title: High-content functional genomic screening to identify novel regulators of the PINK1-Parkin pathway.
abstract: PINK1/PARK6 and Parkin/PARK2 are amongst the most commonly mutated genes associated with recessive forms of familial Parkinson's disease. Recent evidence indicates that the proteins they encode, PINK1 and Parkin, function in the same pathway to mediate the selective autophagic clearance of dysfunctional mitochondria. Upon mitochondrial damage, PINK1 is stabilized on the outer mitochondrial membrane where it phosphorylates ubiquitin, generating a signal for the recruitment and activation of Parkin. However, key mechanistic questions still exist regarding Parkin recruitment, including whether or not other factors are required for the PINK1 and Parkin pathway. We describe a method below using high-throughput RNA interference technology to interrogate the genome for novel components of the PINK1 and Parkin pathway.
authors.affiliation: Children's Hospital of Eastern Ontario Research Institute, Ottawa, Ontario, Canada; Department of Biochemistry, Microbiology, and Immunology, University of Ottawa, Ottawa, Ontario, Canada.
authors.forename: Andy Cheuk-Him
authors.initials: AC
authors.lastname: Ng
authors.affiliation: Children's Hospital of Eastern Ontario Research Institute, Ottawa, Ontario, Canada.
authors.forename: Stephen D
authors.initials: SD
authors.lastname: Baird
authors.affiliation: Children's Hospital of Eastern Ontario Research Institute, Ottawa, Ontario, Canada; Department of Biochemistry, Microbiology, and Immunology, University of Ottawa, Ottawa, Ontario, Canada; Department of Cellular and Molecular Medicine, University of Ottawa, Ottawa, Ontario, Canada; Department of Pediatrics, University of Ottawa, Ottawa, Ontario, Canada. Electronic address: rob@arc.cheo.ca.
authors.forename: Robert A
authors.initials: RA
authors.lastname: Screaton
mesh_terms: D000465:Algorithms; D001330:Electronic Data Processing; D023281:Genomics / Q000295:instrumentation / Q000379:methods*; D006367:HeLa Cells; D059014:High-Throughput Nucleotide Sequencing / Q000295:instrumentation / Q000379:methods; D006801:Humans; D011494:Protein Kinases / Q000235:genetics / Q000378:metabolism*; D011506:Proteins / Q000235:genetics / Q000378:metabolism; D034622:RNA Interference; D034741:RNA, Small Interfering; D044767:Ubiquitin-Protein Ligases / Q000235:genetics / Q000378:metabolism*; D000097795:ATPase Inhibitory Protein; D000099202:PTEN-Induced Putative Kinase
chemical_list: D011494:Protein Kinases; D011506:Proteins; D034741:RNA, Small Interfering; D044767:Ubiquitin-Protein Ligases; D000097795:ATPase Inhibitory Protein; D000099202:PTEN-Induced Putative Kinase; C111567:parkin protein
keywords: ATPIF1; F1-Fo ATP synthase; IF1; Mitochondria; Mitophagy; PARK2; PINK1; RNA interference screen