Stress-induced alterations in parkin solubility promote parkin aggregation and compromise parkin's protective function

Wang, Cheng; Ko, Han Seok; Thomas, Bobby; Tsang, Fai; Chew, Katherine C. M.; Tay, Shiam Peng; Ho, Michelle W L; Lim, Tit Meng; Soong, Tuck Wah; Pletniková, Olga; Troncoso, Juan C.; Dawson, Valina L.; Dawson, Ted M.; Lim, Kah Leong

doi:10.1093/hmg/ddi413

Cited by 196 publications

(166 citation statements)

References 42 publications

Supporting

Mentioning

147

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 findings provide further support to the idea that parkin is inactivated in sporadic PD. Previous studies suggest that parkin is inactivated in sporadic PD through S-nitrosylation (15,26), oxidative stress (27), and dopamine conjugation (18). Here we describe another posttranslational modification of parkin that is present in sporadic PD.…”

Section: Discussionmentioning

confidence: 99%

Phosphorylation by the c-Abl protein tyrosine kinase inhibits parkin's ubiquitination and protective function

Ko¹,

Lee²,

Shin³

et al. 2010

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

Self Cite

246

291

View full text Add to dashboard Cite

Mutations in PARK2/Parkin, which encodes a ubiquitin E3 ligase, cause autosomal recessive Parkinson disease (PD). Here we show that the nonreceptor tyrosine kinase c-Abl phosphorylates tyrosine 143 of parkin, inhibiting parkin's ubiquitin E3 ligase activity and protective function. c-Abl is activated by dopaminergic stress and by dopaminergic neurotoxins, 1-methyl-4-phenylpyridinium (MPP + ) in vitro and in vivo by 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP), leading to parkin inactivation, accumulation of the parkin substrates aminoacyl-tRNA synthetase-interacting multifunctional protein type 2 (AIMP2) (p38/JTV-1) and fuse-binding protein 1 (FBP1), and cell death. STI-571, a c-Abl-family kinase inhibitor, prevents the phosphorylation of parkin, maintaining parkin in a catalytically active and protective state. STI-571's protective effects require parkin, as shRNA knockdown of parkin prevents STI-571 protection. Conditional knockout of c-Abl in the nervous system also prevents the phosphorylation of parkin, the accumulation of its substrates, and subsequent neurotoxicity in response to MPTP intoxication. In human postmortem PD brain, c-Abl is active, parkin is tyrosine-phosphorylated, and AIMP2 and FBP1 accumulate in the substantia nigra and striatum. Thus, tyrosine phosphorylation of parkin by c-Abl is a major posttranslational modification that inhibits parkin function, possibly contributing to pathogenesis of sporadic PD. Moreover, inhibition of c-Abl may be a neuroprotective approach in the treatment of PD.is a common neurodegenerative disorder characterized by the loss of dopamine (DA) neurons and protein accumulation in intracellular inclusions designated as Lewy bodies and Lewy neurites (1). Although the majority of PD is sporadic in nature, rare familial mutations are providing insight into this chronic, progressive neurodegenerative disease. Mutations in α-synuclein and LRRK2 cause autosomal-dominant PD, whereas mutations in DJ-1, PINK1, and parkin result in autosomal-recessive PD (2). Parkin mutations are the most common cause of autosomal-recessive PD and, for the most part, PD due to parkin mutations is indistinguishable from sporadic PD (3). Parkin is a ubiquitin E3 ligase, and familial mutations are thought to impair the E3 ligase activity of parkin (4, 5).Parkin ubiquitinates proteins via monoubiquitination or polyubiquitination using either lysine 48 (K48) or lysine 63 (K63). Monoubiquitination by parkin is thought to regulate receptor trafficking (6). Polyubiquitination by parkin via K48 is thought to mediate proteasomal degradation (7,8), whereas polyubiquitination by K63 may be involved in inclusion formation (9). Parkin's differential ubiquitination properties are likely to be regulated by different ubiquitin-conjugating E2s and other associated proteins or regulatory processes (3). A number of putative parkin substrates have been identified (for a review, see ref.3). Aminoacyl-tRNA synthetaseinteracting multifunctional protein type 2 (AIMP2) (p38/JTV-1) and fuse-binding protein 1 (FB...

show abstract

“…32). Misfolding of Parkin induced by pathogenic mutations or cellular stress has been established as a major mechanism of Parkin inactivation, accentuating a possible pathological role of Parkin even in sporadic PD (11)(12)(13)(14)(15)(16)(17)(18)(19)(20)(21)(33)(34)(35). It is obvious that the high cysteine content found in the RBR domain predisposes Parkin to oxidative stress-induced inactivation and misfolding, a phenomenon that has recently been shown experimentally (36).…”

Section: Discussionmentioning

confidence: 99%

Aberrant Folding of Pathogenic Parkin Mutants

Schlehe

Lutz

Pilsl

et al. 2008

Journal of Biological Chemistry

View full text Add to dashboard Cite

Parkinson disease (PD)2 is the second most common neurodegenerative disease after Alzheimer disease. Although most PD cases occur sporadically, familial variants share important features with sporadic PD, most notably the demise of dopaminergic neurons in the substantia nigra pars compacta. Consequently, insight into the function of PD-associated genes might promote our understanding of pathogenic mechanisms not only in familial, but also in sporadic PD. Five genes have unambiguously been linked to PD over the past decade, the genes encoding ␣-synuclein and LRRK2 for autosomal dominant PD, and those encoding Parkin, PINK1, and DJ-1 for autosomal recessive PD (reviewed in Refs. 1-3). So far, over a hundred different pathogenic mutations in the parkin gene have been identified, which account for the majority of autosomal recessive PD cases. Parkin is a member of the RBR (ring between ring fingers) protein family, characterized by the presence of two RING domains (really interesting new gene), which flank a cysteine-rich in-between RINGs (IBR) domain. Similarly to other RBR proteins, Parkin has an E3 ubiquitin ligase activity, mediating the attachment of ubiquitin to substrate proteins (4 -6). Parkin can obviously mediate different modes of ubiquitylation, including monoubiquitylation, multiple monoubiquitylation, and polyubiquitylation both via lysine 48 and lysine 63, depending on the experimental conditions and the putative Parkin substrate analyzed (reviewed in Refs. 7 and 8). Importantly, the neuroprotective activity of Parkin seems to be associated with its ability to promote degradation-independent ubiquitylation (9, 10).Different lines of evidence indicate that pathogenic parkin mutations result in a loss of Parkin function. Our initial studies revealed that misfolding and aggregation is characteristic for C-terminal deletion mutants of Parkin based on the following biochemical features specific for mutant Parkin: 1) insolubility in non-ionic and ionic detergents; 2) sedimentation in a sucrose step gradient; 3) resistance to a limited proteolytic digestion; 4) loss of membrane association; and 5) formation of scattered aggregates in cells determined by immunocytochemistry (11,12). Alterations in the detergent solubility of Parkin and formation of Parkin aggregates/inclusion bodies have also been reported for various Parkin missense mutants (13-18). We also observed that even wild-type Parkin is prone to misfolding under severe oxidative stress (12). Remarkably, insoluble, catechol-modified Parkin could be detected in the substantia nigra of patients suffering from sporadic PD, suggesting a more general role of Parkin in the pathogenesis of PD (19). In support of this concept, the E3 ligase activity of Parkin has been shown to be impaired by nitrosative stress, and there is indeed evidence for the presence of S-nitrosylated Parkin in the brains of PD patients (20,21).Based on our finding that the deletion of C-terminal amino acids results in misfolding and aggregation of Parkin, we performed a comparative an...

show abstract

Stress-induced alterations in parkin solubility promote parkin aggregation and compromise parkin's protective function

Cited by 196 publications

References 42 publications

The ubiquitin E3 ligase parkin regulates the proapoptotic function of Bax

The ubiquitin E3 ligase parkin regulates the proapoptotic function of Bax

Phosphorylation by the c-Abl protein tyrosine kinase inhibits parkin's ubiquitination and protective function

Aberrant Folding of Pathogenic Parkin Mutants

Contact Info

Product

Resources

About