title: Novel suppressors of alpha-synuclein toxicity identified using yeast.
abstract: The mechanism by which the Parkinson's disease-related protein alpha-synuclein (alpha-syn) causes neurodegeneration has not been elucidated. To determine the genes that protect cells from alpha-syn, we used a genetic screen to identify suppressors of the super sensitivity of the yeast Saccharomyces cerevisiae expressing alpha-syn to killing by hydrogen peroxide. Forty genes in ubiquitin-dependent protein catabolism, protein biosynthesis, vesicle trafficking and the response to stress were identified. Five of the forty genes--ENT3, IDP3, JEM1, ARG2 and HSP82--ranked highest in their ability to block alpha-syn-induced reactive oxygen species accumulation, and these five genes were characterized in more detail. The deletion of any of these five genes enhanced the toxicity of alpha-syn as judged by growth defects compared with wild-type cells expressing alpha-syn, which indicates that these genes protect cells from alpha-syn. Strikingly, four of the five genes are specific for alpha-syn in that they fail to protect cells from the toxicity of the two inherited mutants A30P or A53T. This finding suggests that alpha-syn causes toxicity to cells through a different pathway than these two inherited mutants. Lastly, overexpression of Ent3p, which is a clathrin adapter protein involved in protein transport between the Golgi and the vacuole, causes alpha-syn to redistribute from the plasma membrane into cytoplasmic vesicular structures. Our interpretation is that Ent3p mediates the transport of alpha-syn to the vacuole for proteolytic degradation. A similar clathrin adaptor protein, epsinR, exists in humans.
authors.affiliation: Department of Biochemistry and Molecular Biology, Louisiana State University Health Sciences Center, Shreveport, LA 71130-3932, USA.
authors.forename: Jun
authors.initials: J
authors.lastname: Liang
authors.forename: Cheryl
authors.initials: C
authors.lastname: Clark-Dixon
authors.forename: Shaoxiao
authors.initials: S
authors.lastname: Wang
authors.forename: Todd R
authors.initials: TR
authors.lastname: Flower
authors.forename: Tara
authors.initials: T
authors.lastname: Williams-Hart
authors.forename: Richard
authors.initials: R
authors.lastname: Zweig
authors.forename: Lucy C
authors.initials: LC
authors.lastname: Robinson
authors.forename: Kelly
authors.initials: K
authors.lastname: Tatchell
authors.forename: Stephan N
authors.initials: SN
authors.lastname: Witt
mesh_terms: D033942:Adaptor Proteins, Vesicular Transport / Q000235:genetics / Q000378:metabolism; D018841:HSP90 Heat-Shock Proteins / Q000235:genetics / Q000378:metabolism; D006801:Humans; D007521:Isocitrate Dehydrogenase / Q000235:genetics / Q000378:metabolism; D008565:Membrane Proteins / Q000235:genetics / Q000378:metabolism; D018832:Molecular Chaperones; D010300:Parkinson Disease / Q000235:genetics* / Q000378:metabolism; D021381:Protein Transport; D017382:Reactive Oxygen Species / Q000378:metabolism; D012441:Saccharomyces cerevisiae / Q000187:drug effects / Q000235:genetics* / Q000378:metabolism; D029701:Saccharomyces cerevisiae Proteins / Q000235:genetics / Q000378:metabolism; D013489:Suppression, Genetic; D051844:alpha-Synuclein / Q000235:genetics / Q000378:metabolism / Q000633:toxicity*
chemical_list: D033942:Adaptor Proteins, Vesicular Transport; C495587:Ent3 protein, S cerevisiae; C074510:HSP82 protein, S cerevisiae; D018841:HSP90 Heat-Shock Proteins; C106168:JEM1 protein, S cerevisiae; D008565:Membrane Proteins; D018832:Molecular Chaperones; D017382:Reactive Oxygen Species; D029701:Saccharomyces cerevisiae Proteins; D051844:alpha-Synuclein; D007521:Isocitrate Dehydrogenase; C039355:isocitrate dehydrogenase (NADP+)
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