Effect of endogenous mutant and wild-type PINK1 on Parkin in fibroblasts from Parkinson disease patients.
Mutations in the PTEN-induced putative kinase 1 (PINK1), a mitochondrial serine-threonine kinase, and Parkin, an E3 ubiquitin ligase, are associated with autosomal-recessive forms of Parkinson disease (PD). Both are involved in the maintenance of mitochondrial integrity and protection from multiple stressors. Recently, Parkin was demonstrated to be recruited to impaired mitochondria in a PINK1-dependent manner, where it triggers mitophagy. Using primary human dermal fibroblasts originating from PD patients with various PINK1 mutations, we showed at the endogenous level that (i) PINK1 regulates the stress-induced decrease of endogenous Parkin; (ii) mitochondrially localized PINK1 mediates the stress-induced mitochondrial translocation of Parkin; (iii) endogenous PINK1 is stabilized on depolarized mitochondria; and (iv) mitochondrial accumulation of full-length PINK1 is sufficient but not necessary for the stress-induced loss of Parkin signal and its mitochondrial translocation. Furthermore, we showed that different stressors, depolarizing or non-depolarizing, led to the same effect on detectable Parkin levels and its mitochondrial targeting. Although this effect on Parkin was independent of the mitochondrial membrane potential, we demonstrate a differential effect of depolarizing versus non-depolarizing stressors on endogenous levels of PINK1. Our study shows the necessity to introduce an environmental factor, i.e. stress, to visualize the differences in the interaction of PINK1 and Parkin in mutants versus controls. Establishing human fibroblasts as a suitable model for studying this interaction, we extend data from animal and other cellular models and provide experimental evidence for the generally held notion of PD as a condition with a combined genetic and environmental etiology.
Rakovic
Aleksandar
A
Section of Clinical and Molecular Neurogenetics, Department of Neurology, University of Lübeck, 23562 Lübeck, Germany.
Grünewald
Anne
A
Seibler
Philip
P
Ramirez
Alfredo
A
Kock
Norman
N
Orolicki
Slobodanka
S
Lohmann
Katja
K
Klein
Christine
C
D015153:Blotting, Western; D002460:Cell Line; D002478:Cells, Cultured; D015853:Cysteine Proteinase Inhibitors / Q000494:pharmacology; D003600:Cytosol / Q000378:metabolism; D005347:Fibroblasts / Q000187:drug effects / Q000378:metabolism*; D006801:Humans; D006861:Hydrogen Peroxide / Q000494:pharmacology; D007476:Ionophores / Q000494:pharmacology; D007976:Leupeptins / Q000494:pharmacology; D008928:Mitochondria / Q000378:metabolism; D009154:Mutation; D009842:Oligopeptides / Q000494:pharmacology; D016877:Oxidants / Q000494:pharmacology; D010300:Parkinson Disease / Q000235:genetics / Q000378:metabolism / Q000473:pathology; D011494:Protein Kinases / Q000235:genetics / Q000378:metabolism*; D021381:Protein Transport / Q000187:drug effects; D034622:RNA Interference; D014162:Transfection; D044767:Ubiquitin-Protein Ligases / Q000235:genetics / Q000378:metabolism*; D014634:Valinomycin / Q000494:pharmacology; D000099202:PTEN-Induced Putative Kinase
D015853:Cysteine Proteinase Inhibitors; D006861:Hydrogen Peroxide; D007476:Ionophores; D007976:Leupeptins; D009842:Oligopeptides; D016877:Oxidants; D011494:Protein Kinases; D044767:Ubiquitin-Protein Ligases; D014634:Valinomycin; D000099202:PTEN-Induced Putative Kinase; C111567:parkin protein; C072553:benzyloxycarbonylleucyl-leucyl-leucine aldehyde; C078846:epoxomicin