Parkin protects human dopaminergic neuroblastoma cells against dopamine-induced apoptosis

Jiang, Houbo; Ren, Yong; Zhao, Jinghui; Feng, Jian

doi:10.1093/hmg/ddh180

Cited by 228 publications

(183 citation statements)

References 36 publications

Supporting

Mentioning

160

Contrasting

Order By: Relevance

“…Parkin expression protects against caspase activation and cell death induced by stressors such as staurosporine, C2 ceramide, rotenone, and 6-OHDA (10,(12)(13)(14), and we extended these observations to include another stressor, etoposide. Neuronal dopaminergic MES and MES cells stably overexpressing human parkin (MES-Parkin) were treated with vehicle (DMSO) or etoposide (100 μM), and caspase 3/7 activity was measured at 18 h. Parental MES cells demonstrated a 100% increase in caspase 3/7 activity that was prevented by the stable over-expression of parkin (Fig.…”

Section: Resultsmentioning

confidence: 82%

The ubiquitin E3 ligase parkin regulates the proapoptotic function of Bax

Johnson

Berger²,

Cortese³

et al. 2012

Proc. Natl. Acad. Sci. U.S.A.

156

144

View full text Add to dashboard Cite

Autosomal recessive loss-of-function mutations within the PARK2 gene functionally inactivate the E3 ubiquitin ligase parkin, resulting in neurodegeneration of catecholaminergic neurons and a familial form of Parkinson disease. Current evidence suggests both a mitochondrial function for parkin and a neuroprotective role, which may in fact be interrelated. The antiapoptotic effects of parkin have been widely reported, and may involve fundamental changes in the threshold for apoptotic cytochrome c release, but the substrate(s) involved in parkin dependent protection had not been identified. Here, we demonstrate the parkin-dependent ubiquitination of endogenous Bax comparing primary cultured neurons from WT and parkin KO mice and using multiple parkin-overexpressing cell culture systems. The direct ubiquitination of purified Bax was also observed in vitro following incubation with recombinant parkin. We found that parkin prevented basal and apoptotic stressinduced translocation of Bax to the mitochondria. Moreover, an engineered ubiquitination-resistant form of Bax retained its apoptotic function, but Bax KO cells complemented with lysine-mutant Bax did not manifest the antiapoptotic effects of parkin that were observed in cells expressing WT Bax. These data suggest that Bax is the primary substrate responsible for the antiapoptotic effects of parkin, and provide mechanistic insight into at least a subset of the mitochondrial effects of parkin.Parkinson's disease | apoptosis | neuroprotection | mitophagy P arkinson disease (PD) is a neurodegenerative disorder that affects 1-3% of the population over the age of 65 years (1). The symptoms include tremor, rigidity, bradykinesia, and postural instability. These physical characteristics are caused by the progressive degeneration of dopaminergic neurons of the substantia nigra pars compacta and, to a lesser extent, the catecholaminergic neurons of the locus coeruleus. Although most cases of PD are sporadic in nature, a small number of genes are responsible for the rare familial forms of PD (2). Loss-of-function mutations within the PARK2 locus, which encodes the protein parkin, are the most common cause of autosomal recessive PD (3).Parkin is a 465-amino acid protein that is expressed in multiple tissues and functions as an E3 ubiquitin ligase (4). Ubiquitination of substrates is a tightly regulated process, requiring the combined activity of three enzymes: an E1 ubiquitin-activating enzyme, an E2 ubiquitin conjugating enzyme, and an E3 ubiquitin ligase (5). E3 ubiquitin ligases are responsible for substrate recognition, and as such contribute the specificity of a ubiquitin reaction. Defects in parkin-mediated ubiquitination may result in the failure to target specific substrates for degradation, leading to accumulation of potentially toxic proteins and consequent cell death (6). Parkin is widely neuroprotective (7); however, many of the putative parkin substrates reported to date are not thought to directly mediate toxicity in such a simple fashion [reviewed elsewhere (8...

show abstract

Section: Resultsmentioning

confidence: 82%

The ubiquitin E3 ligase parkin regulates the proapoptotic function of Bax

Johnson

Berger²,

Cortese³

et al. 2012

Proc. Natl. Acad. Sci. U.S.A.

156

144

View full text Add to dashboard Cite

show abstract

“…Our finding that altered GstS1 activity inf luences the parkin mutant phenotypes raises the possibility that parkin may also offer protection from the effects of oxidative stress. Further support for this hypothesis comes from the findings that oxidative damage in f lies, mice, and cell lines and sensitivity to oxidative stress agents in f lies and cell lines correlates inversely with parkin activity (19,(32)(33)(34). Our recent results from transcriptional profiling of parkin mutants and a genetic screen for parkin modifiers also demonstrate that oxidative stress response elements are up-regulated and that mutations in oxidative stress response components enhance the parkin mutant phenotypes (15).…”

Section: Discussionmentioning

confidence: 89%

Increased glutathione S -transferase activity rescues dopaminergic neuron loss in a Drosophila model of Parkinson's disease

Whitworth

Theodore²,

Greene

et al. 2005

Proc. Natl. Acad. Sci. U.S.A.

388

373

View full text Add to dashboard Cite

Loss-of-function mutations of the parkin gene are a major cause of early-onset parkinsonism. To explore the mechanism by which loss of parkin function results in neurodegeneration, we are using a genetic approach in Drosophila. Here, we show that Drosophila parkin mutants display degeneration of a subset of dopaminergic (DA) neurons in the brain. The neurodegenerative phenotype of parkin mutants is enhanced by loss-of-function mutations of the glutathione S-transferase S1 (GstS1) gene, which were identified in an unbiased genetic screen for genes that modify parkin phenotypes. Furthermore, overexpression of GstS1 in DA neurons suppresses neurodegeneration in parkin mutants. Given the previous evidence for altered glutathione metabolism and oxidative stress in sporadic Parkinson's disease (PD), these data suggest that the mechanism of DA neuron loss in Drosophila parkin mutants is similar to the mechanisms underlying sporadic PD. Moreover, these findings identify a potential therapeutic approach in treating PD.genetic modifier ͉ neurodegeneration ͉ parkin P arkinson's disease (PD) is a common neurodegenerative disorder characterized by the loss of dopaminergic (DA) neurons in the substantia nigra and the accumulation of proteinaceous intraneuronal inclusions known as Lewy bodies. The mechanisms responsible for neurodegeneration in PD are largely unknown, although previous work suggests that mitochondrial complex I dysfunction, oxidative stress, and aberrant proteolytic degradation may contribute to pathogenesis (1). The recent identification of genes responsible for rare inherited forms of parkinsonism presents an opportunity to establish neurodegenerative mechanisms that may be relevant to sporadic forms of PD.Loss-of-function mutations of parkin are a common cause of autosomal recessive juvenile parkinsonism (ARJP), and parkin dysfunction may also contribute to late-onset sporadic PD (2-6). Patients with parkin mutations display many of the typical features of idiopathic PD, including locomotor dysfunction, reduced mitochondrial complex I activity, and degeneration of DA neurons in the substantia nigra. However, most ARJP cases have a significantly earlier age of onset and lack Lewy body pathology. Parkin has been shown to possess ubiquitin-protein ligase activity (7-9), which acts to confer substrate target specificity in the ubiquitin͞proteasome protein degradation pathway. This finding has led to the model that toxic accumulation of parkin substrates may be responsible for DA neuron death. A number of putative substrates of parkin have been identified (10). Several of these parkin substrates, including the Lewy body component ␣-synuclein (11) and the putative G protein-coupled receptor Pael-R (12), have received considerable attention, in part because they implicate specific cellular pathways in parkinmediated pathogenesis. However, the involvement of many of the identified parkin substrates in the etiology of ARJP remains controversial.To identify pathways relevant to parkin pathogenesis, we are using a gen...

show abstract

“…Thus, abnormal ERK1/2 activation plays a critical role in PD pathogenesis. Parkin is documented to mediate neuroprotection against 6-OHDA-induced neurotoxicity [7,21,41,42] . As parkin is an E3 ubiquitin ligase, its neuroprotective effect may be executed via its substrates.…”

Section: Discussionmentioning

confidence: 99%

“…PARK2 mutations lead to the loss of its function and therefore to an accumulation of its substrates in the brain; such accumulation is related to the occurrence of PD [6] . It has been reported that parkin, but not its pathogenic mutants, can protect DA neurons against neurotoxicity [7] .…”

Section: Introductionmentioning

confidence: 99%

Parkin represses 6-hydroxydopamine-induced apoptosis via stabilizing scaffold protein p62 in PC12 cells

Hou

Ren

et al. 2015

Acta Pharmacol Sin

View full text Add to dashboard Cite

Aim: Parkin has been shown to exert protective effects against 6-hydroxydopamine (6-OHDA)-induced neurotoxicity in different models of Parkinson disease. In the present study we investigated the molecular mechanisms underlying the neuroprotective action of parkin in vitro. Methods: HEK293, HeLa and PC12 cells were transfected with parkin, parkin mutants, p62 or si-p62. Protein expression and ubiquitination were assessed using immunoblot analysis. Immunoprecipitation assay was performed to identify the interaction between parkin and scaffold protein p62. PC12 and SH-SY5Y cells were treated with 6-OHDA (200 μmol/L), and cell apoptosis was detected using PI and Hoechst staining. Results: In HEK293 cells co-transfected with parkin and p62, parkin was co-immunoprecipitated with p62, and parkin overexpression increased p62 protein levels. In parkin-deficient HeLa cells, transfection with wild-type pakin, but not with ligase activity-deficient pakin mutants, significantly increased p62 levels, suggesting that parkin stabilized p62 through its E3 ligase activity. Transfection with parkin or p62 significantly repressed ERK1/2 phosphorylation in HeLa cells, but transfection with parkin did not repress ERK1/2 phosphorylation in p62-knockdown HeLa cells, suggesting that p62 was involved in parkin-induced inhibition on ERK1/2 phosphorylation. Overexpression of parkin or p62 significantly repressed 6-OHDA-induced ERK1/2 phosphorylation in PC12 cells, and parkin overexpression inhibited 6-OHDA-induced apoptosis in PC12 and SH-SY5Y cells. Conclusion: Parkin protects PC12 cells against 6-OHDA-induced apoptosis via ubiquitinating and stabilizing scaffold protein p62, and repressing ERK1/2 activation.

show abstract

Parkin protects human dopaminergic neuroblastoma cells against dopamine-induced apoptosis

Cited by 228 publications

References 36 publications

The ubiquitin E3 ligase parkin regulates the proapoptotic function of Bax

The ubiquitin E3 ligase parkin regulates the proapoptotic function of Bax

Increased glutathione S -transferase activity rescues dopaminergic neuron loss in a Drosophila model of Parkinson's disease

Parkin represses 6-hydroxydopamine-induced apoptosis via stabilizing scaffold protein p62 in PC12 cells

Contact Info

Product

Resources

About