DJ-1 is indispensable for the S-nitrosylation of Parkin, which maintains function of mitochondria

Abstract

The DJ-1 gene, a causative gene for familial Parkinson's disease (PD), has been reported to have various functions, including transcriptional regulation, antioxidant response, and chaperone and protease functions; however, the molecular mechanism associated with the pathogenesis of PD remains elusive. To further explore the molecular function of DJ-1 in the pathogenesis of PD, we compared protein expression profiles in brain tissues from wild-type and DJ-1-deficient mice. Two-dimensional difference gel electrophoresis analysis and subsequent analysis using data mining methods revealed alterations in the expression of molecules associated with energy production. We demonstrated that DJ-1 deletion inhibited S-nitrosylation of endogenous Parkin as well as overexpressed Parkin in neuroblastoma cells and mouse brain tissues. Thus, we used genome editing to generate neuroblastoma cells with DJ-1 deletion or S-nitrosylated cysteine mutation in Parkin and demonstrated that these cells exhibited similar phenotypes characterized by enhancement of cell death under mitochondrial depolarization and dysfunction of mitochondria. Our data indicate that DJ-1 is required for the S-nitrosylation of Parkin, which positively affects mitochondrial function, and suggest that the denitrosylation of Parkin via DJ-1 inactivation might contribute to PD pathogenesis and act as a therapeutic target.The protein deglycase DJ-1 is encoded by the PARK7 gene in humans 1,2 . The DJ-1 gene was originally identified as an oncogene that enhances the Ras/MAPK pathway and transforms fibroblastic cells 1 and was later identified as a causative gene for autosomal recessive juvenile parkinsonism (ARJP) 1,2 . DJ-1 has been reported to play important roles in various cellular functions, including antioxidant response mediation and mitochondrial regulation 1 . The Cys106 residue of DJ-1 is highly susceptible to oxidative stress and is oxidized, and its mutation results in loss of DJ-1 activity 3 . DJ-1 has been reported to exert neuroprotective effects at least in part via antioxidant defense; however, the mechanisms by which DJ-1 protects neurons from oxidative stress and mutation of the DJ-1 gene contributes to the pathogenesis of PD remain to be elucidated.Studies in animal and cellular models have verified that DJ-1, Parkin and pten-induced kinase 1 (PINK1) are linked. In Drosophila, deletion of parkin and pink1, which are also causative genes for ARJP, causes morphological transformation and mitochondrial dysfunction in energy-demanding tissues including muscles and brain 4 . Upregulation of parkin rescues the effects of pink1 mutation, thus indicating that these two genes are in the same genetic pathway and that parkin is downstream of pink1. Flies with dj-1 deletion exhibit similar biological phenotypes as flies with either pink1 or parkin deletion, and upregulation of fly dj-1 or human DJ-1 rescues pink1 but not parkin deficiency 5 . In primary neurons and transformed cells, overexpression of either Parkin or PINK1 rescues the mitochondrial fragmenta...

Autophagy and Redox Homeostasis in Parkinson’s: A Crucial Balancing Act

“…Crucially, through NF- κ B regulation, DJ-1 controls the expression of mitochondrial uncoupling proteins, UCP4 and UCP5, that decrease mitochondrial membrane potential, thereby suppressing ROS production; meanwhile, DJ-1 binds to Complex I via NDUFA4 to maintain its activity, and is thus crucial for mDAN survival [ 472 ]. As previously described, DJ-1 is also involved in the nitrosylation of Parkin1 [ 311 ], and DJ-1 itself can be oxidised. This is crucial for neuroprotection, and indeed, levels of oxidised DJ-1 are reduced in Parkinson's patients [ 307 , 473 ], and low levels of DJ-1 increase vulnerability to oxidative stress [ 474 ].…”

“…Conversely, in the presence of high levels of oxidative stress, DJ-1 is oxidised to the sulfonic form; this isoform is inactive and predisposed to aggregate formation, and indeed, this overoxidised isoform has been detected in brains from Parkinson's patients [ 309 , 310 ]. In addition, Ozawa et al highlighted the crucial role of DJ-1 in the nitrosylation of Parkin, and suggested that DJ-1 inactivation reduces mitophagy, leading to mitochondrial dysfunction and Parkinson's pathogenesis [ 311 ].…”

DJ-1: A promising therapeutic candidate for ischemia-reperfusion injury

“…Indeed, DJ-1 has been described to interact with complex I [ 26 , 27 ] and, more recently, with the β subunit of F 1 F O ATP synthase [ 28 ], modulating their functionality. Moreover, the protein has been found to take part in the process of mitochondrial quality control, playing a role in the Parkin/PINK1-mediated mitophagy [ [29] , [30] , [31] , [32] ], and in the regulation of the mitochondrial dynamics [ [33] , [34] , [35] , [36] ]. Noteworthy, the protein has also been suggested to modulate endoplasmic reticulum-mitochondria tethering, playing a role in the modulation of calcium transients [ 37 , 38 ].…”

The Role of DJ-1 in Cellular Metabolism and Pathophysiological Implications for Parkinson’s Disease

“…Ozawa and colleagues performed a 2D gel electrophoresis-based proteomic analysis of brain tissue from DJ-1 deficient mice and found a significant change in protein expression of pathways related to energy production including glycolysis, creatine pathway, mitochondrial TCA cycle, and ROS signaling pathway [94]. According to their analysis, spots of proteins such as PDH were decreased in DJ-1 KO compared to WT mice (Figure 1).…”