DOCSLIB.ORG

Substrates Caspase Activity and Cleavage of Caspase-8 T Cells

Total Page：16

File Type：pdf, Size：1020Kb

Download full-text PDF Read full-text
Substrates Caspase Activity and Cleavage of Caspase-8 T Cells Effector CD4+ T Cells Generate Intermediate Caspase Activity and Cleavage of Caspase-8 Substrates This information is current as Ravi S. Misra, Dawn M. Jelley-Gibbs, Jennifer Q. Russell, of October 1, 2021. Gail Huston, Susan L. Swain and Ralph C. Budd J Immunol 2005; 174:3999-4009; ; doi: 10.4049/jimmunol.174.7.3999 http://www.jimmunol.org/content/174/7/3999 Downloaded from References This article cites 64 articles, 27 of which you can access for free at: http://www.jimmunol.org/content/174/7/3999.full#ref-list-1 http://www.jimmunol.org/ Why The JI? Submit online. • Rapid Reviews! 30 days* from submission to initial decision • No Triage! Every submission reviewed by practicing scientists • Fast Publication! 4 weeks from acceptance to publication *average by guest on October 1, 2021 Subscription Information about subscribing to The Journal of Immunology is online at: http://jimmunol.org/subscription Permissions Submit copyright permission requests at: http://www.aai.org/About/Publications/JI/copyright.html Email Alerts Receive free email-alerts when new articles cite this article. Sign up at: http://jimmunol.org/alerts The Journal of Immunology is published twice each month by The American Association of Immunologists, Inc., 1451 Rockville Pike, Suite 650, Rockville, MD 20852 Copyright © 2005 by The American Association of Immunologists All rights reserved. Print ISSN: 0022-1767 Online ISSN: 1550-6606. The Journal of Immunology Effector CD4؉ T Cells Generate Intermediate Caspase Activity and Cleavage of Caspase-8 Substrates1 Ravi S. Misra,* Dawn M. Jelley-Gibbs,† Jennifer Q. Russell,* Gail Huston,† Susan L. Swain,† and Ralph C. Budd2* Caspase-8 activation promotes cell apoptosis but is also essential for T cell activation. The extent of caspase activation and substrate cleavage in these divergent processes remains unclear. We show that murine effector CD4؉ T cells generated levels of caspase activity intermediate between unstimulated T cells and apoptotic populations. Both caspase-8 and caspase-3 were partially activated in effector T cells, which was reflected in cleavage of the caspase-8 substrates, c-FLIPL, receptor interacting protein 1, and to a lesser extent Bid, but not the caspase-3 substrate inhibitor of caspase-activated DNase. Th2 effector CD4؉ T cells manifested more caspase activity than did Th1 effectors, and caspase blockade greatly decreased initiation of cell cycling. The current findings define the level of caspase activity and substrates during initiation of T cell cycling. The Journal of Immunology, 2005, 174: 3999–4009. Downloaded from t is becoming increasingly evident that ligation of death re- The earlier view that caspase activation occurs only through its ceptors and activation of cellular caspases does not necessar- cleavage has been revised for certain upstream caspases. An alter- I ily always result in apoptosis. Involvement of the death re- native method of caspase activation is through a conformational ceptor, Fas, in processes other than cell death has been revealed by change. For example, when caspase-9 interacts with cytochrome c the ability of Fas ligation to enhance neural outgrowth (1), cardiac complexed to Apaf-1, a conformational change of caspase-9 ex- hypertrophy (2), liver regeneration (3), fibroblast proliferation (4), poses its active site (16, 17). In a similar manner, procaspase-8 can http://www.jimmunol.org/ dendritic cell maturation (5), and T cell activation (6). More di- induce a structural change in a neighboring procaspase-8 molecule, rectly, caspase-8 plays an integral role in T cell function, as hu- resulting in active full-length caspase-8 (10). In addition, the en- mans bearing nonfunctional caspase-8, as well as caspase-8 con- zymatically inactive casapase-8-like molecule c-FLIPL can also ditional knockout mice, exhibit severe defects in T cell activation associate with and activate full-length caspase-8 (18). As c-FLIPL (7, 8). However, there is currently no clear picture of which also contains a known caspase cleavage site, it represents poten- caspases or which substrates are involved with T cell activation tially one of the earliest substrates of caspase-8 (19). and how far down the caspase cascade these signals propagate. The significance of caspase activity in nonapoptotic effector T Traditionally, activation of the upstream initiator caspases that cells is still poorly understood. It is not known how the levels of by guest on October 1, 2021 possess large prodomains, such as procaspases-8, -9, and -10, had active caspases or cleavage of their substrates in viable effector T been thought to require proteolytic cleavage of the zymogen, resulting cells compare with those in resting T cells or those undergoing in the release of large and small subunits (9). These fragments then apoptosis after Fas ligation. We observe that intermediate levels of heterodimerize to form an active caspase molecule. Typically, the caspase activity are generated at the effector stage of T cell acti- cleavage occurs by auto- and cross-proteolysis in response to the li- vation, which are proportional to cell cycling, and that blockade of gation of death receptors, which increases the local concentrations of caspase activity inhibits cell cycling. Proximal caspase-8 sub- caspases (9–11). The upstream initiator caspase signal is propagated strates such as c-FLIPL and receptor interacting protein 1 (RIP1) to the effector procaspases, such as procaspase-3, by proteolytic cleav- are cleaved in effector T cells, but not the downstream caspase-3 age of the procaspase. As part of this caspase cascade, various caspase substrate ICAD. Furthermore, the caspase activity and cleavage of substrates are cleaved, including the DNase inhibitor inhibitor of substrates is more extensive in Th2 effector cells than in Th1 ef- caspase-activated DNase (ICAD)3 (12) and structural proteins, such fectors. Collectively, these results define the caspase activity pro- as actin (13), fodrin (14), and lamin (15). file in viable T cells and underscore the functional requirements of such activity. *University of Vermont, Immunobiology Program, College of Medicine, Burlington, Materials and Methods VT 05405; and †Trudeau Institute, Saranac Lake, NY 12983 Mice Received for publication July 14, 2004. Accepted for publication January 11, 2005. C57BL/6 mice were housed and bred in the University of Vermont animal The costs of publication of this article were defrayed in part by the payment of page facility and were used at 2–6 mo of age. B10.Br V␣11/V␤3 AND TCR charges. This article must therefore be hereby marked advertisement in accordance transgenic (pigeon cytochrome c/I-Ek responsive) mice were used at 2–3 with 18 U.S.C. Section 1734 solely to indicate this fact. mo of age and were bred in the animal facilities at the Trudeau Institute. 1 This work was supported by grants from the National Institutes of Health (AI36333 Both facilities are American Association of Laboratory Animal Care ap- and AI45666 to R.C.B.; AI26887 and AI46530 to S.L.S.). R.S.M. is supported by proved and protocols were approved by the Institutional Animal Care and National Institutes of Health Grant No. 5T32ES007122. Use Committee at the respective institutions. 2 Address correspondence and reprint requests to Dr. Ralph C. Budd, University of Vermont, Immunobiology Program, College of Medicine, 89 Beaumont Avenue, T cell purification Given Building D305, Burlington, VT 05405. E-mail address: [email protected] 3 Abbreviations used in this paper: ICAD, inhibitor of caspase-activated DNase; C57BL/6 spleens and lymph nodes were isolated and disrupted through RIP1, receptor interacting protein 1; PCCF, pigeon cytochrome c fragment; z-VAD, nylon mesh in RPMI 1640 (MediaTech) containing 5% (v/v) FCS (Hy- z-Val-Ala-Asp-fluoromethylketone; FasL, Fas ligand; WCL, whole cell lysate; XIAP, Clone). Erythrocyte lysis of splenocytes was performed using Gey’s solu- ϩ X-linked inhibitor of apoptosis protein. tion. Lymphocytes and splenocytes were combined and CD4 T cells were Copyright © 2005 by The American Association of Immunologists, Inc. 0022-1767/05/$02.00 4000 EFFECTOR CD4 T CELLS GENERATE MODERATE CASPASE LEVELS isolated by negative selection. C57BL/6 cells were incubated with anti- Immunoblot analysis CD8 (Tib 105), anti-MHC class II (M5/114/15/2; a kind gift of M. Rinco´n, University of Vermont, Burlington, VT), anti-CD11b (M1/70), anti-NK1.1 Viable cells were lysed in buffer containing 0.2% Nonidet P-40, 20 mM (PK136), and anti-B220 (RA3-6B2) on ice for 45 min. For total T cell Tris-HCl (pH 7.4), 2 mM sodium orthovanadate, 10% glycerol, 150 mM ␮ isolation, anti-CD8 was omitted from Ab mix. Cells were washed three NaCl, complete protease inhibitor (Roche Diagnostics), and 10 M z-VAD times and rocked with goat anti-mouse and goat anti-rat conjugated mag- (Enzyme System Products). Protein concentration was determined by netic beads at a 10:1 ratio of beads to cell (Qiagen) at 4°C for 45 min. Bradford assay (Bio-Rad). Protein lysates were boiled for 5 min in loading Magnetic depletion was used to remove bead-bound cells, routinely yield- buffer containing 2-ME and were separated using SDS-PAGE on 12.5% ing Ͼ90% CD4ϩ T cells. Cells were washed and resuspended in culture gels. Proteins were transferred onto a polyvinylidene difluoride membrane medium (RPMI 1640 supplemented with 25 mM HEPES, 2.5 mg/ml glu- (Bio-Rad) and blocked using 4% milk in TBS plus 0.1% Tween 20 at room cose (Sigma-Aldrich), 10 ␮g/ml folate (Invitrogen Life Technologies), 110 temperature for 1 h. Membranes were incubated at 4°C overnight in milk Ϫ ␮ ␮g/ml pyruvate (Invitrogen Life Technologies), 5 ϫ 10 5 M 2-ME (Sig- containing one of the following primary detection Abs: 1 g/ml actin (I-19; ma-Aldrich), 292.3 ␮g/ml glutamine (Invitrogen Life Technologies), 100 Santa Cruz Biotechnology), Bid (R&D Systems), caspase-3 (a kind gift of U/ml penicillin-streptomycin (Invitrogen Life Technologies), and Y.
Load more

Recommended publications
  • Apoptotic Threshold Is Lowered by P53 Transactivation of Caspase-6
    Apoptotic threshold is lowered by p53 transactivation of caspase-6 Timothy K. MacLachlan*† and Wafik S. El-Deiry*‡§¶ʈ** *Laboratory of Molecular Oncology and Cell Cycle Regulation, Howard Hughes Medical Institute, and Departments of ‡Medicine, §Genetics, ¶Pharmacology, and ࿣Cancer Center, University of Pennsylvania School of Medicine, Philadelphia, PA 19104 Communicated by Britton Chance, University of Pennsylvania School of Medicine, Philadelphia, PA, April 23, 2002 (received for review January 11, 2002) Little is known about how a cell’s apoptotic threshold is controlled Inhibition of the enzyme reduces the sensitivity conferred by after exposure to chemotherapy, although the p53 tumor suppres- overexpression of p53. These results identify a pathway by which sor has been implicated. We identified executioner caspase-6 as a p53 is able to accelerate the apoptosis cascade by loading the cell transcriptional target of p53. The mechanism involves DNA binding with cell death proteases so that when an apoptotic signal is by p53 to the third intron of the caspase-6 gene and transactiva- received, programmed cell death occurs rapidly. tion. A p53-dependent increase in procaspase-6 protein level al- lows for an increase in caspase-6 activity and caspase-6-specific Materials and Methods Lamin A cleavage in response to Adriamycin exposure. Specific Western Blotting and Antibodies. Immunoblotting was carried out by inhibition of caspase-6 blocks cell death in a manner that correlates using mouse anti-human p53 monoclonal (PAb1801; Oncogene), with caspase-6 mRNA induction by p53 and enhances long-term rabbit anti-human caspase-3 (Cell Signaling, Beverly, MA), mouse survival in response to a p53-mediated apoptotic signal.
    [Show full text]
  • XIAP's Profile in Human Cancer
    biomolecules Review XIAP’s Profile in Human Cancer Huailu Tu and Max Costa * Department of Environmental Medicine, Grossman School of Medicine, New York University, New York, NY 10010, USA; [email protected] * Correspondence: [email protected] Received: 16 September 2020; Accepted: 25 October 2020; Published: 29 October 2020 Abstract: XIAP, the X-linked inhibitor of apoptosis protein, regulates cell death signaling pathways through binding and inhibiting caspases. Mounting experimental research associated with XIAP has shown it to be a master regulator of cell death not only in apoptosis, but also in autophagy and necroptosis. As a vital decider on cell survival, XIAP is involved in the regulation of cancer initiation, promotion and progression. XIAP up-regulation occurs in many human diseases, resulting in a series of undesired effects such as raising the cellular tolerance to genetic lesions, inflammation and cytotoxicity. Hence, anti-tumor drugs targeting XIAP have become an important focus for cancer therapy research. RNA–XIAP interaction is a focus, which has enriched the general profile of XIAP regulation in human cancer. In this review, the basic functions of XIAP, its regulatory role in cancer, anti-XIAP drugs and recent findings about RNA–XIAP interactions are discussed. Keywords: XIAP; apoptosis; cancer; therapeutics; non-coding RNA 1. Introduction X-linked inhibitor of apoptosis protein (XIAP), also known as inhibitor of apoptosis protein 3 (IAP3), baculoviral IAP repeat-containing protein 4 (BIRC4), and human IAPs like protein (hILP), belongs to IAP family which was discovered in insect baculovirus [1]. Eight different IAPs have been isolated from human tissues: NAIP (BIRC1), BIRC2 (cIAP1), BIRC3 (cIAP2), XIAP (BIRC4), BIRC5 (survivin), BIRC6 (apollon), BIRC7 (livin) and BIRC8 [2].
    [Show full text]
  • The Role of Caspase-2 in Regulating Cell Fate
    cells Review The Role of Caspase-2 in Regulating Cell Fate Vasanthy Vigneswara and Zubair Ahmed * Neuroscience and Ophthalmology, Institute of Inflammation and Ageing, University of Birmingham, Birmingham B15 2TT, UK; [email protected] * Correspondence: [email protected] Received: 15 April 2020; Accepted: 12 May 2020; Published: 19 May 2020 Abstract: Caspase-2 is the most evolutionarily conserved member of the mammalian caspase family and has been implicated in both apoptotic and non-apoptotic signaling pathways, including tumor suppression, cell cycle regulation, and DNA repair. A myriad of signaling molecules is associated with the tight regulation of caspase-2 to mediate multiple cellular processes far beyond apoptotic cell death. This review provides a comprehensive overview of the literature pertaining to possible sophisticated molecular mechanisms underlying the multifaceted process of caspase-2 activation and to highlight its interplay between factors that promote or suppress apoptosis in a complicated regulatory network that determines the fate of a cell from its birth and throughout its life. Keywords: caspase-2; procaspase; apoptosis; splice variants; activation; intrinsic; extrinsic; neurons 1. Introduction Apoptosis, or programmed cell death (PCD), plays a pivotal role during embryonic development through to adulthood in multi-cellular organisms to eliminate excessive and potentially compromised cells under physiological conditions to maintain cellular homeostasis [1]. However, dysregulation of the apoptotic signaling pathway is implicated in a variety of pathological conditions. For example, excessive apoptosis can lead to neurodegenerative diseases such as Alzheimer’s and Parkinson’s disease, whilst insufficient apoptosis results in cancer and autoimmune disorders [2,3]. Apoptosis is mediated by two well-known classical signaling pathways, namely the extrinsic or death receptor-dependent pathway and the intrinsic or mitochondria-dependent pathway.
    [Show full text]
  • Proteases to Die For
    Downloaded from genesdev.cshlp.org on September 27, 2021 - Published by Cold Spring Harbor Laboratory Press REVIEW Proteases to die for Vincent Cryns1 and Junying Yuan2,3 1Center for Endocrinology, Metabolism and Molecular Medicine, Northwestern University School of Medicine, Chicago, Illinois 60611 USA; 2Department of Cell Biology, Harvard Medical School, Boston, Massachusetts 02115 USA Apoptosis or programmed cell death (PCD) is a geneti- have identified two genes (ced-3 and ced-4) that are each cally regulated, cellular suicide mechanism that plays a required for the execution of cell death and one (ced-9) crucial role in development and in the defense of homeo- that inhibits cell death (Hengartner et al. 1992; Yuan and stasis. Cells respond to a variety of disparate signals by Horvitz 1992; Yuan et al. 1993). Mutational analyses of committing suicide through a series of dramatic but re- these genes in C. elegans have defined a sequential cell markably uniform events. Morphologically, cells under- death pathway. Inactivating mutations of ced-9 result in going apoptosis demonstrate nuclear/cytoplasmic con- inappropriate cell deaths, but only if both ced-3 and densation and membrane protrusions. These initial ced-4 are functional (Hengartner et al. 1992). Targeted changes are followed by fragmentation of the nuclear overexpression of either ced-4 or ced-3 induces cell contents and subsequent encapsulation of these frag- death, these cell deaths are inhibited by ced-9. In trans- ments into ‘‘apoptotic bodies’’ that are quickly and un- genic worms, maximal cell death induced by ced-4 over- obtrusively consumed by adjacent cells, thereby leaving expression requires ced-3, whereas ced-3-mediated cell little trace of the apoptotic cell’s prior existence (Kerr et death is independent of ced-4.
    [Show full text]
  • Bax Transmembrane Domain Interacts with Prosurvival Bcl-2 Proteins in Biological Membranes
    Bax transmembrane domain interacts with prosurvival Bcl-2 proteins in biological membranes Vicente Andreu-Fernándeza,b,c, Mónica Sanchoa, Ainhoa Genovésa, Estefanía Lucendoa, Franziska Todtb, Joachim Lauterwasserb,d, Kathrin Funkb,d, Günther Jahreise, Enrique Pérez-Payáa,f,1, Ismael Mingarroc,2, Frank Edlichb,g,2, and Mar Orzáeza,2 aLaboratory of Peptide and Protein Chemistry, Centro de Investigación Príncipe Felipe, E-46012 Valencia, Spain; bInstitute for Biochemistry and Molecular Biology, University of Freiburg, 79104 Freiburg, Germany; cDepartament de Bioquímica i Biologia Molecular, Estructura de Recerca Interdisciplinar en Biotecnología i Biomedicina, Universitat de València, 46100 Burjassot, Spain; dFaculty of Biology, University of Freiburg, 79104 Freiburg, Germany; eDepartment of Biochemistry/Biotechnology, Martin Luther University Halle-Wittenberg, 06120 Halle, Germany; fInstituto de Biomedicina de Valencia, Instituto de Biomedicina de Valencia–Consejo Superior de Investigaciones Científicas, 46010 Valencia, Spain; and gBIOSS, Centre for Biological Signaling Studies, University of Freiburg, 79104 Freiburg, Germany Edited by William F. DeGrado, School of Pharmacy, University of California, San Francisco, CA, and approved November 29, 2016 (received for review July 28, 2016) The Bcl-2 (B-cell lymphoma 2) protein Bax (Bcl-2 associated X, across bilayers via changes in the oligomeric state or protein con- apoptosis regulator) can commit cells to apoptosis via outer mito- formation (22–24). The Bax TMD targets fusion proteins to the chondrial membrane permeabilization. Bax activity is controlled in OMM; its deletion results in cytosolic Bax localization and impaired healthy cells by prosurvival Bcl-2 proteins. C-terminal Bax trans- Bax activity (25). Analysis of the active Bax membrane topology membrane domain interactions were implicated recently in Bax pore suggests that the TMD could play a central role in Bax oligomeri- formation.
    [Show full text]
  • Cordycepin Induces Apoptosis in Human Liver Cancer Hepg2 Cells Through Extrinsic and Intrinsic Signaling Pathways
    ONCOLOGY LETTERS 12: 995-1000, 2016 Cordycepin induces apoptosis in human liver cancer HepG2 cells through extrinsic and intrinsic signaling pathways LE-WEN SHAO1, LI-HUA HUANG1, SHENG YAN2, JIAN-DI JIN3 and SHAO-YAN REN3 1Nursing Department; Departments of 2Hepato-Biliary-Pancreatic Surgery and 3Infectious Disease, The First Affiliated Hospital, College of Medicine, Zhejiang University, Hangzhou, Zhejiang 310003, P.R. China Received March 6, 2015; Accepted April 12, 2016 DOI: 10.3892/ol.2016.4706 Abstract. Cordycepin, also termed 3'-deoxyadenosine, is a to have natural medicinal properties, such as anti-angiogenic, nucleoside analogue from Cordyceps sinensis and has been anti‑tumor, anti‑viral, anti‑inflammatory and hypoglycemic reported to demonstrate numerous biological and pharma- effects (1-8). cological properties. Our previous study illustrated that the Cordycepin, also termed 3'-deoxyadenosine, is a nucleo- anti-tumor effect of cordycepin may be associated with apop- side analogue from C. militaris (9) and has been reported tosis. In the present study, the apoptotic effect of cordycepin to demonstrate numerous notable biological and pharma- on HepG2 cells was investigated using 4',6-diamidino-2-phe- cological properties, including immunological stimulation, nylindole, tetraethylbenzimidazolylcarbocyanine iodide and anti-cancer and anti-viral effects (10-15), a stimulating effect propidium iodide staining analysis and flow cytometry. The on interlukin-10 production as an immune modulator (16) and results showed that cordycepin exhibited the ability to inhibit preventing hyperlipidermia (17). HepG2 cells in a time- and dose-dependent manner when cells Apoptosis, also termed programmed cell death, is a key produced typical apoptotic morphological changes, including regulator of tissue homeostasis and is characterized by chromatin condensation, the accumulation of sub-G1 cells and typical morphological and biochemical hallmarks, including change mitochondrial permeability.
    [Show full text]
  • Caspase-6 Induces 7A6 Antigen Localization to Mitochondria During FAS-Induced Apoptosis of Jurkat Cells HIROAKI SUITA, TAKAHISA SHINOMIYA and YUKITOSHI NAGAHARA
    ANTICANCER RESEARCH 37 : 1697-1704 (2017) doi:10.21873/anticanres.11501 Caspase-6 Induces 7A6 Antigen Localization to Mitochondria During FAS-induced Apoptosis of Jurkat Cells HIROAKI SUITA, TAKAHISA SHINOMIYA and YUKITOSHI NAGAHARA Division of Life Science and Engineering, School of Science and Engineering, Tokyo Denki University, Hatoyama, Japan Abstract. Background: Mitochondria are central to caspases (caspase-8 and -9) that activate effector caspases (1). apoptosis. However, apoptosis progression involving Caspases are constructed of a pro-domain, a large subunit, and mitochondria is not fully understood. A factor involved in a small subunit. Upon caspase activation, cleavage of the mitochondria-mediated apoptosis is 7A6 antigen. 7A6 caspase occurs. Caspase-6 and -7, which are effector caspases, localizes to mitochondria from the cytosol during apoptosis, each have a large subunit that is 20 kDa (p20) and a small which seems to involve ‘effector’ caspases. In this study, we subunit that is 10 kDa (p10), but caspase-3, which is also an investigated the precise role of effector caspases in 7A6 effector caspase, has a large subunit of 17 kDa and a small localization to mitochondria during apoptosis. Materials and subunit of 12 kDa (p12) (2-4). Moreover, the large and small Methods: Human T-cell lymphoma Jurkat cells were treated subunits of caspase-6 and -7 are connected by a linker, but with an antibody against FAS. 7A6 localization was analyzed caspase-3 does not have a linker (Figure 1). The difference by confocal laser scanning microscopy and flow cytometry. between effector and initiator caspases is that the pro-domain Caspases activation was determined by western blot of an initiator caspase is long and that the pro-domain of an analysis.
    [Show full text]
  • Caspase Substrates and Inhibitors
    Downloaded from http://cshperspectives.cshlp.org/ on September 27, 2021 - Published by Cold Spring Harbor Laboratory Press Caspase Substrates and Inhibitors Marcin Pore˛ba1, Aleksandra Stro´z˙yk1, Guy S. Salvesen2, and Marcin Dra˛g1 1Department of Bioorganic Chemistry, Faculty of Chemistry, Wroclaw University of Technology, 50-370 Wrocław, Poland 2Program in Cell Death Research, Sanford-Burnham Medical Research Institute, La Jolla, California 92024 Correspondence: [email protected] Caspases are proteases at the heart of networks that govern apoptosis and inflammation. The past decade has seen huge leaps in understanding the biology and chemistry of the caspases, largely through the development of synthetic substrates and inhibitors. Such agents are used to define the role of caspases in transmitting life and death signals, in imaging caspases in situ and in vivo, and in deconvoluting the networks that govern cell behavior. Additionally, focused proteomics methods have begun to reveal the natural substrates of caspases in the thousands. Together, these chemical and proteomics technologies are setting the scene for designing and implementing control of caspase activity as appropriate targets for disease therapy. WHAT IT TAKES TO BE A CASPASE group, known as cysteine protease clan CD that he need to recycle scarce amino acids in early also contains the plant metacaspases (Vercam- Tbiotic environments required that proteases men et al. 2004), and mammalian and plant were among the first proteins to appear in the proteases of the legumain family (involved in most primitive replicating organisms. Since that specific processing events), the eukaryotic pro- distant origin, proteases have diversified into tease separase (required for sisterchromatid sep- every biological niche, occupied every cellular aration during mitosis), and the bacterial prote- and extracellular compartment, and are encod- ases clostripain and gingipain (Chen et al.
    [Show full text]
  • Proteolytic Cleavage—Mechanisms, Function
    Review Cite This: Chem. Rev. 2018, 118, 1137−1168 pubs.acs.org/CR Proteolytic CleavageMechanisms, Function, and “Omic” Approaches for a Near-Ubiquitous Posttranslational Modification Theo Klein,†,⊥ Ulrich Eckhard,†,§ Antoine Dufour,†,¶ Nestor Solis,† and Christopher M. Overall*,†,‡ † ‡ Life Sciences Institute, Department of Oral Biological and Medical Sciences, and Department of Biochemistry and Molecular Biology, University of British Columbia, Vancouver, British Columbia V6T 1Z4, Canada ABSTRACT: Proteases enzymatically hydrolyze peptide bonds in substrate proteins, resulting in a widespread, irreversible posttranslational modification of the protein’s structure and biological function. Often regarded as a mere degradative mechanism in destruction of proteins or turnover in maintaining physiological homeostasis, recent research in the field of degradomics has led to the recognition of two main yet unexpected concepts. First, that targeted, limited proteolytic cleavage events by a wide repertoire of proteases are pivotal regulators of most, if not all, physiological and pathological processes. Second, an unexpected in vivo abundance of stable cleaved proteins revealed pervasive, functionally relevant protein processing in normal and diseased tissuefrom 40 to 70% of proteins also occur in vivo as distinct stable proteoforms with undocumented N- or C- termini, meaning these proteoforms are stable functional cleavage products, most with unknown functional implications. In this Review, we discuss the structural biology aspects and mechanisms
    [Show full text]
  • Intertwined Functions of Separase and Caspase in Cell Division
    bioRxiv preprint doi: https://doi.org/10.1101/653584; this version posted May 30, 2019. The copyright holder for this preprint (which was not certified by peer review) is the author/funder. All rights reserved. No reuse allowed without permission. Intertwined Functions of Separase and Caspase in Cell Division and Programmed Cell Death Pan-Young Jeong1, Ashish Kumar1, Pradeep Joshi1, and Joel H. Rothman1* Affiliations: 1Department of Molecular, Cellular, and Developmental Biology, and Neuroscience Research Institute, University of California, Santa Barbara, Santa Barbara, CA 93106, USA. *Correspondence to: [email protected] Key words : C. elegans, chromosome separation, separase, caspase, programmed cell death 2 bioRxiv preprint doi: https://doi.org/10.1101/653584; this version posted May 30, 2019. The copyright holder for this preprint (which was not certified by peer review) is the author/funder. All rights reserved. No reuse allowed without permission. Abstract: Timely sister chromatid separation, promoted by separase, is essential for faithful chromosome segregation. Separase is a member of the CD clan of cysteine proteases, which also includes the pro-apoptotic enzymes known as caspases. We report that the C. elegans separase SEP-1, primarily known for its role in cell division, is required for apoptosis when the predominant pro-apoptotic caspase CED-3 is compromised. Loss of SEP-1 results in extra surviving cells in a weak ced-3(-) mutant, and suppresses the embryonic lethality of a mutant defective for the apoptotic suppressor ced-9/Bcl-2. We also report apparent non-apoptotic roles for CED-3 in promoting germ cell proliferation and germline meiotic chromosome disjunction and the normal rate of embryonic development.
    [Show full text]
  • Caspase-Independent Photoreceptor Apoptosis in Vivo and Differential Expression of Apoptotic Protease Activating Factor-1 and Caspase-3 During Retinal Development
    Cell Death and Differentiation (2002) 9, 1220 ± 1231 ã 2002 Nature Publishing Group All rights reserved 1350-9047/02 $25.00 www.nature.com/cdd Caspase-independent photoreceptor apoptosis in vivo and differential expression of apoptotic protease activating factor-1 and caspase-3 during retinal development M Donovan1 and TG Cotter*,1 Glu-Val-Asp-¯uromethylketone; DMSO, dimethyl sulphoxide; FITC, ¯uorescein isothiocyanate; GAPDH, glyceraldehydes-3- 1 Tumour Biology Laboratory, Department of Biochemistry, Lee Maltings, phosphate dehydrogenase; INL, inner nuclear layer; ip, intraper- University College Cork, Cork, Ireland itoneal; ONL, outer nuclear layer; PI3K, phosphatidylinositol 3- * Corresponding author: Department of Biochemistry, Lee Maltings, University kinase; PBS, phosphate buffered saline; RCS, Royal College of College Cork, Cork, Ireland. Tel:353-21-4904068; Fax: 353-21-4904259; Surgeons; rd, retinal degeneration; RP, Retinitis Pigmentosa; E-mail: [email protected][email protected] TUNEL, Terminal dUTP nick end labelling; TdT, terminal deox- Received 7.5.02; revised 3.7.02; accepted 5.7.02 ynucleotidyl transferase; UV, ultraviolet; zVAD-fmk, Z-Val-Ala- Edited by G Ciliberto Asp.¯uoromethylketone Abstract Introduction Apoptosis is the mode of photoreceptor cell death in many Light-induced photoreceptor apoptosis represents an animal retinal dystrophies. Exposure of Balb/c mice to excessive model for the study of retinal degenerations such as Retinitis levels of light induces photoreceptor apoptosis and repre- Pigmentosa (RP), a genetically diverse group of disorders sents an animal model for the study of retinal degenerations. involving the progressive death of photoreceptor cells.1 Caspases have emerged as central regulators of apoptosis, Apoptosis is the mode of photoreceptor cell death in both executing this tightly controlled death pathway in many cells.
    [Show full text]
  • Roles of Bid and XIAP
    Cell Death and Differentiation (2012) 19, 42–50 & 2012 Macmillan Publishers Limited All rights reserved 1350-9047/12 www.nature.com/cdd Review Fas death receptor signalling: roles of Bid and XIAP T Kaufmann*,1, A Strasser2,3 and PJ Jost4 Fas (also called CD95 or APO-1), a member of a subgroup of the tumour necrosis factor receptor superfamily that contain an intracellular death domain, can initiate apoptosis signalling and has a critical role in the regulation of the immune system. Fas-induced apoptosis requires recruitment and activation of the initiator caspase, caspase-8 (in humans also caspase-10), within the death-inducing signalling complex. In so-called type 1 cells, proteolytic activation of effector caspases (-3 and -7) by caspase-8 suffices for efficient apoptosis induction. In so-called type 2 cells, however, killing requires amplification of the caspase cascade. This can be achieved through caspase-8-mediated proteolytic activation of the pro-apoptotic Bcl-2 homology domain (BH)3-only protein BH3-interacting domain death agonist (Bid), which then causes mitochondrial outer membrane permeabilisation. This in turn leads to mitochondrial release of apoptogenic proteins, such as cytochrome c and, pertinent for Fas death receptor (DR)-induced apoptosis, Smac/DIABLO (second mitochondria-derived activator of caspase/direct IAP binding protein with low Pi), an antagonist of X-linked inhibitor of apoptosis (XIAP), which imposes a brake on effector caspases. In this review, written in honour of Juerg Tschopp who contributed so much to research on cell death and immunology, we discuss the functions of Bid and XIAP in the control of Fas DR-induced apoptosis signalling, and we speculate on how this knowledge could be exploited to develop novel regimes for treatment of cancer.
    [Show full text]