The serine protease HtrA2 cleaves UCH-L1 and inhibits its hydrolase activity: Implication in the UCH-L1-mediated cell death

Park, Dae-Wook; Nam, Min-Kyung; Rhim, Hyangshuk

doi:10.1016/j.bbrc.2011.09.148

Cited by 9 publications

(10 citation statements)

References 30 publications

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“…Since a previous study had shown that UCH-L1 is cleaved by HtrA2/Omi during staurosporine-induced apoptosis [38], we investigated whether UCH-L1 also served as a substrate and thus potential downstream effector of HtrA2/Omi in TNF-induced necroptosis. Initially supporting this assumption, Western blots revealed a decrease of the 25-kDa band representing full-length UCH-L in lysates from wild-type (WT) MEF after induction of necroptosis by TNF/zVAD/CHX (but not in untreated or zVAD/CHX-treated controls, Figure 4A).…”

Section: Resultsmentioning

confidence: 99%

“…Park and colleagues have reported that HtrA2/Omi cleaves UCH-L1 during staurosporine-induced apoptosis, generating a 10-kDa cleavage fragment (although this was shown only in vitro and upon overexpression, but not for the endogenous proteins) [38]. We therefore included positive controls for cleavage of endogenous UCH-L1 by endogenous HtrA2/Omi by treating WT MEF with staurosporine, and additionally compared them to staurosporine-treated HtrA2/Omi-deficient MEF.…”

Section: Resultsmentioning

confidence: 99%

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The proteases HtrA2/Omi and UCH-L1 regulate TNF-induced necroptosis

et al. 2013

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BackgroundIn apoptosis, proteolysis by caspases is the primary mechanism for both initiation and execution of programmed cell death (PCD). In contrast, the impact of proteolysis on the regulation and execution of caspase-independent forms of PCD (programmed necrosis, necroptosis) is only marginally understood. Likewise, the identity of the involved proteases has remained largely obscure. Here, we have investigated the impact of proteases in TNF-induced necroptosis.ResultsThe serine protease inhibitor TPKC protected from TNF-induced necroptosis in multiple murine and human cells systems whereas inhibitors of metalloproteinases or calpain/cysteine and cathepsin proteases had no effect. A screen for proteins labeled by a fluorescent TPCK derivative in necroptotic cells identified HtrA2/Omi (a serine protease previously implicated in PCD) as a promising candidate. Demonstrating its functional impact, pharmacological inhibition or genetic deletion of HtrA2/Omi protected from TNF-induced necroptosis. Unlike in apoptosis, HtrA2/Omi did not cleave another protease, ubiquitin C-terminal hydrolase (UCH-L1) during TNF-induced necroptosis, but rather induced monoubiquitination indicative for UCH-L1 activation. Correspondingly, pharmacologic or RNA interference-mediated inhibition of UCH-L1 protected from TNF-induced necroptosis. We found that UCH-L1 is a mediator of caspase-independent, non-apoptotic cell death also in diseased kidney podocytes by measuring cleavage of the protein PARP-1, caspase activity, cell death and cell morphology. Indicating a role of TNF in this process, podocytes with stably downregulated UCH-L1 proved resistant to TNF-induced necroptosis.ConclusionsThe proteases HtrA2/Omi and UCH-L1 represent two key components of TNF-induced necroptosis, validating the relevance of proteolysis not only for apoptosis, but also for caspase-independent PCD. Since UCH-L1 clearly contributes to the non-apoptotic death of podocytes, interference with the necroptotic properties of HtrA2/Omi and UCH-L1 may prove beneficial for the treatment of patients, e.g. in kidney failure.

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Section: Resultsmentioning

confidence: 99%

Section: Resultsmentioning

confidence: 99%

The proteases HtrA2/Omi and UCH-L1 regulate TNF-induced necroptosis

et al. 2013

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“…UCHL1-mediated cell death can be attenuated by mitochondrial protein HTRA2 [94], ATP13A2 regulates mitochondrial bioenergetics through macroautophagy [95], VPS35 mediates vesicle transport between mitochondria and peroxisomes [96], and EIF4G1 is involved in stress related protection of mitochondria [97]. …”

Section: Mitochondrial Dysfunctionmentioning

confidence: 99%

Integrating Pathways of Parkinson's Disease in a Molecular Interaction Map

et al. 2013

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Parkinson's disease (PD) is a major neurodegenerative chronic disease, most likely caused by a complex interplay of genetic and environmental factors. Information on various aspects of PD pathogenesis is rapidly increasing and needs to be efficiently organized, so that the resulting data is available for exploration and analysis. Here we introduce a computationally tractable, comprehensive molecular interaction map of PD. This map integrates pathways implicated in PD pathogenesis such as synaptic and mitochondrial dysfunction, impaired protein degradation, alpha-synuclein pathobiology and neuroinflammation. We also present bioinformatics tools for the analysis, enrichment and annotation of the map, allowing the research community to open new avenues in PD research. The PD map is accessible at http://minerva.uni.lu/pd_map.Electronic supplementary materialThe online version of this article (doi:10.1007/s12035-013-8489-4) contains supplementary material, which is available to authorized users.

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“…Previous reports have indicated that Omi does locate in the nuclei [1] . To date, extensive studies have revealed the mitochondrial and cytoplasmic substrates of Omi and shown the function of Omi in these subcellular sites [4,39,40] ; however, the nuclear substrates and functions of Omi have not been fully explored. Omi regulates transcription by processing the transcription factor, p73, or the transcriptional regulator, Wilm's suppressor protein [2,3] , to regulate apoptosis.…”

Section: Discussionmentioning

confidence: 99%

Protease Omi facilitates neurite outgrowth in mouse neuroblastoma N2a cells by cleaving transcription factor E2F1

et al. 2015

Acta Pharmacol Sin

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Aim:Omi is an ATP-independent serine protease that is necessary for neuronal function and survival. The aim of this study was to investigate the role of protease Omi in regulating differentiation of mouse neuroblastoma cells and to identify the substrate of Omi involved in this process. Methods: Mouse neuroblastoma N2a cells and Omi protease-deficient mnd2 mice were used in this study. To modulate Omi and E2F1 expression, N2a cells were transfected with expression plasmids, shRNA plasmids or siRNA. Protein levels were detected using immunoblot assays. The interaction between Omi and E2F1 was studied using immunoprecipitation, GST pulldown and in vitro cleavage assays. N2a cells were treated with 20 μmol/L retinoic acid (RA) and 1% fetal bovine serum to induce neurite outgrowth, which was measured using Image J software. Results: E2F1 was significantly increased in Omi knockdown cells and in brain lysates of mnd2 mice, and was decreased in cells overexpressing wild-type Omi, but not inactive Omi S276C. In brain lysates of mnd2 mice, endogenous E2F1 was co-immunoprecipitated with endogenous Omi. In vitro cleavage assay demonstrated that Omi directly cleaved E2F1. Treatment of N2a cells with RA induced marked differentiation and neurite outgrowth accompanied by significantly increased Omi and decreased E2F1 levels, which were suppressed by pretreatment with the specific Omi inhibitor UCF-101. Knockdown of Omi in N2a cells suppressed RA-induced neurite outgrowth, which was partially restored by knockdown of E2F1. Conclusion: Protease Omi facilitates neurite outgrowth by cleaving the transcription factor E2F1 in differentiated neuroblastoma cells; E2F1 is a substrate of Omi.

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The serine protease HtrA2 cleaves UCH-L1 and inhibits its hydrolase activity: Implication in the UCH-L1-mediated cell death

Cited by 9 publications

References 30 publications

The proteases HtrA2/Omi and UCH-L1 regulate TNF-induced necroptosis

The proteases HtrA2/Omi and UCH-L1 regulate TNF-induced necroptosis

Integrating Pathways of Parkinson's Disease in a Molecular Interaction Map

Protease Omi facilitates neurite outgrowth in mouse neuroblastoma N2a cells by cleaving transcription factor E2F1

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