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

Nature. 2013 Dec 12;504(7479):291-5. doi: 10.1038/nature12748. Epub 2013 Nov 24.

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.

Publication types

  • Research Support, N.I.H., Intramural
  • Research Support, Non-U.S. Gov't

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
  • Mitochondria / pathology
  • Mitochondrial Membranes / metabolism
  • Mitochondrial Proteins / metabolism
  • Mitophagy*
  • Multigene Family / genetics
  • Parkinson Disease / metabolism
  • Parkinson Disease / pathology
  • Protein Kinases / metabolism
  • Protein Transport
  • RNA Interference*
  • RNA, Small Interfering / analysis
  • RNA, Small Interfering / genetics
  • Reproducibility of Results
  • Ubiquitin-Protein Ligases / metabolism*

Substances

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