FUS, we have now investigated whether genetic or pharmacological inactivation of Protein arginine methyltransferase 1 (PRMT1) activity results in unmethylated FUS or in alternatively methylated forms of FUS. To do so, we generated FUS-specific monoclonal antibodies that specifically recognize unmethylated arginine (UMA), monomethylated arginine (MMA) or asymmetrically dimethylated arginine (ADMA). Loss of PRMT1 indeed not only results in an increase of UMA FUS and a decrease of ADMA FUS, but also in a significant increase of MMA FUS. Compared to ADMA FUS, UMA and MMA FUS exhibit much higher binding affinities to Transportin-1, the nuclear import receptor of FUS, as measured by pull-down assays and isothermal titration calorimetry. Moreover, we show that MMA FUS occurs exclusively in FTLD-FUS, but not in ALS-FUS. Our findings therefore provide additional evidence that FTLD-FUS and ALS-FUS are caused by distinct disease mechanisms although both share FUS deposits as a common denominator.
authors.affiliation: Biomedical Center (BMC), Biochemistry, Ludwig-Maximilians-University Munich, Feodor-Lynen Strasse 17, 81377, Munich, Germany.|German Center for Neurodegenerative Diseases (DZNE) Munich, Feodor-Lynen Strasse 17, 81377, Munich, Germany.|Universitat Autònoma de Barcelona, 08193, Bellaterra, Barcelona, Spain.
authors.forename: Marc
authors.initials: M
authors.lastname: Suárez-Calvet
authors.affiliation: Department of Neuropathology, University of Tübingen, 72076, Tübingen, Germany.|DZNE, German Center for Neurodegenerative Diseases, 72076, Tübingen, Germany.
authors.forename: Manuela
authors.initials: M
authors.lastname: Neumann
authors.affiliation: Department of Psychiatry and Psychotherapy, Ludwig-Maximilians-University Munich, 80336, Munich, Germany.|Center for Neuropathology and Prion Research, Ludwig-Maximilians-University Munich, 81377, Munich, Germany.|German Center for Neurodegenerative Diseases (DZNE) Munich, Feodor-Lynen Strasse 17, 81377, Munich, Germany.
authors.forename: Thomas
authors.initials: T
authors.lastname: Arzberger
authors.affiliation: Biomedical Center (BMC), Biochemistry, Ludwig-Maximilians-University Munich, Feodor-Lynen Strasse 17, 81377, Munich, Germany.|BioMedical Center (BMC), Lehrstuhl Zellbiologie (Anatomie III), Großhaderner Strasse 9, 82152, Planegg-Martinsried, Germany.
authors.forename: Claudia
authors.initials: C
authors.lastname: Abou-Ajram
authors.affiliation: Biomedical Center (BMC), Biochemistry, Ludwig-Maximilians-University Munich, Feodor-Lynen Strasse 17, 81377, Munich, Germany.
authors.forename: Eva
authors.initials: E
authors.lastname: Funk
authors.affiliation: German Center for Neurodegenerative Diseases (DZNE) Munich, Feodor-Lynen Strasse 17, 81377, Munich, Germany.|Munich Cluster for Systems Neurology (SyNergy), 81377, Munich, Germany.
authors.forename: Hannelore
authors.initials: H
authors.lastname: Hartmann
authors.affiliation: German Center for Neurodegenerative Diseases (DZNE) Munich, Feodor-Lynen Strasse 17, 81377, Munich, Germany.|Munich Cluster for Systems Neurology (SyNergy), 81377, Munich, Germany.
authors.forename: Dieter
authors.initials: D
authors.lastname: Edbauer
authors.affiliation: Institute of Molecular Immunology, Helmholtz Zentrum München, German Research Center for Environmental Health (GmbH), 81377, Munich, Germany.
authors.forename: Elisabeth
authors.initials: E
authors.lastname: Kremmer
authors.affiliation: Department of Chemistry, Center for Integrated Protein Science Munich (CIPSM), Technische Universität München, Lichtenbergstr.4, 85747, Garching, Germany.|Institute of Structural Biology, Helmholtz Zentrum München, 85764, Neuherberg, Germany.
authors.forename: Christoph
authors.initials: C
authors.lastname: Göbl
authors.affiliation: Department of Chemistry, Center for Integrated Protein Science Munich (CIPSM), Technische Universität München, Lichtenbergstr.4, 85747, Garching, Germany.|Institute of Structural Biology, Helmholtz Zentrum München, 85764, Neuherberg, Germany.
authors.forename: Moritz
authors.initials: M
authors.lastname: Resch
authors.affiliation: Department of Chemistry, Center for Integrated Protein Science Munich (CIPSM), Technische Universität München, Lichtenbergstr.4, 85747, Garching, Germany.|Institute of Structural Biology, Helmholtz Zentrum München, 85764, Neuherberg, Germany.
authors.forename: Benjamin
authors.initials: B
authors.lastname: Bourgeois
authors.affiliation: Department of Chemistry, Center for Integrated Protein Science Munich (CIPSM), Technische Universität München, Lichtenbergstr.4, 85747, Garching, Germany.|Institute of Structural Biology, Helmholtz Zentrum München, 85764, Neuherberg, Germany.|Institute of Molecular Biology and Biochemistry, Center of Molecular Medicine, Medical University of Graz, 8010, Graz, Austria.|Omics Center Graz, BioTechMed, 8010, Graz, Austria.
authors.forename: Tobias
authors.initials: T
authors.lastname: Madl
authors.affiliation: Department of Chemistry and Biochemistry, University of Bern, 3012, Bern, Switzerland.|Graduate School for Cellular and Biomedical Sciences, University of Bern, 3012, Bern, Switzerland.
authors.forename: Stefan
authors.initials: S
authors.lastname: Reber
authors.affiliation: Department of Chemistry and Biochemistry, University of Bern, 3012, Bern, Switzerland.
authors.forename: Daniel
authors.initials: D
authors.lastname: Jutzi
authors.affiliation: Department of Chemistry and Biochemistry, University of Bern, 3012, Bern, Switzerland.
authors.forename: Marc-David
authors.initials: MD
authors.lastname: Ruepp
authors.affiliation: Department of Pathology, Vancouver General Hospital, University of British Columbia, Vancouver, Canada.
authors.forename: Ian R A
authors.initials: IR
authors.lastname: Mackenzie
authors.affiliation: Department of Neuropathology, John Radcliffe Hospital, Oxford, UK.
authors.forename: Olaf
authors.initials: O
authors.lastname: Ansorge
authors.affiliation: Biomedical Center (BMC), Biochemistry, Ludwig-Maximilians-University Munich, Feodor-Lynen Strasse 17, 81377, Munich, Germany. dorothee.dormann@med.uni-muenchen.de.|BioMedical Center (BMC), Lehrstuhl Zellbiologie (Anatomie III), Großhaderner Strasse 9, 82152, Planegg-Martinsried, Germany. dorothee.dormann@med.uni-muenchen.de.|Munich Cluster for Systems Neurology (SyNergy), 81377, Munich, Germany. dorothee.dormann@med.uni-muenchen.de.
authors.forename: Dorothee
authors.initials: D
authors.lastname: Dormann
authors.affiliation: Biomedical Center (BMC), Biochemistry, Ludwig-Maximilians-University Munich, Feodor-Lynen Strasse 17, 81377, Munich, Germany. christian.haass@mail03.med.uni-muenchen.de.|German Center for Neurodegenerative Diseases (DZNE) Munich, Feodor-Lynen Strasse 17, 81377, Munich, Germany. christian.haass@mail03.med.uni-muenchen.de.|Munich Cluster for Systems Neurology (SyNergy), 81377, Munich, Germany. christian.haass@mail03.med.uni-muenchen.de.
authors.forename: Christian
authors.initials: C
authors.lastname: Haass
mesh_terms: D000690:Amyotrophic Lateral Sclerosis / Q000235:genetics / Q000378:metabolism*; D000818:Animals; D000906:Antibodies / Q000494:pharmacology; D001120:Arginine / Q000378:metabolism; D002478:Cells, Cultured; D002540:Cerebral Cortex / Q000166:cytology; D004622:Embryo, Mammalian; D053595:Embryonic Stem Cells; D004791:Enzyme Inhibitors / Q000494:pharmacology; D005260:Female; D057174:Frontotemporal Lobar Degeneration / Q000235:genetics / Q000378:metabolism*; D006801:Humans; D002479:Inclusion Bodies / Q000187:drug effects / Q000378:metabolism; D008297:Male; D008745:Methylation; D051379:Mice; D008810:Mice, Inbred C57BL; D018345:Mice, Knockout; D009474:Neurons / Q000187:drug effects / Q000378:metabolism; D011485:Protein Binding / Q000187:drug effects / Q000235:genetics; D011484:Protein-Arginine N-Methyltransferases / Q000235:genetics / Q000378:metabolism*; D034702:RNA-Binding Protein FUS / Q000276:immunology / Q000378:metabolism*; D051381:Rats; D028961:beta Karyopherins / Q000276:immunology / Q000378:metabolism*
chemical_list: D000906:Antibodies; D004791:Enzyme Inhibitors; D034702:RNA-Binding Protein FUS; C108121:TNPO1 protein, human; D028961:beta Karyopherins; D001120:Arginine; D011484:Protein-Arginine N-Methyltransferases
keywords: Amyotrophic lateral sclerosis (ALS); Arginine methylation; Frontotemporal lobar degeneration (FTLD); Fused in sarcoma (FUS); Neurodegeneration; Protein arginine methyltransferase 1 (PRMT1); Transportin-1