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HIPK2–P53ser46 Damage Response Pathway Is Involved in Temozolomide-Induced Glioblastoma Cell Death Yang He, Wynand P
Published OnlineFirst February 22, 2019; DOI: 10.1158/1541-7786.MCR-18-1306 Cell Fate Decisions Molecular Cancer Research The SIAH1–HIPK2–p53ser46 Damage Response Pathway is Involved in Temozolomide-Induced Glioblastoma Cell Death Yang He, Wynand P. Roos, Qianchao Wu, Thomas G. Hofmann, and Bernd Kaina Abstract Patients suffering from glioblastoma have a dismal prog- in chromatin-immunoprecipitation experiments, in which nosis, indicating the need for new therapeutic targets. Here p-p53ser46 binding to the Fas promotor was regulated by we provide evidence that the DNA damage kinase HIPK2 HIPK2. Other pro-apoptotic proteins such as PUMA, and its negative regulatory E3-ubiquitin ligase SIAH1 are NOXA, BAX, and PTEN were not affected in HIPK2kd, and critical factors controlling temozolomide-induced cell also double-strand breaks following temozolomide remain- death. We show that HIPK2 downregulation (HIPK2kd) ed unaffected. We further show that downregulation of significantly reduces the level of apoptosis. This was not the HIPK2 inactivator SIAH1 significantly ameliorates the case in glioblastoma cells expressing the repair protein temozolomide-induced apoptosis, suggesting that the MGMT, suggesting that the primary DNA lesion responsible ATM/ATR target SIAH1 together with HIPK2 plays a pro- 6 for triggering HIPK2-mediated apoptosis is O -methylguanine. apoptotic role in glioma cells exhibiting p53wt status. A Upon temozolomide treatment, p53 becomes phosphory- database analysis revealed that SIAH1, but not SIAH2, is lated whereby HIPK2kd had impact exclusively on ser46, significantly overexpressed in glioblastomas. but not ser15. Searching for the transcriptional target of p-p53ser46, we identified the death receptor FAS (CD95, Implications: The identification of a novel apoptotic APO-1) being involved. -
Critical Function of Siah2 in Tumorigenesis
AIMS Molecular Science, 4(4): 415-423 DOI: 10.3934/molsci.2017.4.415 Received 31 July 2017, Accepted 21 September 2017, Published 11 October 2017 http://www.aimspress.com/journal/Molecular Review Critical function of Siah2 in tumorigenesis Kazunobu Baba, Tadaaki Miyazaki* Department of Probiotics Immunology, Institute for Genetic Medicine, Hokkaido University, North 15 West 7, Kita-ku, Sapporo City, Hokkaido, Japan * Correspondence: Email: [email protected]; Tel: 81-011-706-8095. Abstract: The seven in absentia homolog (Siah) family proteins are components of E3 RING zinc finger ubiquitin ligase complexes that catalyze the ubiquitination of proteins. Siah proteins target their substrates for proteasomal degradation. Evidence is growing that Siah proteins are implicated in the progression of various cancer cells and play a critical role in angiogenesis and tumorigenesis, particularly through Ras, p53, estrogen, and hypoxia inducible factor (HIF)-mediated signaling pathways in response to DNA damage or hypoxia. Keywords: Siah2; Nrf2; ROS metabolism; Hypoxia 1. Introduction The seven in absentia (Sina) gene was initially identified to be critical for photoreceptor cell development in Drosophila [1]. As its mammalian orthologs, murine and human homologs of the Sina gene have also been identified. In Xenopus and mice, Sina homologs (Siahs) were isolated as three Siah proteins, Siah1a, Siah1b, and Siah2, which are encoded by different genes [2]. In contrast, the human Siah proteins consist of two homologs, Siah1 and Siah2, which are encoded by the SIAH1 and SIAH2 genes, respectively [3]. Siah1 and Siah2 have the high sequence similarity of their N-terminal RING finger domain, central cysteine-rich domain, and C-terminal substrate binding domain (SBD); however, Siah1 and Siah2 apparently have distinct but overlapping functions as proteins of a tumor suppressor gene and a proto-oncogene, respectively (Figure 1) [4]. -
Is Glyceraldehyde-3-Phosphate Dehydrogenase a Central Redox Mediator?
1 Is glyceraldehyde-3-phosphate dehydrogenase a central redox mediator? 2 Grace Russell, David Veal, John T. Hancock* 3 Department of Applied Sciences, University of the West of England, Bristol, 4 UK. 5 *Correspondence: 6 Prof. John T. Hancock 7 Faculty of Health and Applied Sciences, 8 University of the West of England, Bristol, BS16 1QY, UK. 9 [email protected] 10 11 SHORT TITLE | Redox and GAPDH 12 13 ABSTRACT 14 D-Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) is an immensely important 15 enzyme carrying out a vital step in glycolysis and is found in all living organisms. 16 Although there are several isoforms identified in many species, it is now recognized 17 that cytosolic GAPDH has numerous moonlighting roles and is found in a variety of 18 intracellular locations, but also is associated with external membranes and the 19 extracellular environment. The switch of GAPDH function, from what would be 20 considered as its main metabolic role, to its alternate activities, is often under the 21 influence of redox active compounds. Reactive oxygen species (ROS), such as 22 hydrogen peroxide, along with reactive nitrogen species (RNS), such as nitric oxide, 23 are produced by a variety of mechanisms in cells, including from metabolic 24 processes, with their accumulation in cells being dramatically increased under stress 25 conditions. Overall, such reactive compounds contribute to the redox signaling of the 26 cell. Commonly redox signaling leads to post-translational modification of proteins, 27 often on the thiol groups of cysteine residues. In GAPDH the active site cysteine can 28 be modified in a variety of ways, but of pertinence, can be altered by both ROS and 29 RNS, as well as hydrogen sulfide and glutathione. -
Siah - a Promising Anti-Cancer Target
Author Manuscript Published OnlineFirst on March 1, 2013; DOI: 10.1158/0008-5472.CAN-12-4348 Author manuscripts have been peer reviewed and accepted for publication but have not yet been edited. Siah - a promising anti-cancer target Christina SF Wong1 and Andreas Möller1 1 Tumour Microenvironment Laboratory, Queensland Institute of Medical Research, 300 Herston Road, Herston, Queensland 4006, Australia. Corresponding Author: Andreas Möller ([email protected]) Running title: Siah and Cancer Keywords: Siah1, Siah2, E3 Ubiquitin ligases, Cancer Potential conflict of interest: The authors declare no conflict of interest. Word count: Abstract: 100 words; Text: 2590 words; Number of Figures: 1; Number of Tables: 1 1 Downloaded from cancerres.aacrjournals.org on September 29, 2021. © 2013 American Association for Cancer Research. Author Manuscript Published OnlineFirst on March 1, 2013; DOI: 10.1158/0008-5472.CAN-12-4348 Author manuscripts have been peer reviewed and accepted for publication but have not yet been edited. Abstract: Siah ubiquitin ligases play important roles in a number of signaling pathways involved in the progression and spread of cancer in cell-based models but their role in tumor progression remains controversial. Siah proteins have been described to be both oncogenic as well as tumor-suppressive in a variety of patient cohort studies and animal cancer models. This review collates the current knowledge of Siah in cancer progression and identifies potential methods of translation of these findings into the clinic. Furthermore, key experiments needed to close the gaps in our understanding of the role Siah proteins play in tumor progression are suggested. 2 Downloaded from cancerres.aacrjournals.org on September 29, 2021. -
SIAH2-Mediated and Organ-Specific Restriction of HO-1 Expression by A
www.nature.com/scientificreports OPEN SIAH2-mediated and organ-specifc restriction of HO-1 expression by a dual mechanism Shashipavan Chillappagari1*, Ratnal Belapurkar1, Andreas Möller2, Nicole Molenda3, Michael Kracht4, Susanne Rohrbach3 & M. Lienhard Schmitz1* The intracellular levels of the cytoprotective enzyme heme oxygenase-1 (HO-1) are tightly controlled. Here, we reveal a novel mechanism preventing the exaggerated expression of HO-1. The analysis of mice with a knock-out in the ubiquitin E3 ligase seven in absentia homolog 2 (SIAH2) showed elevated HO-1 protein levels in specifc organs such as heart, kidney and skeletal muscle. Increased HO-1 protein amounts were also seen in human cells deleted for the SIAH2 gene. The higher HO-1 levels are not only due to an increased protein stability but also to elevated expression of the HO-1 encoding HMOX1 gene, which depends on the transcription factor nuclear factor E2-related factor 2 (NRF2), a known SIAH2 target. Dependent on its RING (really interesting new gene) domain, expression of SIAH2 mediates proteasome-dependent degradation of its interaction partner HO-1. Additionally SIAH2-defcient cells are also characterized by reduced expression levels of glutathione peroxidase 4 (GPX4), rendering the knock-out cells more sensitive to ferroptosis. Ubiquitin E3 ligases regulate the activity and turnover of many target proteins, thus controlling key features such as metabolism, stress signaling and cell cycle progression1. Te RING family of ubiquitin E3 ligases comprises the SIAH family. Te human genome encodes SIAH1 and the homologous SIAH2 protein as well as SIAH3, which lacks a functional RING domain and is only expressed in a limited subset of cancer cell lines2. -
TRB3 Is Elevated in Psoriasis Vulgaris Lesions and Mediates Hacat Cells Proliferation in Vitro Xiao-Jing Yu, Tie-Jun Song, Lu-Wei Zhang, Ying Su, Ke-Yu Wang, Qing Sun
Brief report J Investig Med: first published as 10.1136/jim-2017-000453 on 7 August 2017. Downloaded from TRB3 is elevated in psoriasis vulgaris lesions and mediates HaCaT cells proliferation in vitro Xiao-Jing Yu, Tie-Jun Song, Lu-Wei Zhang, Ying Su, Ke-Yu Wang, Qing Sun Department of Dermatology, ABSTRACT It presents clinically as a sharply demarcated Qilu Hospital of Shandong Psoriasis is a chronic skin disease characterized erythematous plaque with silvery white scales. University, Jinan, Shandong, Histologically, it is characterized by hyperkera- China by abnormal keratinocyte proliferation and differentiation, inflammation, and angiogenesis. tosis, parakeratosis, acanthosis of the epidermis, Correspondence to Overexpression of tribbles homolog3 (TRB3), which tortuous and dilated vessels, and an inflamma- Dr Qing Sun, Department of belongs to the tribbles family of pseudokinases, has tory infiltrate composed mostly of lymphocytes. Dermatology, Qilu Hospital, been found in several human tumors and metabolic The pathogenesis of psoriasis is complex and the University of Shandong, diseases, but its role in psoriasis has not been fully exact mechanism remains elusive, but abnormal Jinan 250012, China; proliferation of keratinocytes is an important sunqing7226@ 163. com clarified. The aim of this study is to investigate the expression of TRB3 in psoriasis and explore its roles mechanism of psoriasis, resulting in epidermal Received 25 February 2017 in the proliferation of keratinocytes. Twenty-four hyperplasia and a morphologic characteristic of Revised 28 June 2017 patients with psoriasis vulgaris were recruited for the psoriasis.16 Numerous well established antipso- Accepted 2 July 2017 study. Diagnosis of psoriasis was based on clinical riatic treatments, including phototherapy, oral Published Online First 7 August 2017 and histologic examinations. -
Rabbit Anti-SIAH1/FITC Conjugated Antibody-SL3596R-FITC
SunLong Biotech Co.,LTD Tel: 0086-571- 56623320 Fax:0086-571- 56623318 E-mail:[email protected] www.sunlongbiotech.com Rabbit Anti-SIAH1/FITC Conjugated antibody SL3596R-FITC Product Name: Anti-SIAH1/FITC Chinese Name: FITC标记的Ubiquitin连接酶Siah1抗体 hSIAH1; HUMSIAH; Seven in absentia homolog 1 (Drosophila); Seven in absentia homolog 1; Siah 1; Siah 1a; Ubiquitin ligase SIAH1; E3 ubiquitin-protein ligase SIAH1; Alias: Seven in absentia homolog 1; Siah-1; Siah-1a; Siah E3 ubiquitin protein ligase 1; SIAH1_HUMAN; SIAH1A. Organism Species: Rabbit Clonality: Polyclonal React Species: Human,Mouse,Rat,Chicken,Dog,Pig,Cow,Horse,Rabbit, IF=1:50-200 Applications: not yet tested in other applications. optimal dilutions/concentrations should be determined by the end user. Molecular weight: 34kDa Form: Lyophilized or Liquid Concentration: 1mg/ml immunogen: KLH conjugated synthetic peptide derived from human SIAH1 Lsotype: IgG Purification: affinitywww.sunlongbiotech.com purified by Protein A Storage Buffer: 0.01M TBS(pH7.4) with 1% BSA, 0.03% Proclin300 and 50% Glycerol. Store at -20 °C for one year. Avoid repeated freeze/thaw cycles. The lyophilized antibody is stable at room temperature for at least one month and for greater than a year Storage: when kept at -20°C. When reconstituted in sterile pH 7.4 0.01M PBS or diluent of antibody the antibody is stable for at least two weeks at 2-4 °C. background: This gene encodes a protein that is a member of the seven in absentia homolog (SIAH) family. The protein is an E3 ligase and is involved in ubiquitination and proteasome- Product Detail: mediated degradation of specific proteins. -
SIAH1-Mediated RPS3 Ubiquitination Contributes to Chemosensitivity in Epithelial Ovarian Cancer
SIAH1-Mediated RPS3 Ubiquitination Contributes to Chemosensitivity in Epithelial Ovarian Cancer Lu Chen Zhenjiang Fourth Peoples Hospital and Zhenjiang Women and Childrens Hospital Wujiang Gao Zhenjiang Fourth Peoples Hospital and Zhenjiang Women and Childrens Hospital Chunli Sha Zhenjiang Fourth Peoples Hospital and Zhenjiang Women and Childrens Hospital Taoqiong Li Zhenjiang Fourth Peoples Hospital and Zhenjiang Women and Childrens Hospital Li Lin Zhenjiang Fourth Peoples Hospital and Zhenjiang Women and Childrens Hospital Hong Wei Zhenjiang Fourth Peoples Hospital and Zhenjiang Women and Childrens Hospital Qi Chen Zhenjiang Fourth Peoples Hospital and Zhenjiang Women and Childrens Hospital Jie Xing Zhenjiang Fourth Peoples Hospital and Zhenjiang Women and Childrens Hospital Mengxue Zhang Zhenjiang Fourth Peoples Hospital and Zhenjiang Women and Childrens Hospital Shijie Zhao Zhenjiang Fourth Peoples Hospital and Zhenjiang Women and Childrens Hospital Wenlin Xu Zhenjiang Fourth Peoples Hospital and Zhenjiang Women and Childrens Hospital Yuefeng Li Jiangsu University Hospital Xiaolan Zhu ( [email protected] ) Zhenjiang Fourth Peoples Hospital and Zhenjiang Women and Childrens Hospital Research Keywords: EOC, chemoresistance, SIAH1, RPS3,NF-κB Posted Date: July 20th, 2021 Page 1/27 DOI: https://doi.org/10.21203/rs.3.rs-697379/v1 License: This work is licensed under a Creative Commons Attribution 4.0 International License. Read Full License Page 2/27 Abstract Background: The E3 ligase SIAH1 is deregulated in human cancers and correlated with poor prognosis, but its contributions to chemoresistance in epithelial ovarian cancer (EOC) are not evident. Methods: SIAHI, RPS3, NF-κB (P65), FLAG and GFP protein levels were assessed by western blotting (WB) or immunohistochemistry (IHC). The mRNA levels were revealed by qRT-PCR. -
Loss of HIPK2 Protects Neurons from Mitochondrial Toxins by Regulating Parkin Protein Turnover
The Journal of Neuroscience, January 15, 2020 • 40(3):557–568 • 557 Cellular/Molecular Loss of HIPK2 Protects Neurons from Mitochondrial Toxins by Regulating Parkin Protein Turnover X Jiasheng Zhang,1,2 Yulei Shang,1 Sherry Kamiya,1 Sarah J. Kotowski,3,4 XKen Nakamura,3,4 and XEric J. Huang1,2 1Department of Pathology, University of California, San Francisco, San Francisco, California 94143, 2Pathology Service 113B, VA Medical Center, San Francisco, California 94121, 3Department of Neurology, University of California, San Francisco, San Francisco, California 94122, and 4Gladstone Institute of Neurological Disease, San Francisco, California 94158 Mitochondria are important sources of energy, but they are also the target of cellular stress, toxin exposure, and aging-related injury. Persistent accumulation of damaged mitochondria has been implicated in many neurodegenerative diseases. One highly conserved mechanism to clear damaged mitochondria involves the E3 ubiquitin ligase Parkin and PTEN-induced kinase 1 (PINK1), which cooper- atively initiate the process called mitophagy that identifies and eliminates damaged mitochondria through the autophagosome and lysosome pathways. Parkin is a mostly cytosolic protein, but is rapidly recruited to damaged mitochondria and target them for mi- tophagy. Moreover, Parkin interactomes also involve signaling pathways and transcriptional machinery critical for survival and cell death. However, the mechanism that regulates Parkin protein level remains poorly understood. Here, we show that the loss of homeodo- main interacting protein kinase 2 (HIPK2) in neurons and mouse embryonic fibroblasts (MEFs) has a broad protective effect from cell death induced by mitochondrial toxins. The mechanism by which Hipk2 Ϫ / Ϫ neurons and MEFs are more resistant to mitochondrial toxins is in part due to the role of HIPK2 and its kinase activity in promoting Parkin degradation via the proteasome-mediated mecha- nism. -
PHF19 Promotes the Proliferation, Migration, and Chemosensitivity Of
Deng et al. Cell Death and Disease (2018) 9:1049 DOI 10.1038/s41419-018-1082-z Cell Death & Disease ARTICLE Open Access PHF19 promotes the proliferation, migration, and chemosensitivity of glioblastoma to doxorubicin through modulation of the SIAH1/β–catenin axis Qing Deng1, Jianbing Hou1, Liying Feng1, Ailing Lv1, Xiaoxue Ke1, Hanghua Liang1,FengWang1,KuiZhang1, Kuijun Chen2 and Hongjuan Cui1 Abstract PHD finger protein 19 (PHF19), a critical component of the polycomb repressive complex 2 (PRC2), is crucial for maintaining the repressive transcriptional activity of several developmental regulatory genes and plays essential roles in various biological processes. Abnormal expression of PHF19 causes dysplasia or serious diseases, including chronic myeloid disorders and tumors. However, the biological functions and molecular mechanisms of PHF19 in glioblastoma (GBM) remain unclear. Here, we demonstrated that PHF19 expression was positively associated with GBM progression, including cell proliferation, migration, invasion, chemosensitivity, and tumorigenesis. Using XAV-939, a Wnt/β-catenin inhibitor, we found that the effects of PHF19 on GBM cells were β-catenin-dependent. We also demonstrated that PHF19 expression was positively correlated with cytoplasmic β-catenin expression. PHF19 stabilized β-catenin by inhibiting the transcription of seven in absentia homolog 1 (SIAH1), an E3 ubiquitin ligase of β-catenin, through direct 1234567890():,; 1234567890():,; 1234567890():,; 1234567890():,; binding to the SIAH1 promoter region. Taken together, -
Identification and Preliminary Functional Analysis of Alternative Splicing of Siah1 in Xenopus Laevis
Biochemical and Biophysical Research Communications 396 (2010) 419–424 Contents lists available at ScienceDirect Biochemical and Biophysical Research Communications journal homepage: www.elsevier.com/locate/ybbrc Identification and preliminary functional analysis of alternative splicing of Siah1 in Xenopus laevis Luan Wen a,b, Jiantao Liu a,c, Yonglong Chen a, Donghai Wu a,* a Key Laboratory of Regenerative Biology, Guangzhou Institute of Biomedicine and Health, Chinese Academy of Sciences, 190 Kaiyuan Road, Guangzhou Science Park, 510530 Guangzhou, China b Graduate University of Chinese Academy of Sciences, 100049 Beijing, China c Laboratory of Veterinary Pharmacology, College of Veterinary Medicine South China Agricultural University, Guangzhou, China article info abstract Article history: Siah proteins are vertebrate homologs of the Drosophila ‘seven in absentia’ gene. In this study, we char- Received 11 April 2010 acterized two splicing forms, Siah1a and Siah1b, of the Xenopus seven in absentia homolog 1 gene (Siah1). Available online 22 April 2010 Overexpression of xSiah1a led to severe suppression of embryo cleavage, while that of xSiah1b was not effective even at a high dose. Competition analysis demonstrated that co-expression of xSiah1a and 1b Keywords: generated the same phenotype as overexpression of xSiah1a alone, suggesting that xSiah1b does not Xenopus interfere with the function of xSiah1a. Since xSiah1b has an additional 31 amino acids in the N-terminus xSiah1a compared to xSiah1a, progressive truncation of xSiah1b from the N-terminus showed that inability of xSiah1b xSiah1b to affect embryo cleavage was associated with the length of the N-terminal extension of extra amino acids. The possible implication of this finding is discussed. -
The SIAH1-HIPK2-P53ser46 Damage Response Pathway Is Involved in Temozolomide- Induced Glioblastoma Cell Death
Author Manuscript Published OnlineFirst on February 22, 2019; DOI: 10.1158/1541-7786.MCR-18-1306 Author manuscripts have been peer reviewed and accepted for publication but have not yet been edited. The SIAH1-HIPK2-p53ser46 damage response pathway is involved in temozolomide- induced glioblastoma cell death Yang He, Wynand P. Roos, Qianchao Wu, Thomas G. Hofmann and Bernd Kaina* Institute of Toxicology, University Medical Center Mainz, Obere Zahlbacher Str. 67, D-55131 Mainz, Germany *Corresponding author Bernd Kaina Department of Toxicology University Medical Center Mainz Obere Zahlbacher Str. 67 D-55131 Mainz, Germany Tel. 0049-6131-17-9217 Fax. 0049-6131-17-8499 E-mail: [email protected] Running title: HIPK2-SIAH1 in p53 mediated glioblastoma cell death Conflict of interest statement: Authors declare there is no conflict of interest. Key words: apoptosis, cell death, HIPK2, SIAH1, p53, DNA damage response, DNA repair, MGMT, glioblastoma, temozolomide, cancer therapy 1 Downloaded from mcr.aacrjournals.org on October 1, 2021. © 2019 American Association for Cancer Research. Author Manuscript Published OnlineFirst on February 22, 2019; DOI: 10.1158/1541-7786.MCR-18-1306 Author manuscripts have been peer reviewed and accepted for publication but have not yet been edited. Abstract Patients suffering from glioblastoma have a dismal prognosis, indicating the need for new therapeutic targets. Here we provide evidence that the DNA damage kinase HIPK2 and its negative regulatory E3-ubiquitin ligase SIAH1 are critical factors controlling temozolomide-induced cell death. We show that HIPK2 downregulation (HIPK2kd) significantly reduces the level of apoptosis. This was not the case in glioblastoma cells expressing the repair protein MGMT, suggesting that the primary DNA lesion responsible for triggering HIPK2-mediated apoptosis is O6-methylguanine.