1. High-content genome-wide RNAi screens identify regulators of parkin upstream of mitophagy
- Author
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Hasson, Samuel A., Kane, Lesley A., Yamano, Koji, Huang, Chiu-Hui, Sliter, Danielle A., Buehler, Eugen, Wang, Chunxin, Heman-Ackah, Sabrina M., Hessa, Tara, Guha, Rajarshi, Martin, Scott E., and Youle, Richard J.
- Subjects
Genomics -- Research ,Autophagy (Cytology) -- Physiological aspects -- Genetic aspects ,Molecular biology -- Research ,Mitochondria -- Genetic aspects -- Health aspects -- Physiological aspects ,Parkinson's disease -- Physiological aspects -- Genetic aspects ,RNA -- Health aspects ,Genetic regulation -- Health aspects ,Environmental issues ,Science and technology ,Zoology and wildlife conservation - 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 (1). Recent studies of the Parkinson's disease associated genes PINK1 (ref. 2) and parkin (PARK, ref. 3) indicate that they may act in a quality control pathway preventing the accumulation of dysfunctional mitochondria (4-8). 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., Following the loss of mitochondrial membrane potential, PINK1 and parkin coordinate a ubiquitination (9), proteasomal activation (10) and autophagic (mitophagy) (5) response that may attenuate cell death (11). As mitophagy [...]
- Published
- 2013