title: Identification of novel regulators of apoptosis using a high-throughput cell-based screen.
abstract: High-throughput subcellular imaging is a powerful tool for investigating the function of genes. In order to identify novel regulators of apoptosis we transiently transfected HeLa cells with 938 hypothetical genes of unknown function, and captured their nuclear images with an automated fluorescence microscope. We selected genes that induced greater than 3-fold increase in the percentage of apoptotic nuclei compared with vector-transfected cells. The full-length genes C10orf61, MGC 26717, and FLJ13855 were identified as candidate proapoptotic genes, and their apoptotic effects were confirmed by DNA fragmentation ELISAs and Western blotting for caspase-7 and PARP. We conclude that a subcellular image-based apoptotic screen is useful for identifying genes with proapoptotic activity.
authors.affiliation: Functional Genomics Research Center, Korea Research Institute of Bioscience and Biotechnology, Daejeon 305-806, Korea.
authors.forename: Kyung Mi
authors.initials: KM
authors.lastname: Park
authors.forename: Eunju
authors.initials: E
authors.lastname: Kang
authors.forename: Yeo-Jin
authors.initials: YJ
authors.lastname: Jeon
authors.forename: Nayoung
authors.initials: N
authors.lastname: Kim
authors.forename: Nam-Soon
authors.initials: NS
authors.lastname: Kim
authors.forename: Hyang-Sook
authors.initials: HS
authors.lastname: Yoo
authors.forename: Young Il
authors.initials: YI
authors.lastname: Yeom
authors.forename: Soo Jung
authors.initials: SJ
authors.lastname: Kim
mesh_terms: D017209:Apoptosis / Q000502:physiology*; D051017:Apoptosis Regulatory Proteins / Q000378:metabolism*; D053179:Caspase 7 / Q000378:metabolism; D024161:Collagen Type XI / Q000378:metabolism; D053938:DNA Fragmentation; D006367:HeLa Cells; D006801:Humans; D007091:Image Processing, Computer-Assisted / Q000379:methods; D008856:Microscopy, Fluorescence; D010957:Plasmids / Q000378:metabolism
chemical_list: D051017:Apoptosis Regulatory Proteins; C499319:COL11A2 protein, human; D024161:Collagen Type XI; D053179:Caspase 7