DOCSLIB.ORG

General Event in the Execution of Apoptosis a Proinflammatory Signal

Total Page：16

File Type：pdf, Size：1020Kb

Download full-text PDF Read full-text
General Event in the Execution of Apoptosis a Proinflammatory Signal Cell Death Triggered by Yersinia enterocolitica Identifies Processing of the Proinflammatory Signal Adapter MyD88 as a General Event in the Execution of Apoptosis This information is current as of September 30, 2021. Lena Novikova, Nicole Czymmeck, Anne Deuretzbacher, Friedrich Buck, Kathleen Richter, Alexander N. R. Weber, Martin Aepfelbacher and Klaus Ruckdeschel J Immunol published online 20 December 2013 http://www.jimmunol.org/content/early/2013/12/20/jimmun Downloaded from ol.1203464 References This article cites 40 articles, 15 of which you can access for free at: http://www.jimmunol.org/ http://www.jimmunol.org/content/early/2013/12/20/jimmunol.1203464.ful l#ref-list-1 Why The JI? Submit online. • Rapid Reviews! 30 days* from submission to initial decision • No Triage! Every submission reviewed by practicing scientists by guest on September 30, 2021 • Fast Publication! 4 weeks from acceptance to publication *average 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 © 2013 by The American Association of Immunologists, Inc. All rights reserved. Print ISSN: 0022-1767 Online ISSN: 1550-6606. Published December 20, 2013, doi:10.4049/jimmunol.1203464 The Journal of Immunology Cell Death Triggered by Yersinia enterocolitica Identifies Processing of the Proinflammatory Signal Adapter MyD88 as a General Event in the Execution of Apoptosis Lena Novikova,* Nicole Czymmeck,* Anne Deuretzbacher,*,† Friedrich Buck,‡ Kathleen Richter,x,{ Alexander N. R. Weber,‖ Martin Aepfelbacher,* and Klaus Ruckdeschel*,x Many pathogenic microorganisms have evolved tactics to modulate host cell death or survival pathways for establishing infection. The enteropathogenic bacterium Yersinia enterocolitica deactivates TLR-induced signaling pathways, which triggers apoptosis in macrophages. In this article, we show that Yersinia-induced apoptosis of human macrophages involves caspase-dependent cleavage of the TLR adapter protein MyD88. MyD88 was also cleaved when apoptosis was mediated by overexpression of the Toll–IL-1R Downloaded from domain–containing adapter inducing IFN-b in epithelial cells. The caspase-processing site was mapped to aspartate-135 in the central region of MyD88. MyD88 is consequently split by caspases in two fragments, one harboring the death domain and the other the Toll–IL-1R domain. Caspase-3 was identified as the protease that conferred the cleavage of MyD88 in in vitro caspase assays. In line with a broad role of caspase-3 in the execution of apoptosis, the processing of MyD88 was not restricted to Yersinia infection and to proapoptotic Toll–IL-1R domain–containing adapter inducing IFN-b signaling, but was also triggered by staur- osporine treatment. The cleavage of MyD88 therefore seems to be a common event in the advanced stages of apoptosis, when http://www.jimmunol.org/ caspase-3 is active. We propose that the processing of MyD88 disrupts its scaffolding function and uncouples the activation of TLR and IL-1Rs from the initiation of proinflammatory signaling events. The disruption of MyD88 may consequently render dying cells less sensitive to proinflammatory stimuli in the execution phase of apoptosis. The cleavage of MyD88 could therefore be a means of conferring immunogenic tolerance to apoptotic cells to ensure silent, noninflammatory cell demise. The Journal of Immunology, 2014, 192: 000–000. oll-like receptors are transmembrane receptors that play typic TIR–TIR domain interaction (3). MyD88 is a central TIR an essential role in the recognition of foreign, invading adapter that is implicated in TLR and IL-1R signaling (4). It is by guest on September 30, 2021 T microbial pathogens. They are activated by conserved, activated by the IL-1R family and by all TLRs with the exception pathogen-associated molecular patterns and trigger the induction of of TLR3. In contrast, TLR3, stimulated by microbial dsRNA, protective proinflammatory signaling pathways (1, 2). The human signals via the Toll–IL-1R domain–containing adapter inducing TLR family comprises . 10 TLRs, each detecting a specific set of IFN-b (TRIF). TRIF is recruited also by TLR4, which is the sole ligands. LPS from Gram-negative bacteria, for instance, activates TLR that can engage both the MyD88 and TRIF pathways. The TLR4, whereas bacterial lipoproteins signal through TLR2. The TIR adapter protein–dependent signals converge on NF-kB and TLR-triggered effects depend on the activation of signaling cas- MAPK activation, which mounts the inflammatory response. Mean- cades that are initiated by the recruitment of intracytoplasmic while, it is well established that the TLRs, in addition to promoting adapter proteins to the Toll–IL-1R (TIR) domain through homo- inflammation, are also implicated in controlling cellular life and death. Hence, the engagement of TLRs may enable cell survival, but may also elicit cell death through either apoptosis or necrosis *Institute for Medical Microbiology, Virology and Hygiene, University Medical (5, 6). The signaling networks that control the differential out- † Center Eppendorf, 20246 Hamburg, Germany; Division of Hematology, Department come are not yet completely understood. However, it seems that of Internal Medicine II, University Hospital of Wu¨rzburg, 97080 Wu¨rzburg, Ger- many; ‡Institute of Clinical Chemistry, University Medical Center Eppendorf, 20246 the type of cell, as well as the composition of the signals induced x Hamburg, Germany; Max von Pettenkofer-Institute for Hygiene and Medical Mi- by the microbial challenge, determines the fate of the stimulated crobiology, 80336 Munich, Germany; {Stiefel/GlaxoSmithKline Early Development, ‖ Durham, NC 27709; and Department of Immunology, University of Tu¨bingen, cell. 72076 Tu¨bingen, Germany Several pathogenic bacteria have evolved mechanisms to mod- Received for publication December 19, 2012. Accepted for publication November ify TLR signaling pathways for establishing infection (7, 8). One 20, 2013. example is given by the Gram-negative bacterial pathogen Yersi- This work was supported by Deutsche Forschungsgemeinschaft Grants Ru788/3-1, 3-2. nia, which deregulates TLR-induced immune responses by en- Address correspondence and reprint requests to Dr. Klaus Ruckdeschel, Institute for gaging Yersinia outer protein (Yop) P in Y. enterocolitica, or the Medical Microbiology, Virology and Hygiene, University Medical Center Eppendorf, homologous YopJ protein in Y. pestis and Y. pseudotuberculosis. Martinistrasse 52, 20246 Hamburg, Germany. E-mail address: [email protected] YopP/YopJ is injected with several other Yops by Yersinia type III Abbreviations used in this article: DD, death domain; HEK, human embryonic kid- ney; IKK, inhibitory kB kinase; IRAK, IL-1R–associated kinase; MOI, multiplicity protein secretion into host cells to modify critical signaling path- of infection; PVDF, polyvinylidene difluoride; TAK1, TGF-b–activated kinase-1; ways of innate immunity (9, 10). YopP/YopJ acts as an acetyltrans- TIR, Toll–IL-1R; TRIF, Toll–IL-1R domain–containing adapter inducing IFN-b; ferase on the NF-kB–activating IkB kinase (IKK) b and on mem- Yop, Yersinia outer protein. bers of the MAPK kinase superfamily (11, 12). The acetylation of Copyright Ó 2013 by The American Association of Immunologists, Inc. 0022-1767/13/$16.00 these kinases deactivates the NF-kB and MAPK pathways and www.jimmunol.org/cgi/doi/10.4049/jimmunol.1203464 2 TARGETING AND REPROGRAMMING OF MyD88 IN APOPTOSIS downregulates the proinflammatory host cell response. Because micin (100 mg/ml) after 90 min of infection to prevent bacterial over- these pathways also promote cell survival, the action of YopP/YopJ growth. Staurosporine (Sigma-Aldrich, Munich, Germany) was applied sensitizes macrophages to undergo apoptosis following Yersinia to the cells at a concentration of 5 mM. Where indicated, the cells were pretreated with 40 mM of the caspase inhibitor zVAD-fmk (Bachem, infection (13, 14). The proapoptotic response thereby is accelerated Bubendorf, Switzerland) 30 min prior to infection. by the activation of TLR4 and TRIF, whereas the MyD88 pathway Eukaryotic expression plasmids, cell transfection, and seems to be dispensable for Yersinia-induced cell death (15, 16). measurement of NF-kB activation TRIF is connected to the cell death machinery by the adapter’s receptor-interacting protein 1 and Fas-associated death domain Transfections of HEK293 cells were performed by the calcium phosphate (DD) to initiate apoptosis through caspase-8 (16–18). The signal- method using ProFection according to the manufacturer’s instructions (Promega, Mannheim, Germany). For the detection of transiently over- ing of apoptosis following TLR4 or TLR3 activation seems to be expressed Myc-tagged MyD88 (24), the cells were transfected with 3 mg a prominent function of TRIF, which is substantiated by the finding expression plasmid in 6-cm cell culture dishes. Where indicated, plas- that the overexpression of TRIF significantly compels transfected mids for TRIF (25), the caspase-8 inhibitor CrmA (26), or IL-1R–as- cells to undergo apoptosis (16–18). sociated kinase (IRAK) 4 (27) were included.
Load more

Recommended publications
  • Role of CCCH-Type Zinc Finger Proteins in Human Adenovirus Infections
    viruses Review Role of CCCH-Type Zinc Finger Proteins in Human Adenovirus Infections Zamaneh Hajikhezri 1, Mahmoud Darweesh 1,2, Göran Akusjärvi 1 and Tanel Punga 1,* 1 Department of Medical Biochemistry and Microbiology, Uppsala University, 75123 Uppsala, Sweden; [email protected] (Z.H.); [email protected] (M.D.); [email protected] (G.A.) 2 Department of Microbiology and Immunology, Al-Azhr University, Assiut 11651, Egypt * Correspondence: [email protected]; Tel.: +46-733-203-095 Received: 28 October 2020; Accepted: 16 November 2020; Published: 18 November 2020 Abstract: The zinc finger proteins make up a significant part of the proteome and perform a huge variety of functions in the cell. The CCCH-type zinc finger proteins have gained attention due to their unusual ability to interact with RNA and thereby control different steps of RNA metabolism. Since virus infections interfere with RNA metabolism, dynamic changes in the CCCH-type zinc finger proteins and virus replication are expected to happen. In the present review, we will discuss how three CCCH-type zinc finger proteins, ZC3H11A, MKRN1, and U2AF1, interfere with human adenovirus replication. We will summarize the functions of these three cellular proteins and focus on their potential pro- or anti-viral activities during a lytic human adenovirus infection. Keywords: human adenovirus; zinc finger protein; CCCH-type; ZC3H11A; MKRN1; U2AF1 1. Zinc Finger Proteins Zinc finger proteins are a big family of proteins with characteristic zinc finger (ZnF) domains present in the protein sequence. The ZnF domains consists of various ZnF motifs, which are short 30–100 amino acid sequences, coordinating zinc ions (Zn2+).
    [Show full text]
  • TRAF5, a Novel Tumor Necrosis Factor Receptor-Associated Factor Family
    Proc. Natl. Acad. Sci. USA Vol. 93, pp. 9437-9442, September 1996 Biochemistry TRAF5, a novel tumor necrosis factor receptor-associated factor family protein, mediates CD40 signaling (signal transduction/protein-protein interaction/yeast two-hybrid system) TAKAoMI ISHIDA*, TADASHI ToJo*, TSUTOMU AOKI*, NORIHIKO KOBAYASHI*, TSUKASA OHISHI*, TOSHIKI WATANABEt, TADASHI YAMAMOTO*, AND JUN-ICHIRO INOUE*t Departments of *Oncology and tPathology, The Institute of Medical Science, The University of Tokyo, 4-6-1 Shirokanedai, Minato-ku, Tokyo 108, Japan Communicated by David Baltimore, Massachusetts Institute of Technology, Cambridge, MA, May 22, 1996 (received for review March 8, 1996) ABSTRACT Signals emanating from CD40 play crucial called a death domain, suggesting that these receptors could roles in B-cell function. To identify molecules that transduce have either common or similar signaling mechanisms (13). CD40 signalings, we have used the yeast two-hybrid system to Biochemical purification of receptor-associated proteins or the clone cDNAs encoding proteins that bind the cytoplasmic tail recently developed cDNA cloning system that uses yeast of CD40. A cDNA encoding a putative signal transducer genetic selection led to the discovery of two groups of signal protein, designated TRAF5, has been molecularly cloned. transducer molecules. Members of the first group are proteins TRAF5 has a tumor necrosis factor receptor-associated factor with a TRAF domain for TNFR2 and CD40 such as TRAF1, (TRAF) domain in its carboxyl terminus and is most homol- TRAF2 (17), and TRAF3, also known as CD40bp, LAP-1, or ogous to TRAF3, also known as CRAF1, CD40bp, or LAP-1, CRAF1 or CD40 receptor-associated factor (18-20).
    [Show full text]
  • FLIP and the Death Effector Domain Family
    Oncogene (2008) 27, 6216–6227 & 2008 Macmillan Publishers Limited All rights reserved 0950-9232/08 $32.00 www.nature.com/onc REVIEW FLIP and the death effector domain family JW Yu and Y Shi Department of Molecular Biology, Lewis Thomas Laboratory, Princeton University, Princeton, NJ, USA Death effector domains (DEDs) are protein interaction and stress that impinge on the mitochondria or modules found in a number of proteins known to regulate ‘extrinsically’ from extracellular ligands that activate apoptosis from death receptors. The core DED family death receptors at the cell surface. In either scenario, members that orchestrate programmed cell death from each pathway critically relies on the action of a family of death receptors include the adaptor protein FADD, the cysteine proteases known as caspases to initiate and initiator caspases procaspases-8 and -10 and the regulatory execute the cell death process through a two-step protein c-FLIP. Through homotypic DED interactions, cascade (Riedl and Shi, 2004). In the apoptotic these proteins assemble into the death-inducingsignaling proteolytic cascade, the initiator caspases (caspases-2, complex (DISC) to regulate initiator caspase activation -8, -9 and -10) cleave and consequently activate the and launch the apoptotic proteolytic cascade. A consider- effector caspases (caspases-3, -6 and -7), and the effector able body of evidence, however, is revealingthat the same caspases in turn cleave a large array of substrates to core group of DED-containing proteins also paradoxically dismantle and package the cell into apoptotic bodies. promotes survival and proliferation in lymphocytes and Activation of initiator caspases for both the intrinsic possibly other cell types.
    [Show full text]
  • Purification, Crystallization and Preliminary X-Ray Crystallographic Studies of RAIDD Death-Domain (DD)
    Int. J. Mol. Sci. 2009, 10, 2501-2509; doi:10.3390/ijms10062501 OPEN ACCESS International Journal of Molecular Sciences ISSN 1422-0067 www.mdpi.com/journal/ijms Article Purification, Crystallization and Preliminary X-ray Crystallographic Studies of RAIDD Death-Domain (DD) Tae-ho Jang and Hyun Ho Park * Department of Biochemistry, School of Biotechnology at Yeungnam University, Gyeong-san, Korea * Author to whom correspondence should be addressed; E-Mail: [email protected]; Tel. +1-82-53-810-3045; Fax: +1-82-53-810-4769 Received: 7 April 2009; in revised form: 26 May 2009 / Accepted: 1 June 2009 / Published: 3 June 2009 Abstract: Caspase-2 activation by formation of PIDDosome is critical for genotoxic stress induced apoptosis. PIDDosome is composed of three proteins, RAIDD, PIDD, and Caspase-2. RAIDD is an adaptor protein containing an N-terminal Caspase-Recruiting- Domain (CARD) and a C-terminal Death-Domain (DD). Its interactions with Caspase-2 and PIDD through CARD and DD respectively and formation of PIDDosome are important for the activation of Caspase-2. RAIDD DD cloned into pET26b vector was expressed in E. coli cells and purified by nickel affinity chromatography and gel filtration. Although it has been known that the most DDs are not soluble in physiological condition, RAIDD DD was soluble and interacts tightly with PIDD DD in physiological condition. The purified RAIDD DD alone has been crystallized. Crystals are trigonal and belong to space group P3121 (or its enantiomorph P3221) with unit-cell parameters a = 56.3, b = 56.3, c = 64.9 Å and γ = 120°.
    [Show full text]
  • PARP13 Regulates Cellular Mrna Post-Transcriptionally and Functions As a Pro-Apoptotic Factor by Destabilizing TRAILR4 Transcript
    ARTICLE Received 4 Sep 2014 | Accepted 23 Sep 2014 | Published 10 Nov 2014 DOI: 10.1038/ncomms6362 PARP13 regulates cellular mRNA post-transcriptionally and functions as a pro-apoptotic factor by destabilizing TRAILR4 transcript Tanya Todorova1,2, Florian J. Bock2 & Paul Chang1,2 Poly(ADP-ribose) polymerase-13 (PARP13/ZAP/ZC3HAV1) is an antiviral factor, active against specific RNA viruses such as murine leukaemia virus, Sindbis virus and human immunodeficiency virus. During infection, PARP13 binds viral RNA via its four CCCH-type zinc-finger domains and targets it for degradation by recruiting cellular messenger RNA (mRNA) decay factors such as the exosome complex and XRN1. Here we show that PARP13 binds to and regulates cellular mRNAs in the absence of viral infection. Knockdown of PARP13 results in the misregulation of hundreds of transcripts. Among the most upregulated transcripts is TRAILR4 that encodes a decoy receptor for TRAIL—a pro-apoptotic cytokine that is a promising target for the therapeutic inhibition of cancers. PARP13 destabilizes TRAILR4 mRNA post-transcriptionally in an exosome-dependent manner by binding to a region in its 30 untranslated region. As a consequence, PARP13 represses TRAILR4 expression and increases cell sensitivity to TRAIL-mediated apoptosis, acting as a key regulator of the cellular response to TRAIL. 1 Department of Biology, Massachusetts Institute of Technology, 500 Main Street, Cambridge, Massachusetts 02139, USA. 2 Koch Institute for Integrative Cancer Research, Massachusetts Institute of Technology, 500 Main Street, Cambridge, Massachusetts 02139, USA. Correspondence and requests for materials should be addressed to P.C. (email: [email protected]). NATURE COMMUNICATIONS | 5:5362 | DOI: 10.1038/ncomms6362 | www.nature.com/naturecommunications 1 & 2014 Macmillan Publishers Limited.
    [Show full text]
  • Nuclear Factor- B-Inducible Death Effector Domain-Containing Protein
    THE JOURNAL OF BIOLOGICAL CHEMISTRY Vol. 276, No. 28, Issue of July 13, pp. 26398–26404, 2001 © 2001 by The American Society for Biochemistry and Molecular Biology, Inc. Printed in U.S.A. Nuclear Factor-␬B-inducible Death Effector Domain-containing Protein Suppresses Tumor Necrosis Factor-mediated Apoptosis by Inhibiting Caspase-8 Activity* Received for publication, March 19, 2001, and in revised form, May 8, 2001 Published, JBC Papers in Press, May 9, 2001, DOI 10.1074/jbc.M102464200 Zongbing You‡, Hongjiao Ouyang‡§, Dennis Lopatin¶, Peter J. Polverʈ, and Cun-Yu Wang‡** From the ‡Laboratory of Molecular Signaling and Apoptosis, the Department of Biologic and Materials Sciences, School of Dentistry, Program in Cellular and Molecular Biology, and Comprehensive Cancer Center, the §Department of Cardiology, Restorative Sciences and Endodontics, and the ¶Department of Biologic and Materials Sciences, University of Michigan, Ann Arbor, Michigan 48109 and the ʈSchool of Dentistry, University of Minnesota, Minneapolis, Minnesota 55455 Activation of the transcription factor nuclear fac- ing pathway or trigger the caspase cascade by interacting with tor-␬B (NF-␬B) has been found to play an essential role FADD (Fas-associated death domain protein) in the absence of in the inhibition of tumor necrosis factor (TNF)-medi- NF-␬B activation (1–3). FADD recruits and activates ated apoptosis. NF-␬B regulates several antiapoptotic caspase-8. Both caspase-8 and FADD contain death effector molecules including inhibitors of apoptosis, Bcl-2 family domains (DED) that were found to play a critical role for the proteins (A1 and Bcl-XL), and IEX-IL. Here we report protein-protein interaction and caspase activation (4, 5).
    [Show full text]
  • The Death Domain Superfamily in Intracellular Signaling of Apoptosis and Inflammation
    ANRV306-IY25-19 ARI 11 February 2007 12:51 The Death Domain Superfamily in Intracellular Signaling of Apoptosis and Inflammation Hyun Ho Park,1 Yu-Chih Lo,1 Su-Chang Lin,1 Liwei Wang,1 Jin Kuk Yang,1,2 and Hao Wu1 1Department of Biochemistry, Weill Medical College and Graduate School of Medical Sciences of Cornell University, New York, New York 10021; email: [email protected] 2Department of Chemistry, Soongsil University, Seoul 156-743, Korea Annu. Rev. Immunol. 2007. 25:561–86 Key Words First published online as a Review in Advance on death domain (DD), death effector domain (DED), tandem DED, January 2, 2007 caspase recruitment domain (CARD), pyrin domain (PYD), crystal The Annual Review of Immunology is online at structure, NMR structure immunol.annualreviews.org This article’s doi: Abstract 10.1146/annurev.immunol.25.022106.141656 The death domain (DD) superfamily comprising the death domain Copyright c 2007 by Annual Reviews. (DD) subfamily, the death effector domain (DED) subfamily, the Annu. Rev. Immunol. 2007.25:561-586. Downloaded from arjournals.annualreviews.org by CORNELL UNIVERSITY MEDICAL COLLEGE on 03/29/07. For personal use only. All rights reserved caspase recruitment domain (CARD) subfamily, and the pyrin do- 0732-0582/07/0423-0561$20.00 main (PYD) subfamily is one of the largest domain superfamilies. By mediating homotypic interactions within each domain subfam- ily, these proteins play important roles in the assembly and activation of apoptotic and inflammatory complexes. In this chapter, we review the molecular complexes assembled by these proteins, the structural and biochemical features of these domains, and the molecular in- teractions mediated by them.
    [Show full text]
  • Identification of a Zinc Finger Protein Whose Subcellular Distribution Is Regulated by Serum and Nerve Growth Factor
    Proc. Natl. Acad. Sci. USA Vol. 96, pp. 10705–10710, September 1999 Cell Biology Identification of a zinc finger protein whose subcellular distribution is regulated by serum and nerve growth factor ALEXANDRA CHITTKA*† AND MOSES V. CHAO*‡ *Cell Biology Program, Weill Graduate School of Cornell University Medical College, New York, NY 10021, Skirball Institute of Biomolecular Medicine, New York University School of Medicine, New York, NY 10016 Edited by Michael V. L. Bennett, Albert Einstein College of Medicine, Woods Hole, MA, and approved July 13, 1999 (received for review March 24, 1999) ABSTRACT A subclass of zinc finger proteins containing a 18). Here, we describe the structural features of SC-1, its asso- unique protein motif called the positive regulatory (PR) domain ciation with p75 neurotrophin receptor, and its subcellular local- has been described. The members include the PRDI-BF1͞ ization. We find the distribution of SC-1 is strongly regulated by Blimp-1 protein, the Caenorhabditis elegans egl-43 and EVI1 gene NGF binding to the p75 receptor and by growth conditions. products, and the retinoblastoma interacting protein RIZ. Here Interestingly, the localization of SC-1 is not under control of NGF we describe a member of this family, SC-1, that exhibits several binding to the TrkA receptor or by other neurotrophins, such as distinctive features. First, SC-1 interacts with the p75 neurotro- brain-derived neurotrophic factor and neurotrophin 3. These phin receptor and is redistributed from the cytoplasm to the observations define SC-1 as a protein that is differentially con- nucleus after nerve growth factor (NGF) treatment of trans- trolled by neurotrophin and serum conditions.
    [Show full text]
  • Characterization of Bovine FAS-Associated Death Domain Gene1
    SHORT COMMUNICATION doi:10.1111/j.1365-2052.2004.01207.x Characterization of bovine FAS-associated death domain gene1 M. E. Szperka*, E. E. Connor*, M. J. Paape*, J. L. Williams† and D. D. Bannerman* *Bovine Functional Genomics Laboratory, USDA Agricultural Research Service, Beltsville, MD 20705, USA. †Roslin Institute, Roslin, Midlothian EH25 9PS, Edinburgh, UK Summary The FAS-associated death domain (FADD) protein is an adapter/signaling molecule that has been shown to function in human cells to promote apoptosis and to inhibit NF-jB activa- tion. Because of the critical role that apoptosis and NF-jB play in a variety of disease states, we mapped the bovine FADD gene, sequenced bovine FADD cDNA, and characterized its expression in endothelial cells (EC). Sequencing of bovine FADD revealed approximately 65 and 58% amino acid sequence identity to its human and murine homologues, respectively. Bovine FADD was mapped to chromosome 29 by radiation hybrid mapping. In addition, the functionality of bovine FADD was studied. Expression of a bovine FADD dominant-negative construct blocked bacterial lipopolysaccharide (LPS)- and TNF-a- induced apoptosis in bovine EC consistent with previous studies of human FADD. In contrast to human FADD, elevated expression of bovine FADD had no effect on LPS- or TNF- a-induced upregulation of NF-jB-dependent gene products as assayed by E-selectin expression. Thus, while the role of FADD in mediating apoptosis is conserved across species, its role in regulating NF-jB-dependent gene expression is not. Keywords apoptosis, caspase, cytokines, endotoxin, inflammation. Inflammation and apoptosis contribute to the pathogenesis enables activation of these pro-enzymes into fully functional and/or deleterious outcomes associated with many bacter- proteases resulting in the onset of apoptosis (Muzio et al.
    [Show full text]
  • Caspases: Pharmacological Manipulation of Cell Death Inna N
    Review series Caspases: pharmacological manipulation of cell death Inna N. Lavrik, Alexander Golks, and Peter H. Krammer Division of Immunogenetics, Tumor Immunology Program, German Cancer Research Center, Heidelberg, Germany. Caspases, a family of cysteine proteases, play a central role in apoptosis. During the last decade, major progress has been made to further understand caspase structure and function, providing a unique basis for drug design. This Review gives an overview of caspases and their classification, structure, and substrate specificity. We also describe the current knowledge of how interference with caspase signaling can be used to pharmacologically manipulate cell death. Introduction involved in the transduction of the apoptotic signal. This superfam- Apoptosis, or programmed cell death, is a common property of all ily consists of the death domain (DD), the death effector domain multicellular organisms (1, 2). It can be triggered by a number of (DED), and the caspase recruitment domain (CARD) (11). Each of factors, including ultraviolet or γ-irradiation, growth factor with- these motifs interacts with other proteins by homotypic interac- drawal, chemotherapeutic drugs, or signaling by death receptors tions. All members of the death domain superfamily are character- (DRs) (3, 4). The central role in the regulation and the execution of ized by similar structures that comprise 6 or 7 antiparallel amphipa- apoptotic cell death belongs to caspases (5–7). Caspases, a family thic α-helices. Structural similarity suggests a common evolutionary of cysteinyl aspartate–specific proteases, are synthesized as zymo- origin for all recruitment domains (12). However, the nature of the gens with a prodomain of variable length followed by a large sub- homotypic interactions differs within the superfamily.
    [Show full text]
  • Bid, One of the First Family Members Described with a BH3 Domain Only, Is a Cytosolic Protein and Lacks a Membrane Targeting Sequence
    Bid, one of the first family members described with a BH3 domain only, is a cytosolic protein and lacks a membrane targeting sequence. Caspase-8 cleaves Bid to yield a truncated COOH- terminal 15 kD protein (tBid), which readily moves to mitochondrial membranes. All BH3-only proteins described to date lack C-terminal membrane spanning sequences and have non-mitochondrial localizations, with mitochondrial translocation taking place downstream of specific apoptotic signaling pathways. As for Bax, changes in protein conformation may be required for membrane targeting. BH3 alignments As Bid contains a buried BH3 domain, Bid can be classified as a latent pro-apoptotic protein. The specific activation mechanism for Bid, caspase-mediated cleavage, is predicted to lead to exposure of the BH3 domain. Similarly, caspase proteolysis at the NH -terminus 2 of Bcl-2 and Bcl-x , generates pro-apoptotic versions of these proteins. The NH -terminal L 2 a-helix forms an undercarriage for the BH3 helix, thus removing this portion of the protein may untether the BH3 domains. Lipid interactions may also be involved in membrane targeting of BH3-only proteins. Membrane insertion of tBid is favored in lipid bilayers containing >20% cardiolipin, a mitochondria-specific phospholipid normally restricted to the inner mitochondrial membrane. Post-translational lipid modification of tBid takes place by myristoylation at the NH -terminal glycine generated following caspase-mediated 2 cleavage. The pro-apoptotic Bad protein also lacks a COOH-terminal signal/anchor sequence but has two consensus 14-3-3 binding sites. Phosphorylation at serines 112 and 136 within the 14-3-3 binding site results in sequestration of Bad bound to cytosolic 14- 3-3 proteins.
    [Show full text]
  • Obscurin Mediates Ankyrin Complex Formation in the Heart
    The Texas Medical Center Library DigitalCommons@TMC The University of Texas MD Anderson Cancer Center UTHealth Graduate School of The University of Texas MD Anderson Cancer Biomedical Sciences Dissertations and Theses Center UTHealth Graduate School of (Open Access) Biomedical Sciences 8-2019 OBSCURIN MEDIATES ANKYRIN COMPLEX FORMATION IN THE HEART Janani Subramaniam Follow this and additional works at: https://digitalcommons.library.tmc.edu/utgsbs_dissertations Part of the Biochemistry Commons, Integrative Biology Commons, and the Molecular Biology Commons Recommended Citation Subramaniam, Janani, "OBSCURIN MEDIATES ANKYRIN COMPLEX FORMATION IN THE HEART" (2019). The University of Texas MD Anderson Cancer Center UTHealth Graduate School of Biomedical Sciences Dissertations and Theses (Open Access). 961. https://digitalcommons.library.tmc.edu/utgsbs_dissertations/961 This Thesis (MS) is brought to you for free and open access by the The University of Texas MD Anderson Cancer Center UTHealth Graduate School of Biomedical Sciences at DigitalCommons@TMC. It has been accepted for inclusion in The University of Texas MD Anderson Cancer Center UTHealth Graduate School of Biomedical Sciences Dissertations and Theses (Open Access) by an authorized administrator of DigitalCommons@TMC. For more information, please contact [email protected]. OBSCURIN MEDIATES ANKYRIN COMPLEX FORMATION IN THE HEART by Janani Subramaniam, B.S. APPROVED: ______________________________ Shane R. Cunha, Ph.D. Advisory Professor ______________________________
    [Show full text]