DOCSLIB.ORG

Neutrophils Phagocytosis in Gelsolin-Deficient Murine Selective

Total Page：16

File Type：pdf, Size：1020Kb

Download full-text PDF Read full-text
Neutrophils Phagocytosis in Gelsolin-Deficient Murine Selective Selective Inhibition of IgG-Mediated Phagocytosis in Gelsolin-Deficient Murine Neutrophils This information is current as Lena Serrander, Petra Skarman, Birgitta Rasmussen, Walter of October 2, 2021. Witke, Daniel P. Lew, Karl-Heinz Krause, Olle Stendahl and Oliver Nüße J Immunol 2000; 165:2451-2457; ; doi: 10.4049/jimmunol.165.5.2451 http://www.jimmunol.org/content/165/5/2451 Downloaded from References This article cites 46 articles, 26 of which you can access for free at: http://www.jimmunol.org/content/165/5/2451.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 by guest on October 2, 2021 *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 © 2000 by The American Association of Immunologists All rights reserved. Print ISSN: 0022-1767 Online ISSN: 1550-6606. Selective Inhibition of IgG-Mediated Phagocytosis in Gelsolin-Deficient Murine Neutrophils1 Lena Serrander,2* Petra Skarman,† Birgitta Rasmussen,† Walter Witke,‡ Daniel P. Lew,* Karl-Heinz Krause,3* Olle Stendahl,† and Oliver Nu¨ße4* Phagocytosis and the microbicidal functions of neutrophils require dynamic changes of the actin cytoskeleton. We have investi- gated the role of gelsolin, a calcium-dependent actin severing and capping protein, in peripheral blood neutrophils from gelsolin- null (Gsn؊) mice. The phagocytosis of complement opsonized yeast was only minimally affected. In contrast, phagocytosis of IgG-opsonized yeast was reduced close to background level in Gsn؊ neutrophils. Thus, gelsolin is essential for efficient IgG- but not complement-mediated phagocytosis. Furthermore, attachment of IgG-opsonized yeast to Gsn؊ neutrophils was reduced (ϳ50%) but not to the same extent as ingestion (ϳ73%). This was not due to reduced surface expression of the Fc␥-receptor or its lateral mobility. This suggests that attachment and ingestion of IgG-opsonized yeast by murine neutrophils are actin-dependent Downloaded from and gelsolin is important for both steps in phagocytosis. We also investigated granule exocytosis and several steps in phagosome processing, namely the formation of actin around the phagosome, translocation of granules, and activation of the NADPH-oxidase. All these functions were normal in Gsn؊ neutrophils. Thus, the role of gelsolin is specific for IgG-mediated phagocytosis. Our data suggest that gelsolin is part of the molecular machinery that distinguishes complement and IgG-mediated phagocytosis. The latter requires a more dynamic reorganization of the cytoskeleton. The Journal of Immunology, 2000, 165: 2451–2457. olymorphonuclear leukocytes (neutrophils) constantly re- olites and granular enzymes into the phagosomal space, demand http://www.jimmunol.org/ arrange their actin cytoskeleton to perform chemotaxis and rapid and dynamic cytoskeletal rearrangements. Gelsolin consti- phagocytosis. Such rearrangements depend on actin sev- tutes about 1% of the total neutrophil protein and is a good can- P 2ϩ ering and capping proteins of which gelsolin is the best-character- didate to mediate Ca -dependent actin rearrangements (6). ized example (1). Gelsolin is an 82-kDa cytoplasmic protein Phagocytosis in neutrophils is well studied, but its molecular ϩ widely expressed in mammalian cells. Upon activation by Ca2 , mechanisms are still poorly understood. The two main pathways gelsolin severs actin filaments, binds, and caps barbed ends of are complement-receptor (CR)-mediated and Fc␥-receptor actin filaments. Gelsolin-actin complexes dissociate in the pres- (Fc␥R)-mediated phagocytosis (reviewed in Ref. 7). The important Ϫ 5 ence of polyphosphoinositides (2, 3). Gelsolin-null (Gsn ) mice receptors are CR1, CR3, and CR4 for CR-mediated phagocytosis by guest on October 2, 2021 Ϫ have normal embryonic development and longevity (4). Gsn and Fc␥R II and Fc␥R III for Fc␥R-mediated phagocytosis in non- mice show a prolonged bleeding time due to decreased platelet stimulated neutrophils. shape changes. Migration of neutrophils into peritoneal exudates is Binding and clustering of CRs by complement-opsonized par- delayed, and fibroblasts have excessive stress fibers and decreased Ϫ ticles mediates serine phosphorylation of the receptor and the for- motility. The GTP-binding protein rac is overexpressed in Gsn mation of punctuate structures containing F-actin, paxillin, ␣-ac- tissues and regulates gelsolin in fibroblasts (5). tinin, and vinculin assembled to the membrane (8). This leads to a Neutrophil functions, such as adhesion, chemotaxis, phagocy- sinking of the opsonized particle into the membrane without pro- tosis and finally microbial killing by secretion of oxidative metab- truding pseudopods (9). Fc␥R-mediated phagocytosis has been shown to have another morphology with protruding pseudopodia *Divison of Infectious Diseases, University of Geneva, Switzerland; †Department for encircling the prey, thus demanding another kind of actin remod- Medical Microbiology, University of Linko¨ping, Sweden; and ‡European Molecular eling (9). Fc␥R-receptor stimulation mediates signaling via im- Biology Laboratory Mouse Biology Program, Monterotondo, Italy mune receptor tyrosine-based activation motifs (ITAM) with the Received for publication August 16, 1999. Accepted for publication June 16, 2000. initiation of a signaling cascade involving phosphorylation of syk, The costs of publication of this article were defrayed in part by the payment of page src-kinases activation of PI3 kinases, rac and phospholipase C, that charges. This article must therefore be hereby marked advertisement in accordance with 18 U.S.C. Section 1734 solely to indicate this fact. stimulate protein kinase C-activation and calcium release from in- 1 This work was supported by Grants from the Swiss National Fund 31-45891.95 and tracellular stores (7–9). 31-50786.97, Swedish Medical Council no 5968, the Swedish Rheumatoid Founda- An actin ring that slowly dissolves after ingestion is completed tion, the Foundation Carlos et Elsie de Reuter, and the Socie´te´ Acade´mique de encircles the phagosome (10). The actin ring may be a barrier, Gene`ve. preventing granules from fusion with the phagosome. Gelsolin 2 Address correspondence and reprint requests to Dr. Lena Serrander, Division of Infectious Diseases, University Hospital of Geneva, CH-1211 Geneva 14, Switzer- translocates to the actin ring and might be important for its re- land. E-mail address: [email protected] moval (P.S. and O.S., unpublished observations). Intracellular 2ϩ 3 Current address: Laboratory Biology of Aging, Department of Geriatrics, University Ca stores and granules also translocate to the phagosome and Hospitals of Geneva, Chemin Petit-Bel-Air 2, CH-1225 Cheˆne-Bourg, Switzerland. phagolysosome fusion takes place (11). In neutrophils, this is a 4 Current address: Immunology Laboratory, Science Faculty, University Nancy 1, calcium-dependent process (12). B. P. 239, F-54506 Vandoeuvre-les-Nancy, France. Extracellular secretion of granule proteins is another neutrophil 5 Abbreviations used in this paper: GsnϪ, Gelsolin-null; E.S., external solution; Fc␥R, Fc␥ receptor; CR, complement receptor; MPO, myeloperoxidase; NBT, nitro- activity, important for killing, but also causing tissue damage. The blue tetrazolium; FRAP, fluorescence recovery after photobleaching. cortical actin network may form a physical barrier that prevents Copyright © 2000 by The American Association of Immunologists 0022-1767/00/$02.00 2452 THE ROLE OF GELSOLIN IN NEUTROPHIL PHAGOCYTOSIS granules from reaching the plasma/phagosomal membrane. The FACS analysis role of gelsolin could be to break up this barrier. It has indeed been Mouse neutrophils (20,000 cells/tube) were pelleted and resuspended in 0.1 shown that actin depolymerization in itself is sufficient to induce ml cold 4% paraformaldehyde. After 30 min, the cells were washed twice secretion in some cell types and that gelsolin can stimulate secre- and incubated for 1 h with 0.1 ml either rat monoclonal anti-mouse tion (13–15). CD16/32 (clone 2.4G2) or anti-mouse CD18 (clone C71/16) diluted 1/100 Ϫ in 0.1% BSA/PBS. After one wash, 0.1 ml of an FITC-labeled secondary We have investigated neutrophils from Gsn mice to study the Ј Ab, goat anti-rat F(ab )2, -fragments diluted 1/200 in 0.1% BSA/PBS sup- role of gelsolin in neutrophil functions. Our results show the fea- plemented with 5% goat IgG, was added. Cells were washed and resus- sibility of using transgenic mice to study the specific role of a pended in 0.2 ml PBS. protein in peripheral blood neutrophils. Gelsolin is the prototype of Cell labeling and fluorescence recovery after photobleaching actin severing and capping proteins. It is thought to play a central (FRAP) role in the actin dynamics of phagocytes. We show here that gel- solin plays an important role in Fc␥R-mediated phagocytosis, but Neutrophils (106/ml) were incubated at 4°C for 1 h with 100 ␮l Krebs- not in several other phagocyte functions.
Load more

Recommended publications
  • The Wiskott-Aldrich Syndrome: the Actin Cytoskeleton and Immune Cell Function
    Disease Markers 29 (2010) 157–175 157 DOI 10.3233/DMA-2010-0735 IOS Press The Wiskott-Aldrich syndrome: The actin cytoskeleton and immune cell function Michael P. Blundella, Austen Wortha,b, Gerben Boumaa and Adrian J. Thrashera,b,∗ aMolecular Immunology Unit, UCL Institute of Child Health, London, UK bDepartment of Immunology, Great Ormond Street Hospital NHS Trust, Great Ormond Street, London, UK Abstract. Wiskott-Aldrich syndrome (WAS) is a rare X-linked recessive primary immunodeficiency characterised by immune dysregulation, microthrombocytopaenia, eczema and lymphoid malignancies. Mutations in the WAS gene can lead to distinct syndrome variations which largely, although not exclusively, depend upon the mutation. Premature termination and deletions abrogate Wiskott-Aldrich syndrome protein (WASp) expression and lead to severe disease (WAS). Missense mutations usually result in reduced protein expression and the phenotypically milder X-linked thrombocytopenia (XLT) or attenuated WAS [1–3]. More recently however novel activating mutations have been described that give rise to X-linked neutropenia (XLN), a third syndrome defined by neutropenia with variable myelodysplasia [4–6]. WASP is key in transducing signals from the cell surface to the actin cytoskeleton, and a lack of WASp results in cytoskeletal defects that compromise multiple aspects of normal cellular activity including proliferation, phagocytosis, immune synapse formation, adhesion and directed migration. Keywords: Wiskott-Aldrich syndrome, actin polymerization, lymphocytes,
    [Show full text]
  • Vinculin Antibody Cat
    Vinculin Antibody Cat. No.: 7809 Vinculin Antibody Figure 2 Western Blot Validation in Human, Mouse and Rat Cell Lines Figure 3 Western Blot Validation in PC3 Cell Lysate Loading: 15 μg of lysates per lane. Antibodies: Vinculin Loading: 15 μg of lysates per lane. Antibodies: Vinculin 7809 (1 μg/mL), 1h incubation at RT in 5% NFDM/TBST. 7809 (Lane 1: 0.5 μg/mL and Lane 2: 1 μg/mL), 1h Secondary: Goat anti-rabbit IgG HRP conjugate at 1:10000 incubation at RT in 5% NFDM/TBST. Secondary: Goat anti- dilution. rabbit IgG HRP conjugate at 1:10000 dilution. Figure 4 Immunocytochemistry Validation of Vinculin in Jurkat Cells Figure 5 Immunofluorescence Validation of Vinculin in Immunocytochemical analysis of Jurkat cells using anti- Jurkat Cells Vinculin antibody (7809) at 5 μg/ml. Cells was fixed with Immunofluorescent analysis of 4% paraformaldehyde- formaldehyde and blocked with 10% serum for 1 h at RT; fixed Jurkat cells labeling Vinculin with 7809 at 20 μg/mL, antigen retrieval was by heat mediation with a citrate followed by goat anti-rabbit IgG secondary antibody at buffer (pH6). Samples were incubated with primary 1/500 dilution (green) and DAPI staining (blue). antibody overnight at 4˚C. A goat anti-rabbit IgG H&L (HRP) at 1/250 was used as secondary. Counter stained with Hematoxylin. Specifications September 23, 2021 1 https://www.prosci-inc.com/vinculin-antibody-7809.html HOST SPECIES: Rabbit SPECIES REACTIVITY: Human, Mouse, Rat HOMOLOGY: Predicted species reactivity based on immunogen sequence: Pig: (100%) Anti-Vinculin antibody (7809) was raised against a peptide corresponding to 23 amino IMMUNOGEN: acids near the center of human Vinculin.
    [Show full text]
  • A Unique and Specific Interaction Between Αt-Catenin and Plakophilin
    2126 Research Article A unique and specific interaction between ␣T-catenin and plakophilin-2 in the area composita, the mixed-type junctional structure of cardiac intercalated discs Steven Goossens1,2,*, Barbara Janssens1,2,*,‡, Stefan Bonné1,2,§, Riet De Rycke1,2, Filip Braet1,2,¶, Jolanda van Hengel1,2 and Frans van Roy1,2,** 1Department for Molecular Biomedical Research, VIB and 2Department of Molecular Biology, Ghent University, B-9052 Ghent, Belgium *These authors contributed equally to this work ‡Present address: Wiley-VCH, Boschstrasse 12, D-69469 Weinheim, Germany §Present address: Diabetes Research Center, Brussels Free University (VUB), Laarbeeklaan 103, B-1090 Brussels, Belgium ¶Present address: Australian Key Centre for Microscopy and Microanalysis, The University of Sydney, NSW 2006, Australia **Author for correspondence (e-mail: [email protected]) Accepted 24 April 2007 Journal of Cell Science 120, 2126-2136 Published by The Company of Biologists 2007 doi:10.1242/jcs.004713 Summary Alpha-catenins play key functional roles in cadherin- desmosomal proteins in the heart localize not only to the catenin cell-cell adhesion complexes. We previously desmosomes in the intercalated discs but also at adhering reported on ␣T-catenin, a novel member of the ␣-catenin junctions with hybrid composition. We found that in the protein family. ␣T-catenin is expressed predominantly in latter junctions, endogenous plakophilin-2 colocalizes with cardiomyocytes, where it colocalizes with ␣E-catenin at the ␣T-catenin. By providing an extra link between the intercalated discs. Whether ␣T- and ␣E-catenin have cadherin-catenin complex and intermediate filaments, the specific or synergistic functions remains unknown. In this binding of ␣T-catenin to plakophilin-2 is proposed to be a study we used the yeast two-hybrid approach to identify means of modulating and strengthening cell-cell adhesion specific functions of ␣T-catenin.
    [Show full text]
  • Calcium Signaling at the Endoplasmic Reticulum Fine-Tuning Stress
    Cell Calcium 70 (2018) 24–31 Contents lists available at ScienceDirect Cell Calcium journal homepage: www.elsevier.com/locate/ceca Review Calcium signaling at the endoplasmic reticulum: fine-tuning stress responses T ⁎ Amado Carreras-Suredaa,b,c, Philippe Pihána,b,c, Claudio Hetza,b,c,d,e, a Center for Geroscience, Brain Health and Metabolism, Faculty of Medicine, University of Chile, Chile b Biomedical Neuroscience Institute, Faculty of Medicine, University of Chile, Santiago, Chile c Program of Cellular and Molecular Biology, Institute of Biomedical Sciences, University of Chile, Santiago, Chile d Buck Institute for Research on Aging, Novato, CA, 94945, USA e Department of Immunology and Infectious Diseases, Harvard School of Public Health, Boston, MA 02115, USA ARTICLE INFO ABSTRACT Keywords: Endoplasmic reticulum (ER) calcium signaling is implicated in a myriad of coordinated cellular processes. The ER homeostasis ER calcium content is tightly regulated as it allows a favorable environment for protein folding, in addition to ER stress operate as a major reservoir for fast and specific release of calcium. Altered ER homeostasis impacts protein Calcium handling mechanisms folding, activating the unfolded protein response (UPR) as a rescue mechanism to restore proteostasis. ER cal- Calcium homeostasis cium release impacts mitochondrial metabolism and also fine-tunes the threshold to undergo apoptosis under Unfolded protein response chronic stress. The global coordination between UPR signaling and energetic demands takes place at mi- Mitochondrial associated membranes Mitochondria biology tochondrial associated membranes (MAMs), specialized subdomains mediating interorganelle communication. Here we discuss current models explaining the functional relationship between ER homeostasis and various cellular responses to coordinate proteostasis and metabolic maintenance.
    [Show full text]
  • The Role of the Interaction of the Vinculin Proline-Rich Linker Region
    ß 2014. Published by The Company of Biologists Ltd | Journal of Cell Science (2014) 127, 1875–1886 doi:10.1242/jcs.133645 RESEARCH ARTICLE The role of the interaction of the vinculin proline-rich linker region with vinexin a in sensing the stiffness of the extracellular matrix Hiroshi Yamashita1,*, Takafumi Ichikawa1,*, Daisuke Matsuyama1, Yasuhisa Kimura1, Kazumitsu Ueda1,2, Susan W. Craig3, Ichiro Harada4 and Noriyuki Kioka1,` ABSTRACT information into biochemical signals. At FAs, transmembrane ECM receptors, integrins, are connected to the ends of actin stress Although extracellular matrix (ECM) stiffness is an important aspect fibers through various cytoplasmic proteins, including talin and of the extracellular microenvironment and is known to direct the vinculin. Cells sense ECM stiffness by ‘pulling’ on the ECM lineage specification of stem cells and affect cancer progression, using actomyosin-generated intracellular tension through FAs and the molecular mechanisms that sense ECM stiffness have not yet adjusting the tension that they exert accordingly (Geiger et al., been elucidated. In this study, we show that the proline-rich linker 2009; Parsons et al., 2010); cells on rigid ECM exert higher (PRL) region of vinculin and the PRL-region-binding protein vinexin intracellular tension than cells on soft ECM. Although some FA are involved in sensing the stiffness of ECM substrates. A rigid proteins have been suggested to engage in sensing intracellular substrate increases the level of cytoskeleton-associated vinculin, tension (del Rio et al., 2009; Friedland et al., 2009; Sawada et al., and the fraction of vinculin stably localizing at focal adhesions (FAs) 2006), the signaling mechanisms by which cells sense ECM is larger on rigid ECM than on soft ECM.
    [Show full text]
  • Calcium Signaling in Aging and Neurodegenerative Diseases 2019
    International Journal of Molecular Sciences Meeting Report Calcium Signaling in Aging and Neurodegenerative Diseases 2019 Luísa Cortes 1,2 , João Malva 2,3,4, Ana Cristina Rego 1,2,3 and Cláudia F. Pereira 1,2,3,* 1 Center for Neuroscience and Cell Biology (CNC), University of Coimbra, Rua Larga, Faculty of Medicine, Polo I, 1st floor, 3004-504 Coimbra, Portugal; [email protected] (L.C.); [email protected] (A.C.R.) 2 CIBB-Center for Innovative Biomedicine and Biotechnology, University of Coimbra, Rua Larga, Faculty of Medicine, Polo I, 1st floor, 3004-504 Coimbra, Portugal; [email protected] 3 Faculty of Medicine, Azinhaga de Santa Comba, Celas, 3000-548 Coimbra, Portugal 4 iCRB- Coimbra Institute for Clinical and Biomedical Research; Azinhaga de Santa Comba, Celas, 3000-548 Coimbra, Portugal * Correspondence: [email protected] Received: 28 December 2019; Accepted: 4 February 2020; Published: 7 February 2020 Abstract: The European Calcium Society (ECS) workshop, which is held every 2 years, is a dedicated meeting of scientists interested in the elucidation of the action of calcium binding, calcium signaling and the study of proteins and organelles, such as mitochondria and endoplasmic reticulum, thereby involved, either in health and disease conditions. The 8th edition of the ECS workshop was organized by a group of researchers from the University of Coimbra, Portugal, in close collaboration with ECS board members. Thanks to the central role of “Calcium Signaling in Aging and Neurodegenerative Disorders”, the ECS 2019 workshop was attended by 62 experts who presented their results in a plenary lecture and five regular symposia, two oral communication sessions and two poster sessions, followed by a hands-on session on calcium imaging.
    [Show full text]
  • The Relationship Between Intermediate Filaments and Microfilaments Before and During the Formation of Desmosomes and Adherens-Ty
    Published May 1, 1987 The Relationship between Intermediate Filaments and Microfilaments before and during the Formation of Desmosomes and Adherens-type Junctions in Mouse Epidermal Keratinocytes Kathleen J. Green, Benjamin Geiger,* Jonathan C. R. Jones, John C. Talian, and Robert D. Goldman Department of Cell Biology and Anatomy, Northwestern University Medical School, Chicago, Illinois 60611; and * Department of Chemical Immunology, The Weizmann Institute of Science, Rehovot, Israel Abstract. Actin, keratin, vinculin and desmoplakin ermost of the concentric MFB. Individual IF often organization were studied in primary mouse keratino- splay out, becoming interwoven into these MFB in the cytes before and during Ca2+-induced cell contact forma- region of cell-substrate contact. In the first 30 min af- tion. Double-label fluorescence shows that in cells cul- ter the Ca 2+ switch, areas of submembranous dense Downloaded from tured in low Ca 2÷ medium, keratin-containing inter- material (identified as adherens junctions), which are mediate filament bundles (IFB) and desmoplakin- associated with the perpendicular MFB, can be seen at containing spots are both concentrated towards the cell newly formed cell-ceU contact sites. By 1-2 h, IFB- center in a region bounded by a series of concentric desmosomal component complexes are aligned with microfilament bundles (MFB). Within 5-30 min after the perpendicular MFB as the complexes become jcb.rupress.org raising Ca 2+ levels, a discontinuous actin/vinculin-rich, redistributed to cell-cell interfaces. Cytochalasin D submembranous zone of fluorescence appears at cell- treatment causes the redistribution of actin into numer- cell interfaces. This zone is usually associated with ous patches; keratin-containing Lr:B undergo a con- short, perpendicular MFB, which become wider and comitant redistribution, forming foci that coincide with longer with time.
    [Show full text]
  • Non-Muscle Myosin 2A (NM2A): Structure, Regulation and Function
    cells Review Non-Muscle Myosin 2A (NM2A): Structure, Regulation and Function Cláudia Brito 1,2 and Sandra Sousa 1,* 1 Group of Cell Biology of Bacterial Infections, i3S-Instituto de Investigação e Inovação em Saúde, IBMC, Universidade do Porto, 4200-135 Porto, Portugal; [email protected] 2 Programa Doutoral em Biologia Molecular e Celular (MCBiology), Instituto de Ciências Biomédicas Abel Salazar, Universidade do Porto, 4099-002 Porto, Portugal * Correspondence: [email protected] Received: 19 May 2020; Accepted: 29 June 2020; Published: 1 July 2020 Abstract: Non-muscle myosin 2A (NM2A) is a motor cytoskeletal enzyme with crucial importance from the early stages of development until adulthood. Due to its capacity to convert chemical energy into force, NM2A powers the contraction of the actomyosin cytoskeleton, required for proper cell division, adhesion and migration, among other cellular functions. Although NM2A has been extensively studied, new findings revealed that a lot remains to be discovered concerning its spatiotemporal regulation in the intracellular environment. In recent years, new functions were attributed to NM2A and its activity was associated to a plethora of illnesses, including neurological disorders and infectious diseases. Here, we provide a concise overview on the current knowledge regarding the structure, the function and the regulation of NM2A. In addition, we recapitulate NM2A-associated diseases and discuss its potential as a therapeutic target. Keywords: non-muscle myosin 2A (NM2A); NM2A activity regulation; NM2A filament assembly; actomyosin cytoskeleton; cell migration; cell adhesion; plasma membrane blebbing 1. Superfamily of Myosins The cell cytoskeleton is an interconnected and dynamic network of filaments essential for intracellular organization and cell shape maintenance.
    [Show full text]
  • Evidence for Rho Kinase Pathway
    Oncogene (2001) 20, 2112 ± 2121 ã 2001 Nature Publishing Group All rights reserved 0950 ± 9232/01 $15.00 www.nature.com/onc Cytoskeletal organization in tropomyosin-mediated reversion of ras-transformation: Evidence for Rho kinase pathway Vanya Shah3, Shantaram Bharadwaj1,2, Kozo Kaibuchi4 and GL Prasad*,1,2 1Department of General Surgery, Wake Forest University School of Medicine, Winston-Salem, North Carolina, NC 27157, USA; 2Department of Cancer Biology, Wake Forest University School of Medicine, Winston-Salem, North Carolina, NC 27157, USA; 3Wistar Institute of Anatomy and Cell Biology, Philadelphia, Pennsylvania, USA; 4Nara Institute of Science and Technology Ikoma, Japan Tropomyosin (TM) family of cytoskeletal proteins is and tropomyosins (TMs) are suppressed to varying implicated in stabilizing actin micro®laments. Many TM degrees in many transformed cells (Ben-Ze'ev, 1997). isoforms, including tropomyosin-1 (TM1), are down- Furthermore, restoration of these proteins inhibits the regulated in transformed cells. Previously we demon- malignant phenotype of many dierent experimentally strated that TM1 is a suppressor of the malignant transformed cell lines, underscoring the pivotal role of transformation, and that TM1 reorganizes micro®la- cytoskeletal organization in maintaining a normal ments in the transformed cells. To investigate how TM1 phenotype (Ayscough, 1998; Janmey and Chaponnier, induces micro®lament organization in transformed cells, 1995). Our laboratory has been interested in under- we utilized ras-transformed NIH3T3 (DT) cells, and standing the role of cytoskeletal proteins, in particular those transduced to express TM1, and/or TM2. that of tropomyosins, in malignant transformation. Enhanced expression of TM1 alone, but not TM2, Tropomyosin (TM) family comprises of 5 ± 7 results in re-emergence of micro®laments; TM1, together dierent closely related isoforms, whose expression is with TM2 remarkably improves micro®lament architec- altered in many transformed cells (Lin et al., 1997; ture.
    [Show full text]
  • Snapshot: Actin Regulators II Anosha D
    SnapShot: Actin Regulators II Anosha D. Siripala and Matthew D. Welch Department of Molecular and Cell Biology, University of California, Berkeley, CA 94720, USA Representative Proteins Protein Family H. sapiens D. melanogaster C. elegans A. thaliana S. cerevisiae Endocytosis and Exocytosis ABP1/drebrin mABP1, drebrin, drebrin- †Q95RN0 †Q9XUT0 Abp1 like EPS15 EPS15 Eps-15 EHS-1 †Q56WL2 Pan1 HIP1R HIP1R †Q8MQK1 †O62142 Sla2 Synapsin synapsin Ia, Ib, IIa, IIb, III Synapsin SNN-1 Plasma Membrane Association Anillin anillin Scraps ANI-1, 2, 3 Annexins annexin A1–11, 13 (actin Annexin B9-11 NEX-1–4 ANN1-8 binding: 1, 2, 6) ERM proteins ezrin, radixin, moesin DMoesin ERM-1 MARCKS MARCKS, MRP/ Akap200 MACMARCKS/F52 Merlin *merlin/NF2 Merlin NFM-1 Protein 4.1 4.1R, G, N, B Coracle Spectrin α-spectrin (1–2), β-spectrin α-spectrin, β-spectrin, β heavy- SPC-1 (α-spectrin), UNC-70 (1–4), β heavy-spectrin/ spectrin/Karst (β-spectrin), SMA-1 (β heavy- karst spectrin) Identifi ed Cellular Role: X Membrane traffi cking and phagocytosis Cell-Cell Junctions X Cytokinesis α-catenin α-catenin 1–3 α-catenin HMP-1 X Cell surface organization and dynamics X Cell adhesion Afadin afadin/AF6 Canoe AFD-1 X Multiple functions ZO-1 ZO-1, ZO-2, ZO-3 ZO-1/Polychaetoid †Q56VX4 X Other/unknown Cell-Extracellular Matrix Junctions †UNIPROT database accession number *Mutation linked to human disease Dystrophin/utrophin *dystrophin, utrophin/ Dystrophin DYS-1 DRP1, DRP2 LASP LASP-1, LASP-2, LIM- Lasp †P34416 nebulette Palladin palladin Parvin α-, β-, χ-parvin †Q9VWD0 PAT-6
    [Show full text]
  • Protein Network Analyses of Pulmonary Endothelial Cells In
    www.nature.com/scientificreports OPEN Protein network analyses of pulmonary endothelial cells in chronic thromboembolic pulmonary hypertension Sarath Babu Nukala1,8,9*, Olga Tura‑Ceide3,4,5,9, Giancarlo Aldini1, Valérie F. E. D. Smolders2,3, Isabel Blanco3,4, Victor I. Peinado3,4, Manuel Castell6, Joan Albert Barber3,4, Alessandra Altomare1, Giovanna Baron1, Marina Carini1, Marta Cascante2,7,9 & Alfonsina D’Amato1,9* Chronic thromboembolic pulmonary hypertension (CTEPH) is a vascular disease characterized by the presence of organized thromboembolic material in pulmonary arteries leading to increased vascular resistance, heart failure and death. Dysfunction of endothelial cells is involved in CTEPH. The present study describes for the frst time the molecular processes underlying endothelial dysfunction in the development of the CTEPH. The advanced analytical approach and the protein network analyses of patient derived CTEPH endothelial cells allowed the quantitation of 3258 proteins. The 673 diferentially regulated proteins were associated with functional and disease protein network modules. The protein network analyses resulted in the characterization of dysregulated pathways associated with endothelial dysfunction, such as mitochondrial dysfunction, oxidative phosphorylation, sirtuin signaling, infammatory response, oxidative stress and fatty acid metabolism related pathways. In addition, the quantifcation of advanced oxidation protein products, total protein carbonyl content, and intracellular reactive oxygen species resulted increased
    [Show full text]
  • Neuronal Calcium Sensor-1 Enhancement of Insp3 Receptor Activity Is Inhibited by Therapeutic Levels of Lithium
    Neuronal calcium sensor-1 enhancement of InsP3 receptor activity is inhibited by therapeutic levels of lithium Christina Schlecker, … , Klara Szigeti-Buck, Barbara E. Ehrlich J Clin Invest. 2006;116(6):1668-1674. https://doi.org/10.1172/JCI22466. Research Article Neuroscience Regulation and dysregulation of intracellular calcium (Ca2+) signaling via the inositol 1,4,5-trisphosphate receptor (InsP3R) has been linked to many cellular processes and pathological conditions. In the present study, addition of neuronal calcium sensor-1 (NCS-1), a high-affinity, low-capacity, calcium-binding protein, to purified InsP3R type 1 (InsP3R1) increased the channel activity in both a calcium-dependent and -independent manner. In intact cells, enhanced expression of NCS-1 resulted in increased intracellular calcium release upon stimulation of the phosphoinositide signaling pathway. To determine whether InsP3R1/NCS-1 interaction could be functionally relevant in bipolar disorders, conditions in which NCS-1 is highly expressed, we tested the effect of lithium, a salt widely used for treatment of bipolar disorders. Lithium inhibited the enhancing effect of NCS-1 on InsP3R1 function, suggesting that InsP3R1/NCS-1 interaction is an essential component of the pathomechanism of bipolar disorder. Find the latest version: https://jci.me/22466/pdf Research article Neuronal calcium sensor-1 enhancement of InsP3 receptor activity is inhibited by therapeutic levels of lithium Christina Schlecker,1,2,3 Wolfgang Boehmerle,1,4 Andreas Jeromin,5 Brenda DeGray,1 Anurag Varshney,1 Yogendra Sharma,6 Klara Szigeti-Buck,7 and Barbara E. Ehrlich1,3 1Department of Pharmacology, Yale University, New Haven, Connecticut, USA. 2Department of Neuroscience, University of Magdeburg, Magdeburg, Germany.
    [Show full text]