title: A genome-wide imaging-based screening to identify genes involved in synphilin-1 inclusion formation in Saccharomyces cerevisiae.
abstract: Synphilin-1 is a major component of Parkinson's disease (PD) inclusion bodies implicated in PD pathogenesis. However, the machinery controlling synphilin-1 inclusion formation remains unclear. Here, we investigated synphilin-1 inclusion formation using a systematic genome-wide, high-content imaging based screening approach (HCI) in the yeast Saccharomyces cerevisiae. By combining with a secondary screening for mutants showing significant changes on fluorescence signal intensity, we filtered out hits that significantly decreased the expression level of synphilin-1. We found 133 yeast genes that didn't affect synphilin-1 expression but that were required for the formation of synphilin-1 inclusions. Functional enrichment and physical interaction network analysis revealed these genes to encode for functions involved in cytoskeleton organization, histone modification, sister chromatid segregation, glycolipid biosynthetic process, DNA repair and replication. All hits were confirmed by conventional microscopy. Complementation assays were performed with a selected group of mutants, results indicated that the observed phenotypic changes in synphilin-1 inclusion formation were directly caused by the loss of corresponding genes of the deletion mutants. Further growth assays of these mutants showed a significant synthetic sick effect upon synphilin-1 expression, which supports the hypothesis that matured inclusions represent an end stage of several events meant to protect cells against the synphilin-1 cytotoxicity.
authors.affiliation: School of Life Science and Technology, Harbin Institute of Technology, Harbin 150001, China.
authors.forename: Lei
authors.initials: L
authors.lastname: Zhao
authors.affiliation: School of Life Science and Technology, Harbin Institute of Technology, Harbin 150001, China.
authors.forename: Qian
authors.initials: Q
authors.lastname: Yang
authors.affiliation: Department of Biology, Functional Biology, KU Leuven, 3001 Heverlee, Belgium.
authors.forename: Ju
authors.initials: J
authors.lastname: Zheng
authors.affiliation: Department of Medical Biochemistry and Cell Biology, University of Gothenburg, S-405 30, Göteborg, Sweden.
authors.forename: Xuefeng
authors.initials: X
authors.lastname: Zhu
authors.affiliation: Department of Chemistry and Molecular Biology, University of Gothenburg, S-413 90, Göteborg, Sweden.
authors.forename: Xinxin
authors.initials: X
authors.lastname: Hao
authors.affiliation: School of Life Science and Technology, Harbin Institute of Technology, Harbin 150001, China.
authors.forename: Jia
authors.initials: J
authors.lastname: Song
authors.affiliation: Department of Biology, Functional Biology, KU Leuven, 3001 Heverlee, Belgium.
authors.forename: Tom
authors.initials: T
authors.lastname: Lebacq
authors.affiliation: Department of Biology, Functional Biology, KU Leuven, 3001 Heverlee, Belgium.
authors.forename: Vanessa
authors.initials: V
authors.lastname: Franssens
authors.affiliation: Department of Biology, Functional Biology, KU Leuven, 3001 Heverlee, Belgium.
authors.forename: Joris
authors.initials: J
authors.lastname: Winderickx
authors.affiliation: Department of Chemistry and Molecular Biology, University of Gothenburg, S-413 90, Göteborg, Sweden.
authors.forename: Thomas
authors.initials: T
authors.lastname: Nystrom
authors.affiliation: School of Life Science and Technology, Harbin Institute of Technology, Harbin 150001, China.|Department of Chemistry and Molecular Biology, University of Gothenburg, S-413 90, Göteborg, Sweden.
authors.forename: Beidong
authors.initials: B
authors.lastname: Liu
mesh_terms: D002352:Carrier Proteins / Q000235:genetics* / Q000378:metabolism; D005800:Genes, Fungal; D002479:Inclusion Bodies / Q000378:metabolism*; D012441:Saccharomyces cerevisiae / Q000235:genetics* / Q000378:metabolism
chemical_list: D002352:Carrier Proteins