title: High-content genome-wide RNAi screens identify regulators of parkin upstream of mitophagy.
abstract: An increasing body of evidence points to mitochondrial dysfunction as a contributor to the molecular pathogenesis of neurodegenerative diseases such as Parkinson's disease. Recent studies of the Parkinson's disease associated genes PINK1 (ref. 2) and parkin (PARK2, ref. 3) indicate that they may act in a quality control pathway preventing the accumulation of dysfunctional mitochondria. Here we elucidate regulators that have an impact on parkin translocation to damaged mitochondria with genome-wide small interfering RNA (siRNA) screens coupled to high-content microscopy. Screening yielded gene candidates involved in diverse cellular processes that were subsequently validated in low-throughput assays. This led to characterization of TOMM7 as essential for stabilizing PINK1 on the outer mitochondrial membrane following mitochondrial damage. We also discovered that HSPA1L (HSP70 family member) and BAG4 have mutually opposing roles in the regulation of parkin translocation. The screens revealed that SIAH3, found to localize to mitochondria, inhibits PINK1 accumulation after mitochondrial insult, reducing parkin translocation. Overall, our screens provide a rich resource to understand mitochondrial quality control.
authors.affiliation: 1] Surgical Neurology Branch, National Institute of Neurological Disorders and Stroke, National Institutes of Health, Bethesda, Maryland 20892, USA [2] Division of Preclinical Innovation, National Center for Advancing Translational Sciences, National Institutes of Health, Rockville, Maryland 20850, USA [3].
authors.forename: Samuel A
authors.initials: SA
authors.lastname: Hasson
authors.affiliation: 1] Surgical Neurology Branch, National Institute of Neurological Disorders and Stroke, National Institutes of Health, Bethesda, Maryland 20892, USA [2].
authors.forename: Lesley A
authors.initials: LA
authors.lastname: Kane
authors.affiliation: Surgical Neurology Branch, National Institute of Neurological Disorders and Stroke, National Institutes of Health, Bethesda, Maryland 20892, USA.
authors.forename: Koji
authors.initials: K
authors.lastname: Yamano
authors.affiliation: Surgical Neurology Branch, National Institute of Neurological Disorders and Stroke, National Institutes of Health, Bethesda, Maryland 20892, USA.
authors.forename: Chiu-Hui
authors.initials: CH
authors.lastname: Huang
authors.affiliation: Surgical Neurology Branch, National Institute of Neurological Disorders and Stroke, National Institutes of Health, Bethesda, Maryland 20892, USA.
authors.forename: Danielle A
authors.initials: DA
authors.lastname: Sliter
authors.affiliation: Division of Preclinical Innovation, National Center for Advancing Translational Sciences, National Institutes of Health, Rockville, Maryland 20850, USA.
authors.forename: Eugen
authors.initials: E
authors.lastname: Buehler
authors.affiliation: Surgical Neurology Branch, National Institute of Neurological Disorders and Stroke, National Institutes of Health, Bethesda, Maryland 20892, USA.
authors.forename: Chunxin
authors.initials: C
authors.lastname: Wang
authors.affiliation: NIH Center for Regenerative Medicine, Bethesda, Maryland 20892, USA.
authors.forename: Sabrina M
authors.initials: SM
authors.lastname: Heman-Ackah
authors.affiliation: Surgical Neurology Branch, National Institute of Neurological Disorders and Stroke, National Institutes of Health, Bethesda, Maryland 20892, USA.
authors.forename: Tara
authors.initials: T
authors.lastname: Hessa
authors.affiliation: Division of Preclinical Innovation, National Center for Advancing Translational Sciences, National Institutes of Health, Rockville, Maryland 20850, USA.
authors.forename: Rajarshi
authors.initials: R
authors.lastname: Guha
authors.affiliation: Division of Preclinical Innovation, National Center for Advancing Translational Sciences, National Institutes of Health, Rockville, Maryland 20850, USA.
authors.forename: Scott E
authors.initials: SE
authors.lastname: Martin
authors.affiliation: Surgical Neurology Branch, National Institute of Neurological Disorders and Stroke, National Institutes of Health, Bethesda, Maryland 20892, USA.
authors.forename: Richard J
authors.initials: RJ
authors.lastname: Youle
mesh_terms: D048868:Adaptor Proteins, Signal Transducing / Q000378:metabolism; D015894:Genome, Human / Q000235:genetics*; D045325:HCT116 Cells; D057809:HEK293 Cells; D018840:HSP70 Heat-Shock Proteins / Q000378:metabolism; D006367:HeLa Cells; D006801:Humans; D008565:Membrane Proteins / Q000378:metabolism; D008928:Mitochondria / Q000378:metabolism / Q000473:pathology; D051336:Mitochondrial Membranes / Q000378:metabolism; D000090622:Mitochondrial Precursor Protein Import Complex Proteins; D024101:Mitochondrial Proteins / Q000378:metabolism; D063306:Mitophagy; D005810:Multigene Family / Q000235:genetics; D010300:Parkinson Disease / Q000378:metabolism / Q000473:pathology; D011494:Protein Kinases / Q000378:metabolism; D021381:Protein Transport; D034622:RNA Interference; D034741:RNA, Small Interfering / Q000032:analysis / Q000235:genetics; D015203:Reproducibility of Results; D044767:Ubiquitin-Protein Ligases / Q000378:metabolism*; D000099202:PTEN-Induced Putative Kinase
chemical_list: D048868:Adaptor Proteins, Signal Transducing; D018840:HSP70 Heat-Shock Proteins; D008565:Membrane Proteins; D000090622:Mitochondrial Precursor Protein Import Complex Proteins; D024101:Mitochondrial Proteins; D011494:Protein Kinases; D034741:RNA, Small Interfering; D044767:Ubiquitin-Protein Ligases; D000099202:PTEN-Induced Putative Kinase; C116906:BAG4 protein, human; C543454:HSPA1L protein, human; C405562:TOMM7 protein, human; C111567:parkin protein