ORCID Profile
0000-0001-9886-0989
Current Organisation
University of South Australia
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Publisher: Springer Science and Business Media LLC
Date: 14-12-2008
Abstract: The activation of caspases is the principal event in the execution of apoptosis. Initiator caspases are activated through an autocatalytic mechanism often involving dimerisation or oligomerisation. In Drosophila, the only initiator caspase DRONC, is tightly inhibited by DIAP1 and removal of DIAP1 permits activation of DRONC by the Drosophila Apaf-1-related killer, ARK. ARK is proposed to facilitate DRONC oligomerisation and autoprocessing at residue E352. This study examines whether autoprocessing of DRONC is required for its activation and for DRONC-mediated cell death. Using purified recombinant proteins, we show here that while DRONC autocleaves at residue E352, mutation of this site did not abolish enzyme activation, DRICE-induced cleavage of DRONC or DRONC-mediated activation of DRICE. We performed a detailed mutational analysis of DRONC cleavage sites and show that overexpression of DRONC cleavage mutants in Drosophila cells retain pro-apoptotic activity. Using an in vitro cell-free assay, we found ARK alone did not activate DRONC and demonstrate a requirement for an additional cytosolic factor in ARK-mediated DRONC activation. These results suggest that, similar to mammalian caspase-2 and caspase-9, the initial cleavage of DRONC is not essential for its activation and suggest a mechanism of ARK-mediated DRONC activation different from that proposed previously.
Publisher: Springer Science and Business Media LLC
Date: 26-04-2013
DOI: 10.1038/CDD.2013.35
Publisher: Elsevier BV
Date: 02-2015
Publisher: Springer Science and Business Media LLC
Date: 05-01-2007
Abstract: The Bcl-2-family of proteins localize to intracellular membranes via a C-terminal hydrophobic membrane anchor (MA) domain, to exert their antiapoptotic or proapoptotic functions. In Drosophila, both Bcl-2 family members, DEBCL and BUFFY, contain an MA. In DEBCL the MA is necessary for the localization of protein to mitochondria and for its proapoptotic activity. BUFFY is highly similar to DEBCL but its localization and function are not clearly defined. Here, we report on comparative analysis of BUFFY and DEBCL to decipher the molecular basis for their subcellular localization. We show that these two proteins localize to distinct intracellular membranes, DEBCL predominantly to mitochondria and BUFFY to endoplasmic reticula (ER). Our results suggest that the MA-flanking residues in DEBCL, homologous to Bcl-X(L), are required for the targeting of DEBCL to mitochondria. The C-terminal positively charged residues present in DEBCL are absent in BUFFY, which allows for its localization to ER. The MA in both proteins is required for the correct targeting and proapoptotic activities of these proteins. Interestingly, a functional nuclear localization signal was identified in the N-terminal region of BUFFY and in the absence of the MA, BUFFY accumulated in the nucleus. The functional implications of these findings are discussed.
Publisher: Proceedings of the National Academy of Sciences
Date: 31-03-2009
Abstract: Apoptosis is mediated by the caspase family of proteases that act as effectors of cell death by cleaving many cellular substrates. Caspase-2 is one of the most evolutionarily conserved caspases, yet its physiological function has remained enigmatic because caspase-2-deficient mice develop normally and are viable. We report here that the caspase-2 −/− mouse embryonic fibroblasts (MEFs) show increased proliferation. When transformed with E1A and Ras oncogenes, caspase-2 −/− MEFs grew significantly faster than caspase-2 +/+ MEFs and formed more aggressive and accelerated tumors in nude mice. To assess whether the loss of caspase-2 predisposes animals to tumor development, we used the mouse Eμ- Myc lymphoma model. Our findings suggest that loss of even a single allele of caspase-2 resulted in accelerated tumorigenesis, and this was further enhanced in caspase-2 −/− mice. The caspase-2 −/− cells showed resistance to apoptosis induced by chemotherapeutic drugs and DNA damage. Furthermore, caspase-2 −/− MEFs had a defective apoptotic response to cell-cycle checkpoint regulation and showed abnormal cycling following γ-irradiation. These data show that loss of caspase-2 results in an increased ability of cells to acquire a transformed phenotype and become malignant, indicating that caspase-2 is a tumor suppressor protein.
Publisher: Springer Science and Business Media LLC
Date: 15-05-2018
Publisher: Springer Science and Business Media LLC
Date: 22-01-2015
Abstract: Ageing is a complex biological process for which underlying biochemical changes are still largely unknown. We performed comparative profiling of the cellular proteome and metabolome to understand the molecular basis of ageing in Caspase-2- deficient ( Casp2 −/− ) mice that are a model of premature ageing in the absence of overt disease. Age-related changes were determined in the liver and serum of young (6–9 week) and aged (18–24 month) wild-type and Casp2 −/− mice. We identified perturbed metabolic pathways, decreased levels of ribosomal and respiratory complex proteins and altered mitochondrial function that contribute to premature ageing in the Casp2 −/− mice. We show that the metabolic profile changes in the young Casp2 −/− mice resemble those found in aged wild-type mice. Intriguingly, aged Casp2 −/− mice were found to have reduced blood glucose and improved glucose tolerance. These results demonstrate an important role for caspase-2 in regulating proteome and metabolome remodelling during ageing.
Publisher: Springer Science and Business Media LLC
Date: 22-12-2014
DOI: 10.1038/ONC.2014.413
Abstract: Caspase-2 belongs to the caspase family of cysteine proteases with established roles in apoptosis. Recently, caspase-2 has been implicated in nonapoptotic functions including maintenance of genomic stability and tumor suppression. Our previous studies demonstrated that caspase-2 also regulates cellular redox status and delays the onset of several ageing-related traits. In the current study, we tested stress tolerance ability in caspase-2-deficient (Casp2(-/-)) mice by challenging both young and old mice with a low dose of the potent reactive oxygen species (ROS) generator, PQ that primarily affects lungs. In both groups of mice, PQ induced pulmonary damage. However, the lesions in caspase-2 knockout mice were consistently and reproducibly more severe than those in wild-type (WT) mice. Furthermore, serum interleukin (IL)-1β and IL-6 levels were higher in PQ-exposed aged Casp2(-/-) mice indicating increased inflammation. Interestingly, livers from Casp2(-/-) mice displayed karyomegaly, a feature commonly associated with ageing and aneuploidy. Given that Casp2(-/-) mice show impaired antioxidant defense, we tested oxidative damage in these mice. Protein oxidation significantly increased in PQ-injected old Casp2(-/-) mice. Moreover, FoxO1, SOD2 and Nrf2 expression levels were reduced and induction of superoxide dismutase (SOD) and glutathione peroxidase activity was not observed in PQ-treated Casp2(-/-) mice. Strong c-Jun amino-terminal kinase (JNK) activation was observed in Casp2(-/-) mice, indicative of increased stress. Together, our data strongly suggest that caspase-2 deficiency leads to increased cellular stress largely because these mice fail to respond to oxidative stress by upregulating their antioxidant defense mechanism. This makes the mice more vulnerable to exogenous challenges and may partly explain the shorter lifespan of Casp2(-/-) mice.
Publisher: Springer Science and Business Media LLC
Date: 21-08-2014
Publisher: Springer Science and Business Media LLC
Date: 06-08-2018
Publisher: Wiley
Date: 08-12-2003
Publisher: Springer Science and Business Media LLC
Date: 10-1997
Publisher: Proceedings of the National Academy of Sciences
Date: 18-11-2013
Abstract: The cysteine protease caspase-2 has been implicated in the suppression of oncogene-mediated tumor formation. However, the mechanisms underlying the function of caspase-2 as a tumor suppressor are not well defined. In this study, we use a well-characterized mouse lymphoma model and demonstrate a critical role for caspase-2 in maintaining genome stability and in the suppression of tumorigenesis following loss of the essential DNA repair gene ataxia telangiectasia mutated ( Atm ). Our findings suggest that caspase-2 cooperates with ATM to suppress genomic instability, oxidative stress, and tumor progression.
Publisher: Humana Press
Date: 2009
DOI: 10.1007/978-1-60327-017-5_1
Abstract: Apoptotic cell death is characterised by various morphological and biochemical changes. Cysteine proteases of the caspase family play key roles in the execution of apoptosis and in the maturation of proinflammatory cytokines. During apoptosis signalling, caspase precursors undergo rapid proteolytic processing and activation. Activated caspases then function to cleave various vital cellular proteins, resulting in the death of the cell. Thus, the measurement of caspase activation and caspase activity provides a quick and convenient method to assess apoptosis. This chapter outlines various commonly used assays for measuring caspase activity and detecting active caspases in cultured cells or tissue extracts.
Publisher: Springer Science and Business Media LLC
Date: 07-04-2006
Publisher: Proceedings of the National Academy of Sciences
Date: 13-04-1999
Abstract: Caspases play an essential role in the execution of programmed cell death in metazoans. Although 14 caspases are known in mammals, only a few have been described in other organisms. Here we describe the identification and characterization of a Drosophila caspase, DRONC, that contains an amino terminal caspase recruitment domain. Ectopic expression of DRONC in cultured cells resulted in apoptosis, which was inhibited by the caspase inhibitors p35 and MIHA. DRONC exhibited a substrate specificity similar to mammalian caspase-2. DRONC is ubiquitously expressed in Drosophila embryos during early stages of development. In late third instar larvae, dronc mRNA is dramatically up-regulated in salivary glands and midgut before histolysis of these tissues. Exposure of salivary glands and midgut isolated from second instar larvae to ecdysone resulted in a massive increase in dronc mRNA levels. These results suggest that DRONC is an effector of steroid-mediated apoptosis during insect metamorphosis.
Publisher: Springer New York
Date: 2014
DOI: 10.1007/978-1-4939-0357-3_4
Abstract: Caspase-2 has been shown to function in apoptosis and in some non-apoptotic pathways, including tumor suppression and aging. Caspase-2 has some unique features and is the only caspase that constitutively localizes to the nucleus, although its nuclear function remains unknown. During apoptosis signaling, caspase-2 rapidly homodimerizes, which leads to its activation and proteolytic processing. The activation of caspase-2 can be measured by assessing its dimerization and/or cleavage of the caspase-2 zymogen and its substrates. This chapter outlines commonly used methods to purify recombinant caspase-2 and assess its activity and function in vitro and in cultured cells or tissue extracts.
Publisher: Springer Science and Business Media LLC
Date: 26-10-2017
Abstract: Caspase-2 has been shown to be involved in metabolic homeostasis. Here, we show that caspase-2 deficiency alters basal energy metabolism by shifting the balance in fuel choice from fatty acid to carbohydrate usage. At 4 weeks of age, whole-body carbohydrate utilisation was increased in Casp2 −/− mice and was maintained into adulthood. By 17 weeks of age, Casp2 −/− mice had reduced white adipose mass, smaller white adipocytes decreased fasting blood glucose and plasma triglycerides but maintained normal insulin levels. When placed on a 12-week high-fat diet (HFD), Casp2 −/− mice resisted the development of obesity, fatty liver, hyperinsulinemia and insulin resistance. In addition, HFD-fed Casp2 −/− mice had reduced white adipocyte hypertrophy, apoptosis and expansion of both subcutaneous and visceral adipose depots. Increased expression of UCP1 and the maintenance of adiponectin levels in white adipose tissue of HFD-fed Casp2 −/− mice indicated increased browning and adipocyte hyperplasia. We found that while the preference for whole-body carbohydrate utilisation was maintained, HFD-fed Casp2 −/− mice were not impaired in their ability to switch to utilising fats as a fuel source. Our findings suggest that caspase-2 impacts basal energy metabolism by regulating adipocyte biology and fat expansion, most likely via a non-apoptotic function. Furthermore, we show that caspase-2 deficiency shifts the balance in fuel choice towards increased carbohydrate utilisation and propose that this is due to mild energy stress. As a consequence, Casp2 −/− mice show an adaptive remodelling of adipose tissue that protects from HFD-induced obesity and improves glucose homeostasis while paradoxically increasing their susceptibility to oxidative stress induced damage and premature ageing.
Publisher: Elsevier BV
Date: 10-1999
Publisher: Elsevier BV
Date: 12-2000
Publisher: Springer Science and Business Media LLC
Date: 14-01-2008
Abstract: Caspase-2 is one of the most conserved caspases, yet its biological function remains a matter of controversy. In the present article we analysed mouse embryonic fibroblasts (MEFs) from caspase-2 knockout mice for their sensitivity to various apoptosis inducing agents. We found that cell death induced by drugs that disrupt cytoskeleton is significantly inhibited in Casp2(-/-) MEFs. These drugs included zoledronic acid, vincristine, cytochalasin D and paclitaxel. We demonstrate that MEFs lacking Casp2 show clonogenic survival following drug treatment, whereas all Casp2(+/+) MEFs die, indicating that caspase-2 is required for apoptosis induced by cytoskeletal disruption. We further found that caspase-2 mediates apoptosis via Piddosome, Bid and Bax activation, and cytochrome c release. In the absence of caspase-2, Bid and Bax activation, and cytochrome c release are significantly delayed following drug treatment. Our data provide strong support for a context-dependent function of caspase-2 in apoptosis.
Publisher: Rockefeller University Press
Date: 08-11-2004
Abstract: In Drosophila, activation of the apical caspase DRONC requires the apoptotic protease-activating factor homologue, DARK. However, unlike caspase activation in mammals, DRONC activation is not accompanied by the release of cytochrome c from mitochondria. Drosophila encodes two cytochrome c proteins, Cytc-p (DC4) the predominantly expressed species, and Cytc-d (DC3), which is implicated in caspase activation during spermatogenesis. Here, we report that silencing expression of either or both DC3 and DC4 had no effect on apoptosis or activation of DRONC and DRICE in Drosophila cells. We find that loss of function mutations in dc3 and dc4, do not affect caspase activation during Drosophila development and that ectopic expression of DC3 or DC4 in Drosophila cells does not induce caspase activation. In cell-free studies, recombinant DC3 or DC4 failed to activate caspases in Drosophila cell lysates, but remarkably induced caspase activation in extracts from human cells. Overall, our results argue that DARK-mediated DRONC activation occurs independently of cytochrome c.
Publisher: Springer Science and Business Media LLC
Date: 12-2016
Abstract: The apoptotic cysteine protease caspase-2 has been shown to suppress tumourigenesis in mice and its reduced expression correlates with poor prognosis in some human malignancies. Caspase-2-deficient mice develop normally but show ageing-related traits and, when challenged by oncogenic stimuli or certain stress, show enhanced tumour development, often accompanied by extensive aneuploidy. As stem cells are susceptible to acquiring age-related functional defects because of their self-renewal and proliferative capacity, we examined whether loss of caspase-2 promotes such defects with age. Using young and aged Casp2 −/− mice, we demonstrate that deficiency of caspase-2 results in enhanced aneuploidy and DNA damage in bone marrow (BM) cells with ageing. Furthermore, we demonstrate for the first time that caspase-2 loss results in significant increase in immunophenotypically defined short-term haematopoietic stem cells (HSCs) and multipotent progenitors fractions in BM with a skewed differentiation towards myeloid progenitors with ageing. Caspase-2 deficiency leads to enhanced granulocyte macrophage and erythroid progenitors in aged mice. Colony-forming assays and long-term culture-initiating assay further recapitulated these results. Our results provide the first evidence of caspase-2 in regulating HSC and progenitor differentiation, as well as aneuploidy, in vivo .
Publisher: Rockefeller University Press
Date: 18-03-2002
Abstract: The release of cytochrome c from mitochondria is necessary for the formation of the Apaf-1 apoptosome and subsequent activation of caspase-9 in mammalian cells. However, the role of cytochrome c in caspase activation in Drosophila cells is not well understood. We demonstrate here that cytochrome c remains associated with mitochondria during apoptosis of Drosophila cells and that the initiator caspase DRONC and effector caspase DRICE are activated after various death stimuli without any significant release of cytochrome c in the cytosol. Ectopic expression of the proapoptotic Bcl-2 protein, DEBCL, also fails to show any cytochrome c release from mitochondria. A significant proportion of cellular DRONC and DRICE appears to localize near mitochondria, suggesting that an apoptosome may form in the vicinity of mitochondria in the absence of cytochrome c release. In vitro, DRONC was recruited to a & -kD complex, similar to the mammalian apoptosome in cell extracts supplemented with cytochrome c and dATP. These results suggest that caspase activation in insects follows a more primitive mechanism that may be the precursor to the caspase activation pathways in mammals.
Publisher: Springer Science and Business Media LLC
Date: 17-02-2012
DOI: 10.1038/CDD.2012.13
Publisher: Springer Science and Business Media LLC
Date: 13-04-2012
DOI: 10.1038/CDD.2012.36
Publisher: Springer Science and Business Media LLC
Date: 28-06-2013
DOI: 10.1038/CDD.2013.87
Publisher: Proceedings of the National Academy of Sciences
Date: 08-09-2009
Abstract: The regulation of metal ion transport within neurons is critical for normal brain function. Of particular importance is the regulation of redox metals such as iron (Fe), where excess levels can contribute to oxidative stress and protein aggregation, leading to neuronal death. The alent metal transporter 1 (DMT1) plays a central role in the regulation of Fe as well as other metals hence, failure of DMT1 regulation is linked to human brain pathology. However, it remains unclear how DMT1 is regulated in the brain. Here, we show that DMT1 is regulated by Ndfip1 (Nedd4 family-interacting protein 1), an adaptor protein that recruits E3 ligases to ubiquitinate target proteins. Using human neurons we show the Ndfip1 is upregulated and binds to DMT1 in response to Fe and cobalt (Co) exposure. This interaction results in the ubiquitination and degradation of DMT1, resulting in reduced metal entry. Induction of Ndfip1 expression protects neurons from metal toxicity, and removal of Ndfip1 by shRNAi results in hypersensitivity to metals. We identify Nedd4–2 as an E3 ligase recruited by Ndfip1 for the ubiquitination of DMT1 within human neurons. Comparison of brains from Ndfip1 −/− with Ndfip1 +/+ mice exposed to Fe reveals that Ndfip1 −/− brains accumulate Fe within neurons. Together, this evidence suggests a critical role for Ndfip1 in regulating metal transport in human neurons.
Publisher: Faculty Opinions Ltd
Date: 15-12-2009
DOI: 10.3410/B1-96
Publisher: Springer Science and Business Media LLC
Date: 19-12-2017
DOI: 10.1038/ONC.2016.423
Publisher: Springer Science and Business Media LLC
Date: 22-01-2019
DOI: 10.1038/S41419-018-1296-0
Abstract: Caspase-2 is a highly conserved cysteine protease with roles in apoptosis and tumor suppression. Our recent findings have also demonstrated that the tumor suppression function of caspase-2 is context specific. In particular, while caspase-2 deficiency augments lymphoma development in the EμMyc mouse model, it leads to delayed neuroblastoma development in Th-MYCN mice. However, it is unclear how caspase-2 mediates these differential outcomes. Here we utilized RNA sequencing to define the transcriptomic changes caused by caspase-2 ( Casp2 −/− ) deficiency in tumors from EμMyc and Th-MYCN mice. We describe key changes in both lymphoma and neuroblastoma-associated genes and identified differential expression of the EGF-like domain-containing gene, Megf6 , in the two tumor types that may contribute to tumor outcome following loss of Casp2 . We identified a panel of genes with altered expression in Th-MYCN/Casp2 −/− tumors that are strongly associated with neuroblastoma outcome, with roles in melanogenesis, Wnt and Hippo pathway signaling, that also contribute to neuronal differentiation. In contrast, we found that key changes in gene expression in the EμMyc/Casp2 −/− tumors, are associated with increased immune signaling and T-cell infiltration previously associated with more aggressive lymphoma progression. In addition, Rap1 signaling pathway was uniquely enriched in Casp2 deficient EμMyc tumors. Our findings suggest that Casp2 deficiency augments immune signaling pathways that may be in turn, enhance lymphomagenesis. Overall, our study has identified new genes and pathways that contribute to the caspase-2 tumor suppressor function and highlight distinct roles for caspase-2 in different tissues.
Publisher: American Society of Hematology
Date: 15-11-2008
DOI: 10.1182/BLOOD-2008-04-150953
Abstract: Many ion channels and transporters are regulated by ubiquitination mediated by the Nedd4 family of HECT-type ubiquitin ligases (E3s). These E3s commonly interact with substrates via their WW domains that bind to specific motifs in target proteins. However, not all potential targets of these E3s contain WW-binding motifs. Therefore, accessory proteins may mediate the interaction between Nedd4 family members and their targets. Here we report that the alent metal ion transporter DMT1, the primary nonheme iron transporter in mammals, is regulated by ubiquitination mediated by the Nedd4 family member WWP2. DMT1 interacts with 2 WW domain-interacting proteins, Ndfip1 and Ndfip2, previously proposed to have roles in protein trafficking. This promotes DMT1 ubiquitination and degradation by WWP2. Consistent with these observations, Ndfip1−/− mice show increased DMT1 activity and a concomitant increase in hepatic iron deposition, indicating an essential function of Ndfip1 in iron homeostasis. This novel mechanism of regulating iron homeostasis suggests that Ndfips and WWP2 may contribute to diseases involving aberrant iron transport.
Publisher: Springer Science and Business Media LLC
Date: 12-08-2016
DOI: 10.1038/CDD.2016.81
Publisher: Springer Science and Business Media LLC
Date: 29-11-2014
DOI: 10.1038/CDD.2013.168
Publisher: Springer Science and Business Media LLC
Date: 19-12-2015
DOI: 10.1038/CDD.2014.216
Publisher: Elsevier BV
Date: 2001
Publisher: Elsevier BV
Date: 2011
No related grants have been discovered for Loretta Dorstyn.