Home LiteratureArticle Details

PMID: 24270810 Published · ppublish English Journal Article Research Support, N.I.H., Intramural Research Support, Non-U.S. Gov't

High-content genome-wide RNAi screens identify regulators of parkin upstream of mitophagy.

Nature ·Vol. 504 ·No. 7479 ·2013-12-12 ·Pages 291-5

Hasson SA, Kane LA, Yamano K, Huang CH, Sliter DA, Buehler E, Wang C, Heman-Ackah SM, Hessa T, Guha R, Martin SE, Youle RJ

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.

MeSH Terms

Adaptor Proteins, Signal Transducing/metabolism Genome, Human/genetics HCT116 Cells HEK293 Cells HSP70 Heat-Shock Proteins/metabolism HeLa Cells Humans Membrane Proteins/metabolism Mitochondria/metabolism,pathology Mitochondrial Membranes/metabolism Mitochondrial Precursor Protein Import Complex Proteins Mitochondrial Proteins/metabolism Mitophagy Multigene Family/genetics Parkinson Disease/metabolism,pathology Protein Kinases/metabolism Protein Transport RNA Interference RNA, Small Interfering/analysis,genetics Reproducibility of Results Ubiquitin-Protein Ligases/metabolism

Chemicals

Adaptor Proteins, Signal Transducing BAG4 protein, human HSP70 Heat-Shock Proteins HSPA1L protein, human Membrane Proteins Mitochondrial Precursor Protein Import Complex Proteins Mitochondrial Proteins RNA, Small Interfering TOMM7 protein, human Ubiquitin-Protein Ligases parkin protein Protein Kinases PTEN-induced putative kinase

Authors & Affiliations

12 authors, click to expand affiliations / ORCID

Hasson Samuel A

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].

Kane Lesley A

1] Surgical Neurology Branch, National Institute of Neurological Disorders and Stroke, National Institutes of Health, Bethesda, Maryland 20892, USA [2].

Yamano Koji