Recent Insights Into the Mechanism of Glucocorticosteroid- Induced Apoptosis

Total Page:16

File Type:pdf, Size:1020Kb

Recent Insights Into the Mechanism of Glucocorticosteroid- Induced Apoptosis Cell Death and Differentiation (2002) 9, 6±19 ã 2002 Nature Publishing Group All rights reserved 1350-9047/02 $25.00 www.nature.com/cdd Review Recent insights into the mechanism of glucocorticosteroid- induced apoptosis CW Distelhorst*,1 apoptosis.1 But progress in understanding corticosteroid- induced apoptosis has lagged behind remarkable advances in 1 Division of Hematology/Oncology and Comprehensive Cancer Center, understanding other forms of apoptosis, such as that induced Departments of Medicine and Pharmacology, Case Western Reserve by the ligands of death receptors (e.g., Fas/CD95, tumor University School of Medicine and University Hospitals of Cleveland, necrosis factor). In part, this is because corticosteroid-induced Cleveland, Ohio, USA apoptosis is initiated by a steroid receptor-mediated change in * Corresponding author: CW Distelhorst, Division of Hematology/Oncology, Case Western Reserve University, 10900 Euclid Avenue, Cleveland, Ohio, gene expression, but specific genes that mediate cell death in OH 44106-4937, USA. Tel: 216-368-1175; Fax: 216-368-1166; response to corticosteroid treatment have not been identified. E-mail: [email protected] Recent findings have shed light on events in the cell death pathway downstream from gene regulation, including the Received 20.7.01; revised 7.9.01; accepted 3.10.01 caspases responsible for the execution phase of cell death, Edited by G Nunez the unexpected involvement of the multicatalytic proteasome in the death process, the suppression of prosurvival transcrip- tion factors (e.g., AP-1, c-myc,NF-kB), and crosstalk between Abstract T cell receptor and cytokine signaling pathways. Moreover, Glucocorticosteroid hormones induce apoptosis in lympho- evidence that mitochondrial dysfunction lies downstream of cytes. Therefore, glucocorticoids are commonly used as caspase activation in corticosteroid-induced apoptosis chal- immunosuppressive and chemotherapeutic agents. This lenges the generalization that mitochondrial dysfunction is the review examines many facets of the process by which central decision point in all forms of apoptosis. glucocorticoids induce apoptosis. This process is divided This review focuses primarily on insights gained in recent years regarding the mechanism of corticosteroid-induced into three stages, an initiation stage that involves glucocorti- apoptosis. Ultimately, the goal of investigating this form of coid receptor-mediated gene regulation, a decision stage that apoptosis is to better understand the role of corticosteroid involves the counterbalancing influence of prosurvival and hormones in physiological regulation of the immune system proapoptotic factors, and the execution stage which involves and in the treatment of lymphoid leukemias and lymphomas. caspase and endonuclease activation. Many aspects of To organize this review, corticosteroid-induced apoptosis glucocorticoid-induced apoptosis, such as mitochondrial is arbitrarily divided into three stages: the initiation stage, the dysfunction and caspase activation, are important steps in decision stage, and the execution stage (Figure 1). The virtually all forms of apoptosis. But the process glucocorti- initiation stage involves corticosteroid receptor-mediated coid-induced apoptosis differs from other forms of apoptosis changes in gene expression, while the execution stage in terms of initiation at the transcriptional level and involves activation of caspases, or other proteases and involvement of the multicatalytic proteasome and calcium. endonucleases. The decision stage includes those events that ultimately result in a commitment to cell death. A decision Moreover, the abundant opportunity for crosstalk between the to die appears to be made by counterbalancing actions of glucocorticoid receptor and other signaling pathways cytokines, which induce transcription factors that promote cell increases the complexity of glucocorticoid-induced apopto- proliferation and survival, versus the actions of corticoster- sis and its regulation. oids, which oppose the expression of these transcription Cell Death and Differentiation (2002) 9, 6 ± 19. DOI: 10.1038/sj/ factors. An important feature of corticosteroid-induced cdd/4400969 apoptosis is involvement of the multicatalytic proteasome, which appears to play an active role in promoting cell death by Keywords: glucocorticosteroid; dexamethasone; apoptosis; leu- degrading factors necessary for cell survival. kemia; lymphoma Abbreviations: IL, interleukin; AP-1, activating protein-1; CTLL, The initiation stage cytotoxic T lymphocytic leukemia; NFAT, nuclear factor of activated Glucocorticoid receptor mediated changes in gene T lymphocytes transcription In this review, the initiation stage is defined as the sequence of events leading up to and including direct regulation of gene Introduction transcription by the corticosteroid receptor (Figure 2). But The induction of apoptosis in thymocytes by glucocorticoster- what genes are regulated so that the lymphoid cell ultimately oid hormones is one of the earliest recognized forms of dies? The answer to this question has evaded investigators Glucocorticoid-induced apoptosis CW Distelhorst 7 Figure 1 Stages of glucocorticoid-induced apoptosis. Between the Initiation Stage and the Execution Stage lies the Decision Stage in which counterbalancing factors determine the commitment of the cell to live or die. Recent findings indicate that repression of prosurvival transcription factors and proteasome-mediated degradation of apoptosis inhibitory factors may play a crucial role in this stage genes involved in cell growth, differentiation and transforma- tion,3,4 their findings suggested that interference with transcription factors required for cell survival, such as AP-1 (activating protein-1), may contribute to apoptosis induction.2 This interpretation is consistent with earlier findings indicating that in cytokine (IL-2)-dependent CTLL cells there is a correlation between the presence of AP-1 DNA binding activity and repression of corticosteroid-induced apoptosis.5 In the absence of interleukin-2, corticosteroid treatment stimulates AP-1 degradation and induces apoptosis.5 The repression of AP-1 transcription factor activity in Figure 2 The Initiation Stage. There is considerable evidence that corticosteroid-treated lymphocytes is accompanied by glucocorticoid-induced apoptosis is initiated by the transactivation activity of the glucocorticoid receptor, but the specific genes regulated in this process repression of other prosurvival transcription factors, includ- 6 are not yet identified. RAP46/BAG1 represses the transactivation activity of ing the nuclear factor of activated T lymphocytes (NFAT). the glucocorticoid receptor, and therefore inhibits glucocorticoid-induced NFAT is composed of two components, NFATp and AP-1; apoptosis. Glucocorticoid-induced apoptosis is also repressed by T cell the dexamethasone-induced decrease in NFAT activity is receptor (TCR) signaling, which appears to alter the transactivation activity of due to a decrease in AP-1 activity.6 Also, Thompson and the glucocorticoid receptor. On the other hand, DAP3 increases glucocorti- 7 coid-induced apoptosis by enhancing the transactivation activity of the coworkers have provided substantial evidence that down- glucocorticoid receptor regulation of c-Myc accompanies the induction of apoptosis by corticosteroids, and that c-Myc overexpression inhibits corticosteroid-induced apoptosis. up to the present time. In part, the difficulty has been knowing what to look for: the induction of a gene or set of genes that produce cell death as a direct or indirect consequence of their Evidence for involvement of the glucocorticoid biological action, or the repression of genes necessary for receptor transactivation activity in maintenance of cell survival, or a combination of both? One glucocorticoid-induced apoptosis specific point of confusion in understanding corticosteroid- Recent evidence from several laboratories indicates that the induced apoptosis has to do with whether the transactivation transactivation activity of the glucocorticoid receptor is or transrepression functions of the corticosteroid receptor required to mediate corticosteroid-induced apoptosis. Chap- initiate the process of cell death. Evidence in favor of each is man et al 8 found that chimeric receptors containing the potent summarized here. VP16 and E1A viral transactivation domains in place of the corticosteroid receptor amino terminus enhanced the sensi- tivity of lymphoid cells to corticosteroid-induced apoptosis, Evidence for involvement of the glucocorticoid supporting a role for transactivation in apoptosis induction. receptor transrepression activity in Ramdas et al 9 addressed the question by comparing human glucocorticoid-induced apoptosis leukemic T cells that express either wild type corticosteroid The concept that corticosteroid-induced apoptosis is receptor or receptor that has a mutation that is activation mediated by the transrepression activity of the glucocorticoid deficient, but that retains the ability to repress AP-1 activity. receptor was largely based on the work of Helmberg et al.2 This mutant failed to mediate corticosteroid-induced apopto- These investigators reported that mutant glucocorticoid sis, indicating that the transactivation function of the receptor receptors defective in transactivation function, but fully is essential for corticosteroid-induced apoptosis. In a similar competent at interfering with AP-1 (activating protein-1) approach in vivo,
Recommended publications
  • Role of Mitochondrial Ribosomal Protein S18-2 in Cancerogenesis and in Regulation of Stemness and Differentiation
    From THE DEPARTMENT OF MICROBIOLOGY TUMOR AND CELL BIOLOGY (MTC) Karolinska Institutet, Stockholm, Sweden ROLE OF MITOCHONDRIAL RIBOSOMAL PROTEIN S18-2 IN CANCEROGENESIS AND IN REGULATION OF STEMNESS AND DIFFERENTIATION Muhammad Mushtaq Stockholm 2017 All previously published papers were reproduced with permission from the publisher. Published by Karolinska Institutet. Printed by E-Print AB 2017 © Muhammad Mushtaq, 2017 ISBN 978-91-7676-697-2 Role of Mitochondrial Ribosomal Protein S18-2 in Cancerogenesis and in Regulation of Stemness and Differentiation THESIS FOR DOCTORAL DEGREE (Ph.D.) By Muhammad Mushtaq Principal Supervisor: Faculty Opponent: Associate Professor Elena Kashuba Professor Pramod Kumar Srivastava Karolinska Institutet University of Connecticut Department of Microbiology Tumor and Cell Center for Immunotherapy of Cancer and Biology (MTC) Infectious Diseases Co-supervisor(s): Examination Board: Professor Sonia Lain Professor Ola Söderberg Karolinska Institutet Uppsala University Department of Microbiology Tumor and Cell Department of Immunology, Genetics and Biology (MTC) Pathology (IGP) Professor George Klein Professor Boris Zhivotovsky Karolinska Institutet Karolinska Institutet Department of Microbiology Tumor and Cell Institute of Environmental Medicine (IMM) Biology (MTC) Professor Lars-Gunnar Larsson Karolinska Institutet Department of Microbiology Tumor and Cell Biology (MTC) Dedicated to my parents ABSTRACT Mitochondria carry their own ribosomes (mitoribosomes) for the translation of mRNA encoded by mitochondrial DNA. The architecture of mitoribosomes is mainly composed of mitochondrial ribosomal proteins (MRPs), which are encoded by nuclear genomic DNA. Emerging experimental evidences reveal that several MRPs are multifunctional and they exhibit important extra-mitochondrial functions, such as involvement in apoptosis, protein biosynthesis and signal transduction. Dysregulations of the MRPs are associated with severe pathological conditions, including cancer.
    [Show full text]
  • IPS-1 Is Crucial for DAP3-Mediated Anoikis Induction by Caspase-8 Activation
    Cell Death and Differentiation (2009) 16, 1615–1621 & 2009 Macmillan Publishers Limited All rights reserved 1350-9047/09 $32.00 www.nature.com/cdd IPS-1 is crucial for DAP3-mediated anoikis induction by caspase-8 activation H-M Li1, D Fujikura1, T Harada1, J Uehara1, T Kawai2,3, S Akira2,3, JC Reed4, A Iwai1 and T Miyazaki*,1 Detachment of adherent epithelial cells from the extracellular matrix induces apoptosis, a process known as anoikis. We have shown that DAP3 is critical for anoikis induction. However, the mechanism for anoikis induction mediated by DAP3 is still unclear. Here, we show that interferon-b promoter stimulator 1 (IPS-1) binds DAP3 and induces anoikis by caspase activation. Recently, IPS-1 has been shown to be critical for antiviral immune responses, although there has been no report of its function in apoptosis induction. We show that overexpression of IPS-1 induces apoptosis by activation of caspase-3, -8, and -9. In addition, IPS-1 knockout mouse embryonic fibroblasts were shown to be resistant to anoikis. Interestingly, IPS-1 expression, recruitment of caspase-8 to IPS-1, and caspase-8 activation were induced after cell detachment. Furthermore, DAP3-mediated anoikis induction was inhibited by knockdown of IPS-1 expression. Therefore, we elucidated a novel function of IPS-1 for anoikis induction by caspase-8 activation. Cell Death and Differentiation (2009) 16, 1615–1621; doi:10.1038/cdd.2009.97; published online 31 July 2009 Apoptosis, the programmed physiological cell death, is critical induces programmed cell death, a phenomenon known as for modeling tissues and maintaining homeostasis in multi- anoikis.
    [Show full text]
  • Learning Protein Constitutive Motifs from Sequence Data Je´ Roˆ Me Tubiana, Simona Cocco, Re´ Mi Monasson*
    TOOLS AND RESOURCES Learning protein constitutive motifs from sequence data Je´ roˆ me Tubiana, Simona Cocco, Re´ mi Monasson* Laboratory of Physics of the Ecole Normale Supe´rieure, CNRS UMR 8023 & PSL Research, Paris, France Abstract Statistical analysis of evolutionary-related protein sequences provides information about their structure, function, and history. We show that Restricted Boltzmann Machines (RBM), designed to learn complex high-dimensional data and their statistical features, can efficiently model protein families from sequence information. We here apply RBM to 20 protein families, and present detailed results for two short protein domains (Kunitz and WW), one long chaperone protein (Hsp70), and synthetic lattice proteins for benchmarking. The features inferred by the RBM are biologically interpretable: they are related to structure (residue-residue tertiary contacts, extended secondary motifs (a-helixes and b-sheets) and intrinsically disordered regions), to function (activity and ligand specificity), or to phylogenetic identity. In addition, we use RBM to design new protein sequences with putative properties by composing and ’turning up’ or ’turning down’ the different modes at will. Our work therefore shows that RBM are versatile and practical tools that can be used to unveil and exploit the genotype–phenotype relationship for protein families. DOI: https://doi.org/10.7554/eLife.39397.001 Introduction In recent years, the sequencing of many organisms’ genomes has led to the collection of a huge number of protein sequences, which are catalogued in databases such as UniProt or PFAM Finn et al., 2014). Sequences that share a common ancestral origin, defining a family (Figure 1A), *For correspondence: are likely to code for proteins with similar functions and structures, providing a unique window into [email protected] the relationship between genotype (sequence content) and phenotype (biological features).
    [Show full text]
  • Co-Chaperone Potentiation of Vitamin D Receptor-Mediated Transactivation
    81 Co-chaperone potentiation of vitamin D receptor-mediated transactivation: a role for Bcl2-associated athanogene-1 as an intracellular-binding protein for 1,25-dihydroxyvitamin D3 R F Chun, M Gacad, L Nguyen, M Hewison and J S Adams Division of Endocrinology, Diabetes and Metabolism, Burns and Allen Research Institute, Cedars-Sinai Medical Center, Room D-3088, 8700 Beverly Boulevard, Los Angeles, California 90048, USA (Requests for offprints should be addressed to M Hewison; Email: [email protected]) Abstract The constitutively expressed member of the heat shock protein-70 family (hsc70) is a chaperone with multiple functions in cellular homeostasis. Previously, we demonstrated the ability of hsc70 to bind 25-hydroxyvitamin D3 (25-OHD3) and 1,25- dihydroxyvitamin D3 (1,25(OH)2D3). Hsc70 also recruits and interacts with the co-chaperone Bcl2-associated athanogene (BAG)-1 via the ATP-binding domain that resides on hsc70. Competitive ligand-binding assays showed that, like hsc70, recombinant BAG-1 is able to bind 25-OHD3 (KdZ0.71G0.25 nM, BmaxZ69.9G16.1 fmoles/mg protein) and 1,25(OH)2D3 (KdZ0.16G0.07 nM, BmaxZ38.1G3.5 fmoles/mg protein; both nZ3 separate binding assays, P!0.001 for Kd and Bmax). To investigate the functional significance of this, we transiently overexpressed the S, M, and L variants of BAG-1 into human kidney HKC-8 cells stably transfected with a 1,25(OH)2D3-responsive 24-hydroxylase (CYP24) promoter–reporter construct. As HKC-8 cells also express the enzyme 1a-hydroxylase, both 25-OHD3 (200 nM) and 1,25(OH)2D3 (5 nM) were able to induce CYP24 promoter activity.
    [Show full text]
  • Hnoa1 Interacts with Complex I and DAP3 and Regulates
    Supplemental Material can be found at: http://www.jbc.org/cgi/content/full/M807797200/DC1 THE JOURNAL OF BIOLOGICAL CHEMISTRY VOL. 284, NO. 8, pp. 5414–5424, February 20, 2009 © 2009 by The American Society for Biochemistry and Molecular Biology, Inc. Printed in the U.S.A. hNOA1 Interacts with Complex I and DAP3 and Regulates Mitochondrial Respiration and Apoptosis*□S Received for publication, October 9, 2008, and in revised form, December 15, 2008 Published, JBC Papers in Press, December 22, 2008, DOI 10.1074/jbc.M807797200 Tingdong Tang‡, Bin Zheng‡1, Sheng-hong Chen‡§, Anne N. Murphy¶, Krystyna Kudlicka‡, Huilin Zhou‡§, and Marilyn G. Farquhar‡2 From the Departments of ‡Cellular and Molecular Medicine and ¶Pharmacology and §Ludwig Institute for Cancer Research, University of California, San Diego, La Jolla, California 92093-0651 Mitochondria are dynamic organelles that play key roles in fusion proteins such as the dynamin like large G proteins Drp1 metabolism, energy production, and apoptosis. Coordination of and Opa1, the cell’s susceptibility to apoptotic agents (3) or these processes is essential to maintain normal cellular func- ability to generate ATP (4, 5) is altered. tions. Here we characterized hNOA1, the human homologue of Apoptosis is controlled by a diverse range of cell signals, AtNOA1 (Arabidopsis thaliana nitric oxide-associated protein which may originate either extracellularly (extrinsic inducers) Downloaded from 1), a large mitochondrial GTPase. By immunofluorescence, or intracellularly (intrinsic inducers), and mitochondria play immunoelectron microscopy, and mitochondrial subfraction- central roles in both pathways (6). The apoptotic pathways ation, endogenous hNOA1 is localized within mitochondria involve a growing list of mitochondria-associated proteins, where it is peripherally associated with the inner mitochondrial such as Bad, cytochrome c, Smac, AIF, Bcl-2, and others, most membrane facing the mitochondrial matrix.
    [Show full text]
  • Insight Into Glucocorticoid Receptor Signalling Through Interactome Model Analysis
    RESEARCH ARTICLE Insight into glucocorticoid receptor signalling through interactome model analysis Emyr Bakker1, Kun Tian1, Luciano Mutti1, Constantinos Demonacos2, Jean- Marc Schwartz2☯*, Marija Krstic-Demonacos1☯* 1 Biomedical Research Centre, School of Environment and Life Sciences, University of Salford, Salford, United Kingdom, 2 Faculty of Biology, Medicine and Health, University of Manchester, Manchester, United Kingdom ☯ These authors contributed equally to this work. * [email protected] (JMS); [email protected] (MKD) a1111111111 a1111111111 a1111111111 a1111111111 Abstract a1111111111 Glucocorticoid hormones (GCs) are used to treat a variety of diseases because of their potent anti-inflammatory effect and their ability to induce apoptosis in lymphoid malignan- cies through the glucocorticoid receptor (GR). Despite ongoing research, high glucocorticoid efficacy and widespread usage in medicine, resistance, disease relapse and toxicity remain OPEN ACCESS factors that need addressing. Understanding the mechanisms of glucocorticoid signalling Citation: Bakker E, Tian K, Mutti L, Demonacos C, and how resistance may arise is highly important towards improving therapy. To gain insight Schwartz J-M, Krstic-Demonacos M (2017) Insight into this we undertook a systems biology approach, aiming to generate a Boolean model of into glucocorticoid receptor signalling through interactome model analysis. PLoS Comput Biol 13 the glucocorticoid receptor protein interaction network that encapsulates functional relation- (11): e1005825. https://doi.org/10.1371/journal. ships between the GR, its target genes or genes that target GR, and the interactions pcbi.1005825 between the genes that interact with the GR. This model named GEB052 consists of 52 Editor: Andrey Rzhetsky, University of Chicago, nodes representing genes or proteins, the model input (GC) and model outputs (cell death UNITED STATES and inflammation), connected by 241 logical interactions of activation or inhibition.
    [Show full text]
  • Bag6 Complex Contains a Minimal Tail-Anchor–Targeting Module and a Mock BAG Domain
    Bag6 complex contains a minimal tail-anchor–targeting module and a mock BAG domain Jee-Young Mocka, Justin William Chartrona,Ma’ayan Zaslavera,YueXub,YihongYeb, and William Melvon Clemons Jr.a,1 aDivision of Chemistry and Chemical Engineering, California Institute of Technology, Pasadena, CA 91125; and bLaboratory of Molecular Biology, National Institute of Diabetes and Digestive and Kidney Diseases, National Institutes of Health, Bethesda, MD 20892 Edited by Gregory A. Petsko, Weill Cornell Medical College, New York, NY, and approved December 1, 2014 (received for review February 12, 2014) BCL2-associated athanogene cochaperone 6 (Bag6) plays a central analogous yeast complex contains two proteins, Get4 and Get5/ role in cellular homeostasis in a diverse array of processes and is Mdy2, which are homologs of the mammalian proteins TRC35 part of the heterotrimeric Bag6 complex, which also includes and Ubl4A, respectively. In yeast, these two proteins form ubiquitin-like 4A (Ubl4A) and transmembrane domain recognition a heterotetramer that regulates the handoff of the TA protein complex 35 (TRC35). This complex recently has been shown to be from the cochaperone small, glutamine-rich, tetratricopeptide important in the TRC pathway, the mislocalized protein degrada- repeat protein 2 (Sgt2) [small glutamine-rich tetratricopeptide tion pathway, and the endoplasmic reticulum-associated degrada- repeat-containing protein (SGTA) in mammals] to the delivery tion pathway. Here we define the architecture of the Bag6 factor Get3 (TRC40 in mammals) (19–22). It is expected that the complex, demonstrating that both TRC35 and Ubl4A have distinct mammalian homologs, along with Bag6, play a similar role (23– C-terminal binding sites on Bag6 defining a minimal Bag6 complex.
    [Show full text]
  • Apiap2 Transcription Factor Restricts Development of the Toxoplasma Tissue Cyst
    ApiAP2 transcription factor restricts development of the Toxoplasma tissue cyst Joshua B. Radkea,1, Olivier Lucasa,1,2, Erandi K. De Silvab, YanFen Mac, William J. Sullivan, Jr.d, Louis M. Weissc, Manuel Llinasb, and Michael W. Whitea,3 aDepartments of Molecular Medicine and Global Health, University of South Florida, Tampa, FL 33612; bDepartment of Molecular Biology and Lewis-Sigler Institute for Integrative Genomics, Princeton University, Princeton, NJ 08544; cDepartments of Medicine and Microbiology and Immunology, Albert Einstein College of Medicine, Bronx, NY 10461; and dDepartment of Pharmacology and Toxicology, Indiana University School of Medicine, Indianapolis, IN 46202 Edited* by Jitender P. Dubey, US Department of Agriculture, Beltsville, MD, and approved March 14, 2013 (received for review January 3, 2013) Cellular differentiation leading to formation of the bradyzoite tissue the cell cycle transcriptome of Plasmodium falciparum and Toxo- cyst stage is the underlying cause of chronic toxoplasmosis. Con- plasma asexual stages are progressive with little understanding yet sequently, mechanisms responsible for controlling development in as to how these serial patterns are controlled (8, 9). In Toxoplasma, the Toxoplasma intermediate life cycle have long been sought. Here, conversion between developmental stages is also linked to signif- we identified 15 Toxoplasma mRNAs induced in early bradyzoite icant changes in gene expression (5). Data mining of genome development that encode proteins with apicomplexan AP2 (ApiAP2) sequences has recently identified a family of plant-related AP2 DNA binding domains. Of these 15 mRNAs, the AP2IX-9 mRNA dem- domain (DNA binding) containing proteins in the Apicomplexa onstrated the largest expression increase during alkaline-induced [apicomplexan AP2 (ApiAP2) proteins] (10).
    [Show full text]
  • Short Peptides Derived from the BAG-1 C-Terminus Inhibit the Interaction Between BAG-1 and HSC70 and Decrease Breast Cancer Cell Growth
    FEBS Letters 583 (2009) 3405–3411 journal homepage: www.FEBSLetters.org Short peptides derived from the BAG-1 C-terminus inhibit the interaction between BAG-1 and HSC70 and decrease breast cancer cell growth Adam Sharp a, Ramsey I. Cutress a, Peter W.M. Johnson a, Graham Packham a, Paul A. Townsend b,* a Cancer Research UK Centre, Cancer Sciences Division, University of Southampton, School of Medicine, Southampton General Hospital, Southampton S016 6YD, UK b Human Genetics Division, University of Southampton, School of Medicine, Southampton General Hospital, Southampton S016 6YD, UK article info abstract Article history: BAG-1, a multifunctional protein, interacts with a plethora of cellular targets where the interaction Received 8 July 2009 with HSC70 and HSP70, is considered vital. Structural studies have demonstrated the C-terminal of Revised 21 September 2009 BAG-1 forms a bundle of three alpha-helices of which helices 2 and 3 are directly involved in binding Accepted 24 September 2009 to the chaperones. Here we found peptides derived from helices 2 and 3 of BAG-1 interfered with Available online 1 October 2009 BAG-1:HSC70 binding. We confirmed that a 12 amino-acid peptide from helix 2 directly interacted Edited by Lukas Huber with HSC70 and when introduced into MCF-7 and ZR-75-1 cells, these peptides inhibited their growth. In conclusion, we have identified a small domain within BAG-1 which appears to play a crit- ical role in the interaction with HSC70. Keywords: BAG-1 HSC70 Structured summary: HSP70 MINT-7265269, MINT-7265296, MINT-7265324, MINT-7265339, MINT-7265351, MINT-7265364, MINT- Interaction 7265483, MINT-7265464, MINT-7265310: HSC70 (uniprotkb:P11142) binds (MI:0407) to BAG1 (uni- Binding protkb:Q99933) by peptide array (MI:0081) MINT-7265281: peptide 15L (uniprotkb:Q99933) binds (MI:0407) to HSC70 (uniprotkb:P11142) by surface plasmon resonance (MI:0107) Ó 2009 Federation of European Biochemical Societies.
    [Show full text]
  • A High-Throughput Approach to Uncover Novel Roles of APOBEC2, a Functional Orphan of the AID/APOBEC Family
    Rockefeller University Digital Commons @ RU Student Theses and Dissertations 2018 A High-Throughput Approach to Uncover Novel Roles of APOBEC2, a Functional Orphan of the AID/APOBEC Family Linda Molla Follow this and additional works at: https://digitalcommons.rockefeller.edu/ student_theses_and_dissertations Part of the Life Sciences Commons A HIGH-THROUGHPUT APPROACH TO UNCOVER NOVEL ROLES OF APOBEC2, A FUNCTIONAL ORPHAN OF THE AID/APOBEC FAMILY A Thesis Presented to the Faculty of The Rockefeller University in Partial Fulfillment of the Requirements for the degree of Doctor of Philosophy by Linda Molla June 2018 © Copyright by Linda Molla 2018 A HIGH-THROUGHPUT APPROACH TO UNCOVER NOVEL ROLES OF APOBEC2, A FUNCTIONAL ORPHAN OF THE AID/APOBEC FAMILY Linda Molla, Ph.D. The Rockefeller University 2018 APOBEC2 is a member of the AID/APOBEC cytidine deaminase family of proteins. Unlike most of AID/APOBEC, however, APOBEC2’s function remains elusive. Previous research has implicated APOBEC2 in diverse organisms and cellular processes such as muscle biology (in Mus musculus), regeneration (in Danio rerio), and development (in Xenopus laevis). APOBEC2 has also been implicated in cancer. However the enzymatic activity, substrate or physiological target(s) of APOBEC2 are unknown. For this thesis, I have combined Next Generation Sequencing (NGS) techniques with state-of-the-art molecular biology to determine the physiological targets of APOBEC2. Using a cell culture muscle differentiation system, and RNA sequencing (RNA-Seq) by polyA capture, I demonstrated that unlike the AID/APOBEC family member APOBEC1, APOBEC2 is not an RNA editor. Using the same system combined with enhanced Reduced Representation Bisulfite Sequencing (eRRBS) analyses I showed that, unlike the AID/APOBEC family member AID, APOBEC2 does not act as a 5-methyl-C deaminase.
    [Show full text]
  • DAP3 Antibody A
    C 0 2 - t DAP3 Antibody a e r o t S Orders: 877-616-CELL (2355) [email protected] Support: 877-678-TECH (8324) 2 7 Web: [email protected] 1 www.cellsignal.com 2 # 3 Trask Lane Danvers Massachusetts 01923 USA For Research Use Only. Not For Use In Diagnostic Procedures. Applications: Reactivity: Sensitivity: MW (kDa): Source: UniProt ID: Entrez-Gene Id: WB H Endogenous 46 Rabbit P51398 7818 Product Usage Information 5. Imataka, H. et al. (1997) EMBO J 16, 817-25. 6. Levy-Strumpf, N. et al. (1997) Mol Cell Biol 17, 1615-25. Application Dilution 7. Henis-Korenblit, S. et al. (2000) Mol Cell Biol 20, 496-506. Western Blotting 1:1000 Storage Supplied in 10 mM sodium HEPES (pH 7.5), 150 mM NaCl, 100 µg/ml BSA and 50% glycerol. Store at –20°C. Do not aliquot the antibody. Specificity / Sensitivity DAP3 Antibody detects endogenous levels of total DAP3 protein. The antibody does not cross-react with other DAP family members. Species Reactivity: Human Source / Purification Polyclonal antibodies are produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Lys58 of human DAP3. Antibodies are purified by protein A and peptide affinity chromatography. Background Death associated protein 1 (DAP1) is a 15 kDa protein that functions as a positive mediator of cell death initiated by interferon-gamma (1,2). The DAP1 protein is proline rich and possesses one SH3 binding motif, as well as several consensus protein kinase phosphorylation sites (1). The protein is localized in the cytoplasm, but the detailed mechanism of its proapoptotic function is unclear.
    [Show full text]
  • LKB1 Is Crucial for TRAIL-Mediated Apoptosis Induction in Osteosarcoma
    ANTICANCER RESEARCH 27: 761-768 (2007) LKB1 is Crucial for TRAIL-mediated Apoptosis Induction in Osteosarcoma SHINTARO TAKEDA1,2, ATSUSHI IWAI2,3, MITSUKO NAKASHIMA2, DAISUKE FUJIKURA2,3, SATOKO CHIBA2,3, HONG MEI LI3, JUN UEHARA2,3, SATOSHI KAWAGUCHI1, MITSUNORI KAYA1, SATOSHI NAGOYA1, TAKURO WADA1, JUNYING YUAN4, SYDONIA RAYTER5, ALAN ASHWORTH5, JOHN C. REED6, TOSHIHIKO YAMASHITA1, TOSHIMITSU UEDE2 and TADAAKI MIYAZAKI2,3 1Department of Orthopaedic Surgery, Sapporo Medical University School of Medicine, Sapporo; 2Division of Molecular Immunology, Institute for Genetic Medicine, Hokkaido University, Sapporo; 3Department of Bioresources, Hokkaido University Research Center for Zoonosis Control, Sapporo, 0010021, Japan; 4Department of Cell Biology, Harvard Medical School, Boston, Massachusetts, 02115, U.S.A.; 5Cancer Research UK Gene Function and Regulation Group, The Breakthrough Breast Cancer Research Centre, Institute of Cancer Research, London SW3 6JB, U.K.; 6Burnham Institute for Medical Research, La Jolla, California, 92037, U.S.A. Abstract. Background: Despite improvements in Development of chemotherapy and surgery has resulted in chemotherapy and surgery in the treatment of osteosarcoma, the considerable improvement of survival rates of satisfactory results are still difficult to achieve. New therapeutic osteosarcoma patients (1), the current chemotherapeutic modalities need to be developed for the improvement of these agents still have a lot of problems to overcome, such as treatments. TRAIL (TNF-related apoptosis inducing ligand) is serious side-effects or resistance to multiple chemo- known as a selective apoptosis inducer in most tumor cells, but therapeutic agents in 20-30% of the patients (2). New not in normal cells. Therefore, TRAIL is a good candidate therapeutic modalities for these patients are driven by target for the treatment of tumors.
    [Show full text]