title: A yeast model of FUS/TLS-dependent cytotoxicity.
abstract: FUS/TLS is a nucleic acid binding protein that, when mutated, can cause a subset of familial amyotrophic lateral sclerosis (fALS). Although FUS/TLS is normally located predominantly in the nucleus, the pathogenic mutant forms of FUS/TLS traffic to, and form inclusions in, the cytoplasm of affected spinal motor neurons or glia. Here we report a yeast model of human FUS/TLS expression that recapitulates multiple salient features of the pathology of the disease-causing mutant proteins, including nuclear to cytoplasmic translocation, inclusion formation, and cytotoxicity. Protein domain analysis indicates that the carboxyl-terminus of FUS/TLS, where most of the ALS-associated mutations are clustered, is required but not sufficient for the toxicity of the protein. A genome-wide genetic screen using a yeast over-expression library identified five yeast DNA/RNA binding proteins, encoded by the yeast genes ECM32, NAM8, SBP1, SKO1, and VHR1, that rescue the toxicity of human FUS/TLS without changing its expression level, cytoplasmic translocation, or inclusion formation. Furthermore, hUPF1, a human homologue of ECM32, also rescues the toxicity of FUS/TLS in this model, validating the yeast model and implicating a possible insufficiency in RNA processing or the RNA quality control machinery in the mechanism of FUS/TLS mediated toxicity. Examination of the effect of FUS/TLS expression on the decay of selected mRNAs in yeast indicates that the nonsense-mediated decay pathway is probably not the major determinant of either toxicity or suppression.
authors.affiliation: Department of Biochemistry and Chemistry, Rosenstiel Basic Medical Sciences Research Center, Brandeis University, Waltham, Massachusetts, United States of America.
authors.forename: Shulin
authors.initials: S
authors.lastname: Ju
authors.forename: Daniel F
authors.initials: DF
authors.lastname: Tardiff
authors.forename: Haesun
authors.initials: H
authors.lastname: Han
authors.forename: Kanneganti
authors.initials: K
authors.lastname: Divya
authors.forename: Quan
authors.initials: Q
authors.lastname: Zhong
authors.forename: Lynne E
authors.initials: LE
authors.lastname: Maquat
authors.forename: Daryl A
authors.initials: DA
authors.lastname: Bosco
authors.forename: Lawrence J
authors.initials: LJ
authors.lastname: Hayward
authors.forename: Robert H
authors.initials: RH
authors.lastname: Brown
authors.forename: Susan
authors.initials: S
authors.lastname: Lindquist
authors.forename: Dagmar
authors.initials: D
authors.lastname: Ringe
authors.forename: Gregory A
authors.initials: GA
authors.lastname: Petsko
mesh_terms: D000690:Amyotrophic Lateral Sclerosis / Q000235:genetics / Q000473:pathology; D002467:Cell Nucleus / Q000235:genetics; D003593:Cytoplasm / Q000235:genetics / Q000378:metabolism; D004265:DNA Helicases / Q000235:genetics / Q000378:metabolism*; D005786:Gene Expression Regulation; D009154:Mutation; D009474:Neurons / Q000378:metabolism; D020365:RNA Helicases; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D034702:RNA-Binding Protein FUS / Q000235:genetics* / Q000378:metabolism; D011993:Recombinant Fusion Proteins / Q000378:metabolism; D012441:Saccharomyces cerevisiae / Q000235:genetics* / Q000378:metabolism; D029701:Saccharomyces cerevisiae Proteins / Q000235:genetics / Q000378:metabolism*; D015534:Trans-Activators / Q000378:metabolism*
chemical_list: D012333:RNA, Messenger; D034702:RNA-Binding Protein FUS; D011993:Recombinant Fusion Proteins; D029701:Saccharomyces cerevisiae Proteins; D015534:Trans-Activators; D004265:DNA Helicases; D020365:RNA Helicases; C101828:UPF1 protein, human; C091576:ECM32 protein, S cerevisiae