DOCSLIB.ORG

Differential Effects of Desmoglein 1 and Desmoglein 3 on Desmosome

Total Page：16

File Type：pdf, Size：1020Kb

Download full-text PDF Read full-text
Differential Effects of Desmoglein 1 and Desmoglein 3 on Desmosome View metadata, citation and similar papers at core.ac.uk brought to you by CORE providedORIGINAL by Elsevier - ARTICLE Publisher Connector See related Commentary on page 1215 Di¡erential E¡ects of Desmoglein 1 and Desmoglein 3 on Desmosome Formation Yasushi Hanakawa, Masayuki Amagai,n Yuji Shirakata, Yoko Yahata, Sho Tokumaru, Kenshi Yamasaki, Mikiko Tohyama, Koji Sayama, and Koji Hashimoto Department of Dermatology, School of Medicine, Ehime University, Ehime, Japan; nDepartment of Dermatology, School of Medicine, Keio University, Tokyo, Japan The desmoglein plays an important part in the forma- desmoglein 3DEC. Therefore, we conclude that the tion of desmosomes. We constructed recombinant ade- dominant-negative e¡ect of desmoglein 1DEC is not noviruses containing desmoglein 1 and desmoglein 3 simply due to plakoglobin sequestration. On the other derivatives partly lacking the extracellular domain (des- hand, during low-level expression of full-length desmo- moglein 1DEC and desmoglein 3DEC, respectively), and glein 3 and desmoglein 1, they both colocalized with full-length desmoglein 1 and desmoglein 3 and studied desmoplakin. During high-level expression, however, the involvement of desmoglein 1 and desmoglein 3 in keratin insertion at cell^cell contact sites was inhibited desmosome formation. During low-level expression in desmoglein 1 but not in desmoglein 3, and desmopla- of desmoglein 3DEC in transduced HaCaTcells, keratin kin was stained at cell^cell contact sites in desmoglein 3 insertion at cell^cell contact sites was only partially but not in desmoglein 1. These data suggest desmoglein inhibited and desmoplakin was partially stained at 1 and desmoglein 3 expressed at low level were incorpo- cell^cell contact sites. Low-level expression of desmo- rated into desmosome but at high-level expression, des- glein 1DEC, however, resulted in complete inhibition moglein 1 disrupted desmosomes but desmoglein 3 did of keratin insertion at the cell^cell contact sites, and not. Our ¢ndings provide biologic evidence that des- desmoplakin was stained in perinuclear dots. These moglein 1 and desmoglein 3 play a di¡erent functional results indicate the dominant-negative e¡ect of desmo- role in cell^cell adhesion of keratinocytes. Key words: glein 1DEC on desmosome formation was stronger than desmosomes/desmoglein 1/desmoglein 3. J Invest Dermatol that of desmoglein 3DEC. Desmoglein 1DEC coprecipi- 119:1231 ^1236, 2002 tated plakoglobin to approximately the same extent as esmosomes are cell^cell adhesion complexes that is a disease caused by autoantibodies directed against Dsg3 in provide mechanical integrity to keratinocytes by which skin lesion biopsies exhibit suprabasilar acantholysis (Udey linking to keratin intermediate ¢laments. Desmo- and Stanley, 1999). On the other hand, pemphigus foliaceus is somes are composed of two major transmembrane caused by autoantibodies directed against Dsg1 and in this case, proteins, desmoglein (Dsg) and desmocollin (Ama- lesion biopsies exhibit subcorneal acantholysis. Dsg3 knockout Dgai, 1996a; Kowalczyck et al, 1999; Green and Gaudry, 2000). In mice phenotypically mimicked pemphigus vulgaris patients humans, three desmoglein isoforms have been identi¢ed: Dsg1, (Koch et al, 1997) in that they displayed oral erosions and loss of Dsg2, and Dsg3. They are encoded by individual genes and dif- intercellular adhesion of suprabasal layers of the mucosal epithe- ferentially distributed in tissue. Dsg2 is expressed in all desmo- lium and epidermis. some-containing tissues, including simple epithelium and Several studies have suggested that both the extracellular and myocardium. In contrast, Dsg1 and Dsg3 are expressed cytoplasmic domains of desmogleins are critical for normal des- in strati¢ed squamous epithelia. Dsg3 is found in the basal and mosome formation. N-terminally truncated Dsg3 caused domi- suprabasal layers of stratifying epithelia, whereas Dsg1 is nant-negative e¡ects on desmosome formation in HaCaT cells dominantly expressed in the di¡erentiated upper layers of (Hanakawa et al, 2000). Expression of chimeric molecules con- epithelia (Arnemann et al, 1993; Shimizu et al,1995;Amagaiet al, taining the transmembrane domain of connexin and cytoplasmic 1996b). domain of Dsg1 disrupted desmosomes in A431 cells (Troyanovs- Desmogleins play important parts in the formation and main- ky et al, 1993). Similarly, a chimeric molecule containing the ex- tenance of desmosomes. Autoantibodies against desmogleins lead tracellular domain of E-cadherin and cytoplasmic domain of to impairment of epidermal tissue integrity. Pemphigus vulgaris Dsg1 (Ecad-Dsg1) disrupted desmosomes in A431 cells (Norvel and Green, 1998). In contrast, a recent report indicated that a chi- meric molecule of the extracellular domain of E-cadherin and Manuscript received January 10, 2002; revised June 24, 2002; accepted for cytoplasmic domain of Dsg3 (Ecad-Dsg3) was incorporated into publication August 29, 2002 Reprint requests to: Yasushi Hanakawa, MD, Ehime University School desmosomes of A431 cells (Andl and Stanley, 2001). This di¡er- of Medicine, Department of Dermatology, Shitukawa, Shigenobu, Onsen- ence between Ecad-Dsg1 and Ecad-Dsg3 indicates functional dif- gun, Ehime 791-0295, Japan. Email: [email protected] ferences of cytoplasmic domains of Dsg1 and Dsg3. In addition, Abbreviations: Dsg, desmoglein; MOI, multiplicity of infection. full-length Dsg1 disrupted desmosome when the expression level 0022-202X/02/$15.00 Copyright r 2002 by The Society for Investigative Dermatology, Inc. 1231 1232 HANAKAWA ET AL THE JOURNAL OF INVESTIGATIVE DERMATOLOGY was high; however, full-length Dsg1 was incorporated into des- (murine anti-desmoplakin; PROGEN); PG5.1 (murine anti-plakoglobin; mosome when the expression level was low in A431 cells (Norvel PROGEN); PP1-5C2 (murine anti-plakophilin 1; PROGEN); anti- and Green, 1998). These data suggest that Dsg1 and Dsg3 expres- plakophilin 2 (murine; PROGEN); anti-involucrin (Abcam, Cambridge, sion have di¡erent e¡ects on desmosome formation and integrity. U.K.); and 6H6 (murine anti-Dsg3). The following rabbit anti-sera were used: Z622 (anti-pan-keratin; DAKO, Copenhagen, Denmark); anti-myc To characterize further and de¢ne these di¡erences, we con- (a kind gift from Dr John Stanley, University of Pennsylvania, PA); anti- structed full-length and N-terminally truncated mutants of keratin 1 (CONVENCE, Richmond, CA); anti-loricrin (CONVENCE); Dsg1 and Dsg3, and introduced them into cultured keratinocytes and anti-FLAG (Abcam). Fluorescein isothiocyanate-conjugated goat anti- using an adenovirus vector. Taking advantage of the adenovirus mouse or goat anti-rabbit antibodies (Kirkegaard and Perry Laboratories, vector to control expression level by changing multiplicity of Gaithersburg, MD) and rhodamine-conjugated goat anti-rabbit antibody infection (MOI), we evaluated the e¡ects of expression levels of (BioSource, Camarillo, CA) were used. these isoforms on the integrity of a desmosome^keratin struc- tured complex. Immunohistochemistry Cells grown on Laboratory-Tech 4-well culture slides (Nalge-Nunc, Napierville, IL) were ¢xed with methanol at ^201C for 20 min and permeabilized with 0.05% Triton X-100 in Tris- 2 þ bu¡ered saline containing 1 mM CaCl2 (TBS-Ca ) at room temperature MATERIALS AND METHODS for 5 min. After incubation with 1% bovine serum albumin in TBS-Ca2 þ for 20 min at room temperature, the cells were incubated with various Cell culture The human embryonic kidney cell line 293 was obtained antibodies at the appropriate dilution in 1% bovine serum albumin in from American TypeTissue Culture (ATCC; Rockville, MD). The human TBS-Ca2 þ for 1 h at room temperature. The samples were further naturally immortalized keratinocyte HaCaTcell line was a kind gift from incubated with £uorescein isothiocyanate- or rhodamine-conjugated Dr Norbert Fusenig (German Cancer Research Center, Heidelberg, secondary antibodies at the appropriate dilution in 1% bovine serum Germany). These cells were cultured in Dulbecco modi¢ed Eagle’s albumin in TBS-Ca2 þ for 1 h at room temperature. Stained cells were medium supplemented with 10% fetal bovine serum. examined using a confocal laser microscope (Zeiss, Oberkachen, Germany). Plasmid construction The full-length cDNA encoding human Dsg1 was a kind gift from Dr Kathleen Green (North-western University, Immunoblotting and immunoprecipitation For immunoblotting, Chicago, IL). The construction of the Dsg3 mutant, in which a large part cells were lyzed in sodium dodecyl sulfate sample bu¡er (62.5 mM Tris^ of the extracellular domain was deleted and a seven c-myc tag was inserted HCl, pH 7.5, 1% sodium dodecyl sulfate, 0.0025% bromophenol blue, £anking the C-terminal end, has been described previously (Hanakawa 10% glycerol, 2.5% 2-mercaptoethanol) and subjected to immunoblotting et al, 2000). A mutant of Dsg1 (Dsg1DEC), with a large part of the using enhanced chemi£uorescence according to the manufacturer’s extracellular domain deleted and with a seven c-myc tag £anking the protocol (Amersham Pharmacia Biotech, Uppsala, Sweden). For soluble C-terminal end, was also constructed. Dsg1 DNA fragments (nucleotides and insoluble fractionation, the cells were scraped in 1% Nonidet P-40 13^402 and 1726^3360) were generated by polymerase chain reaction and 1% Trion-X-100 in TBS-Ca2 þ on ice. Following centrifugation at using primers DG1F150 (50 -TTTTAGGGTGGGGATCCAGAC-30), DG1F130 1 0 0 0 15,000 r.p.m. (30,000g) for 10 min at 4 C, the supernatant was collected
Load more

Recommended publications
  • ARVC-Variants.Pdf
    Updated gene list responsible for ARVC/D pathology Subtype Gene Location Reference ARVC1 TGFB3 14q24.3 Beffagna G, Occhi G, Nava A, et al. Regulatory mutations in transforming growth factor-beta3 gene cause arrhythmogenic right ventricular cardiomyopathy type 1. Cardiovasc Res. 2005;65:366–73. ARVC2 RYR2 1q43 Tiso N, Stephan DA, Nava A, et al. Identification of mutations in the cardiac ryanodine receptor gene in families affected with arrhythmogenic right ventricular cardiomyopathy type 2 (ARVD2). Hum Mol Genet. 2001;10:189–94. ARVC3 Unknown 14q12-q22 Severini GM, Krajinovic M, Pinamonti B, et al. A new locus for arrhythmogenic right ventricular dysplasia on the long arm of chromosome 14. Genomics. 1996;31:193–200. ARVC4 TTN 2q32.1-q32.3 Taylor M, Graw S, Sinagra G, et al. Genetic variation in titin in arrhythmogenic right ventricular cardiomyopathy-overlap syndromes. Circulation. 2011;124:876–85. ARVC5 TMEM43 3p25.1 Merner ND, Hodgkinson KA, Haywood AF, et al. Arrhythmogenic right ventricular cardiomyopathy type 5 is a fully penetrant, lethal arrhythmic disorder caused by a missense mutation in the TMEM43 gene. Am J Hum Genet. 2008;82:809–21. ARVC6 Unknown 10p14-p12 Li D, Ahmad F, Gardner MJ, et al. The locus of a novel gene responsible for arrhythmogenic right- ventricular dysplasia characterized by early onset and high penetrance maps to chromosome 10p12-p14. Am J Hum Genet. 2000;66:148–56. ARVC7 DES 2q35 Klauke B, Kossmann S, Gaertner A, et al. De novo desmin-mutation N116S is associated with arrhythmogenic right ventricular cardiomyopathy. Hum Mol Genet. 2010;19:4595–607.
    [Show full text]
  • Vocabulario De Morfoloxía, Anatomía E Citoloxía Veterinaria
    Vocabulario de Morfoloxía, anatomía e citoloxía veterinaria (galego-español-inglés) Servizo de Normalización Lingüística Universidade de Santiago de Compostela COLECCIÓN VOCABULARIOS TEMÁTICOS N.º 4 SERVIZO DE NORMALIZACIÓN LINGÜÍSTICA Vocabulario de Morfoloxía, anatomía e citoloxía veterinaria (galego-español-inglés) 2008 UNIVERSIDADE DE SANTIAGO DE COMPOSTELA VOCABULARIO de morfoloxía, anatomía e citoloxía veterinaria : (galego-español- inglés) / coordinador Xusto A. Rodríguez Río, Servizo de Normalización Lingüística ; autores Matilde Lombardero Fernández ... [et al.]. – Santiago de Compostela : Universidade de Santiago de Compostela, Servizo de Publicacións e Intercambio Científico, 2008. – 369 p. ; 21 cm. – (Vocabularios temáticos ; 4). - D.L. C 2458-2008. – ISBN 978-84-9887-018-3 1.Medicina �������������������������������������������������������������������������veterinaria-Diccionarios�������������������������������������������������. 2.Galego (Lingua)-Glosarios, vocabularios, etc. políglotas. I.Lombardero Fernández, Matilde. II.Rodríguez Rio, Xusto A. coord. III. Universidade de Santiago de Compostela. Servizo de Normalización Lingüística, coord. IV.Universidade de Santiago de Compostela. Servizo de Publicacións e Intercambio Científico, ed. V.Serie. 591.4(038)=699=60=20 Coordinador Xusto A. Rodríguez Río (Área de Terminoloxía. Servizo de Normalización Lingüística. Universidade de Santiago de Compostela) Autoras/res Matilde Lombardero Fernández (doutora en Veterinaria e profesora do Departamento de Anatomía e Produción Animal.
    [Show full text]
  • Plakoglobin Is Required for Effective Intermediate Filament Anchorage to Desmosomes Devrim Acehan1, Christopher Petzold1, Iwona Gumper2, David D
    ORIGINAL ARTICLE Plakoglobin Is Required for Effective Intermediate Filament Anchorage to Desmosomes Devrim Acehan1, Christopher Petzold1, Iwona Gumper2, David D. Sabatini2, Eliane J. Mu¨ller3, Pamela Cowin2,4 and David L. Stokes1,2,5 Desmosomes are adhesive junctions that provide mechanical coupling between cells. Plakoglobin (PG) is a major component of the intracellular plaque that serves to connect transmembrane elements to the cytoskeleton. We have used electron tomography and immunolabeling to investigate the consequences of PG knockout on the molecular architecture of the intracellular plaque in cultured keratinocytes. Although knockout keratinocytes form substantial numbers of desmosome-like junctions and have a relatively normal intercellular distribution of desmosomal cadherins, their cytoplasmic plaques are sparse and anchoring of intermediate filaments is defective. In the knockout, b-catenin appears to substitute for PG in the clustering of cadherins, but is unable to recruit normal levels of plakophilin-1 and desmoplakin to the plaque. By comparing tomograms of wild type and knockout desmosomes, we have assigned particular densities to desmoplakin and described their interaction with intermediate filaments. Desmoplakin molecules are more extended in wild type than knockout desmosomes, as if intermediate filament connections produced tension within the plaque. On the basis of our observations, we propose a particular assembly sequence, beginning with cadherin clustering within the plasma membrane, followed by recruitment of plakophilin and desmoplakin to the plaque, and ending with anchoring of intermediate filaments, which represents the key to adhesive strength. Journal of Investigative Dermatology (2008) 128, 2665–2675; doi:10.1038/jid.2008.141; published online 22 May 2008 INTRODUCTION dense plaque that is further from the membrane and that Desmosomes are large macromolecular complexes that mediates the binding of intermediate filaments.
    [Show full text]
  • Supplementary Table 1: Adhesion Genes Data Set
    Supplementary Table 1: Adhesion genes data set PROBE Entrez Gene ID Celera Gene ID Gene_Symbol Gene_Name 160832 1 hCG201364.3 A1BG alpha-1-B glycoprotein 223658 1 hCG201364.3 A1BG alpha-1-B glycoprotein 212988 102 hCG40040.3 ADAM10 ADAM metallopeptidase domain 10 133411 4185 hCG28232.2 ADAM11 ADAM metallopeptidase domain 11 110695 8038 hCG40937.4 ADAM12 ADAM metallopeptidase domain 12 (meltrin alpha) 195222 8038 hCG40937.4 ADAM12 ADAM metallopeptidase domain 12 (meltrin alpha) 165344 8751 hCG20021.3 ADAM15 ADAM metallopeptidase domain 15 (metargidin) 189065 6868 null ADAM17 ADAM metallopeptidase domain 17 (tumor necrosis factor, alpha, converting enzyme) 108119 8728 hCG15398.4 ADAM19 ADAM metallopeptidase domain 19 (meltrin beta) 117763 8748 hCG20675.3 ADAM20 ADAM metallopeptidase domain 20 126448 8747 hCG1785634.2 ADAM21 ADAM metallopeptidase domain 21 208981 8747 hCG1785634.2|hCG2042897 ADAM21 ADAM metallopeptidase domain 21 180903 53616 hCG17212.4 ADAM22 ADAM metallopeptidase domain 22 177272 8745 hCG1811623.1 ADAM23 ADAM metallopeptidase domain 23 102384 10863 hCG1818505.1 ADAM28 ADAM metallopeptidase domain 28 119968 11086 hCG1786734.2 ADAM29 ADAM metallopeptidase domain 29 205542 11085 hCG1997196.1 ADAM30 ADAM metallopeptidase domain 30 148417 80332 hCG39255.4 ADAM33 ADAM metallopeptidase domain 33 140492 8756 hCG1789002.2 ADAM7 ADAM metallopeptidase domain 7 122603 101 hCG1816947.1 ADAM8 ADAM metallopeptidase domain 8 183965 8754 hCG1996391 ADAM9 ADAM metallopeptidase domain 9 (meltrin gamma) 129974 27299 hCG15447.3 ADAMDEC1 ADAM-like,
    [Show full text]
  • Cell Biology of Tight Junction Barrier Regulation and Mucosal Disease
    Downloaded from http://cshperspectives.cshlp.org/ on October 1, 2021 - Published by Cold Spring Harbor Laboratory Press Cell Biology of Tight Junction Barrier Regulation and Mucosal Disease Aaron Buckley and Jerrold R. Turner Departments of Pathology and Medicine (Gastroenterology), Brigham and Women’s Hospital and Harvard Medical School, Boston, Massachusetts 02115 Correspondence: [email protected] Mucosal surfaces are lined by epithelial cells. In the intestine, the epithelium establishes a selectively permeable barrier that supports nutrient absorption and waste secretion while preventing intrusion by luminal materials. Intestinal epithelia therefore play a central role in regulating interactions between the mucosal immune system and luminal contents, which include dietary antigens, a diverse intestinal microbiome, and pathogens. The paracellular space is sealed by the tight junction, which is maintained by a complex network of protein interactions. Tight junction dysfunction has been linked to a variety of local and systemic diseases. Two molecularly and biophysically distinct pathways across the intestinal tight junc- tion are selectively and differentially regulated by inflammatory stimuli. This review discusses the mechanisms underlying these events, their impact on disease, and the potential of using these as paradigms for development of tight junction-targeted therapeutic interventions. ucosal surfaces and the epithelial cells that adherens). The tight junction is a selectively Mline them are present at sites where tissues permeable barrier that generally represents the interface directly with the external environment rate-limiting step of paracellular transport. The or internal compartments that are contiguous adherens junction and desmosome provide es- with the external environment. Examples in- sential adhesive and mechanical properties that clude the gastrointestinal tract, the pulmonary contribute to barrier function but do not seal tree, and the genitourinary tract.
    [Show full text]
  • Hemidesmosomes Show Abnormal Association with the Keratin Filament Network in Junctional Forms of Epidermolysis Bullosa
    View metadata, citation and similar papers at core.ac.uk brought to you by CORE provided by Elsevier - Publisher Connector Hemidesmosomes Show Abnormal Association with the Keratin Filament Network in Junctional Forms of Epidermolysis Bullosa James R. McMillan, John A. McGrath, Michael J. Tidman,* and Robin A. J. Eady Department of Cell Pathology, St John’s Institute of Dermatology, UMDS, St Thomas’s Hospital, London, U.K.; *Department of Dermatology, Royal Infirmary of Edinburgh, Edinburgh, U.K. Junctional epidermolysis bullosa is a group of hereditary respectively. In junctional epidermolysis bullosa with bullous disorders resulting from defects in several hemi- pyloric atresia (α6β4 abnormalities, n J 3) the values desmosome-anchoring filament components. Because were also reduced [41.8% K 7.0 (p < 0.001) and 44.5% hemidesmosomes are involved not only in keratinocyte- K 5.7 (p < 0.001), respectively]. In the non-Herlitz group extracellular matrix adherence, but also in normal (laminin 5 mutations, n J 3) the counts were 66.7% K anchorage of keratin intermediate filaments to the basal 7.1 (p > 0.05) and 70.5% K 8.5 (p < 0.05), and in keratinocyte membrane, we questioned whether this skin from patients with bullous pemphigoid antigen 2 intracellular function of hemidesmosomes was also per- mutations (n J 3) the counts were 54.3% K 13.8 (p < 0.01) turbed in junctional epidermolysis bullosa. We used and 57.1% K 13.9 (p < 0.01). In epidermolysis bullosa quantitative electron microscopic methods to assess cer- simplex associated with plectin mutations the values were tain morphologic features of hemidesmosome–keratin 31.9% K 8.9 (p < 0.001) for keratin intermediate filaments intermediate filaments interactions in skin from normal association and 39.9% K 7.1 (p < 0.001) for inner plaques.
    [Show full text]
  • Nomina Histologica Veterinaria, First Edition
    NOMINA HISTOLOGICA VETERINARIA Submitted by the International Committee on Veterinary Histological Nomenclature (ICVHN) to the World Association of Veterinary Anatomists Published on the website of the World Association of Veterinary Anatomists www.wava-amav.org 2017 CONTENTS Introduction i Principles of term construction in N.H.V. iii Cytologia – Cytology 1 Textus epithelialis – Epithelial tissue 10 Textus connectivus – Connective tissue 13 Sanguis et Lympha – Blood and Lymph 17 Textus muscularis – Muscle tissue 19 Textus nervosus – Nerve tissue 20 Splanchnologia – Viscera 23 Systema digestorium – Digestive system 24 Systema respiratorium – Respiratory system 32 Systema urinarium – Urinary system 35 Organa genitalia masculina – Male genital system 38 Organa genitalia feminina – Female genital system 42 Systema endocrinum – Endocrine system 45 Systema cardiovasculare et lymphaticum [Angiologia] – Cardiovascular and lymphatic system 47 Systema nervosum – Nervous system 52 Receptores sensorii et Organa sensuum – Sensory receptors and Sense organs 58 Integumentum – Integument 64 INTRODUCTION The preparations leading to the publication of the present first edition of the Nomina Histologica Veterinaria has a long history spanning more than 50 years. Under the auspices of the World Association of Veterinary Anatomists (W.A.V.A.), the International Committee on Veterinary Anatomical Nomenclature (I.C.V.A.N.) appointed in Giessen, 1965, a Subcommittee on Histology and Embryology which started a working relation with the Subcommittee on Histology of the former International Anatomical Nomenclature Committee. In Mexico City, 1971, this Subcommittee presented a document entitled Nomina Histologica Veterinaria: A Working Draft as a basis for the continued work of the newly-appointed Subcommittee on Histological Nomenclature. This resulted in the editing of the Nomina Histologica Veterinaria: A Working Draft II (Toulouse, 1974), followed by preparations for publication of a Nomina Histologica Veterinaria.
    [Show full text]
  • Molecular Organization of the Desmosome As Revealed by Direct Stochastic Optical Reconstruction Microscopy Sara N
    © 2016. Published by The Company of Biologists Ltd | Journal of Cell Science (2016) 129, 2897-2904 doi:10.1242/jcs.185785 SHORT REPORT Molecular organization of the desmosome as revealed by direct stochastic optical reconstruction microscopy Sara N. Stahley1, Emily I. Bartle1, Claire E. Atkinson2, Andrew P. Kowalczyk1,3 and Alexa L. Mattheyses1,* ABSTRACT plakoglobin and plakophilin, and the plakin family member Desmosomes are macromolecular junctions responsible for providing desmoplakin contribute to the intracellular plaque (Fig. 1A). strong cell–cell adhesion. Because of their size and molecular Plaque ultrastructure is characterized by two electron-dense complexity, the precise ultrastructural organization of desmosomes regions: the plasma-membrane-proximal outer dense plaque and is challenging to study. Here, we used direct stochastic optical the inner dense plaque (Desai et al., 2009; Farquhar and Palade, reconstruction microscopy (dSTORM) to resolve individual plaque 1963; Stokes, 2007). The cadherin cytoplasmic tails bind to proteins pairs for inner and outer dense plaque proteins. Analysis methods in the outer dense plaque whereas the C-terminus of desmoplakin based on desmosomal mirror symmetry were developed to measure binds to intermediate filaments in the inner dense plaque. This plaque-to-plaque distances and create an integrated map. We tethers the desmosome to the intermediate filament cytoskeleton, quantified the organization of desmoglein 3, plakoglobin and establishing an integrated adhesive network (Bornslaeger et al., desmoplakin (N-terminal, rod and C-terminal domains) in primary 1996; Harmon and Green, 2013). human keratinocytes. Longer desmosome lengths correlated with Many desmosomal protein interactions have been characterized increasing plaque-to-plaque distance, suggesting that desmoplakin is by biochemical studies (Bass-Zubek and Green, 2007; Green and arranged with its long axis at an angle within the plaque.
    [Show full text]
  • Adherens Junctions, Desmosomes and Tight Junctions in Epidermal Barrier Function Johanna M
    14 The Open Dermatology Journal, 2010, 4, 14-20 Open Access Adherens Junctions, Desmosomes and Tight Junctions in Epidermal Barrier Function Johanna M. Brandner1,§, Marek Haftek*,2,§ and Carien M. Niessen3,§ 1Department of Dermatology and Venerology, University Hospital Hamburg-Eppendorf, Hamburg, Germany 2University of Lyon, EA4169 Normal and Pathological Functions of Skin Barrier, E. Herriot Hospital, Lyon, France 3Department of Dermatology, Center for Molecular Medicine, Cologne Excellence Cluster on Cellular Stress Responses in Aging-Associated Diseases (CECAD), University of Cologne, Germany Abstract: The skin is an indispensable barrier which protects the body from the uncontrolled loss of water and solutes as well as from chemical and physical assaults and the invasion of pathogens. In recent years several studies have suggested an important role of intercellular junctions for the barrier function of the epidermis. In this review we summarize our knowledge of the impact of adherens junctions, (corneo)-desmosomes and tight junctions on barrier function of the skin. Keywords: Cadherins, catenins, claudins, cell polarity, stratum corneum, skin diseases. INTRODUCTION ADHERENS JUNCTIONS The stratifying epidermis of the skin physically separates Adherens junctions are intercellular structures that couple the organism from its environment and serves as its first line intercellular adhesion to the cytoskeleton thereby creating a of structural and functional defense against dehydration, transcellular network that coordinate the behavior of a chemical substances, physical insults and micro-organisms. population of cells. Adherens junctions are dynamic entities The living cell layers of the epidermis are crucial in the and also function as signal platforms that regulate formation and maintenance of the barrier on two different cytoskeletal dynamics and cell polarity.
    [Show full text]
  • Cell Adhesion Molecules in Normal Skin and Melanoma
    biomolecules Review Cell Adhesion Molecules in Normal Skin and Melanoma Cian D’Arcy and Christina Kiel * Systems Biology Ireland & UCD Charles Institute of Dermatology, School of Medicine, University College Dublin, D04 V1W8 Dublin, Ireland; [email protected] * Correspondence: [email protected]; Tel.: +353-1-716-6344 Abstract: Cell adhesion molecules (CAMs) of the cadherin, integrin, immunoglobulin, and selectin protein families are indispensable for the formation and maintenance of multicellular tissues, espe- cially epithelia. In the epidermis, they are involved in cell–cell contacts and in cellular interactions with the extracellular matrix (ECM), thereby contributing to the structural integrity and barrier for- mation of the skin. Bulk and single cell RNA sequencing data show that >170 CAMs are expressed in the healthy human skin, with high expression levels in melanocytes, keratinocytes, endothelial, and smooth muscle cells. Alterations in expression levels of CAMs are involved in melanoma propagation, interaction with the microenvironment, and metastasis. Recent mechanistic analyses together with protein and gene expression data provide a better picture of the role of CAMs in the context of skin physiology and melanoma. Here, we review progress in the field and discuss molecular mechanisms in light of gene expression profiles, including recent single cell RNA expression information. We highlight key adhesion molecules in melanoma, which can guide the identification of pathways and Citation: D’Arcy, C.; Kiel, C. Cell strategies for novel anti-melanoma therapies. Adhesion Molecules in Normal Skin and Melanoma. Biomolecules 2021, 11, Keywords: cadherins; GTEx consortium; Human Protein Atlas; integrins; melanocytes; single cell 1213. https://doi.org/10.3390/ RNA sequencing; selectins; tumour microenvironment biom11081213 Academic Editor: Sang-Han Lee 1.
    [Show full text]
  • Mapping Transmembrane Binding Partners for E-Cadherin Ectodomains
    Mapping transmembrane binding partners for E-cadherin ectodomains Omer Shafraza, Bin Xieb, Soichiro Yamadaa, and Sanjeevi Sivasankara,b,1 aDepartment of Biomedical Engineering, University of California, Davis, CA 95616; and bBiophysics Graduate Group, University of California, Davis, CA 95616 Edited by Barry Honig, Howard Hughes Medical Institute, Columbia University, New York, NY, and approved October 28, 2020 (received for review May 22, 2020) We combine proximity labeling and single molecule binding assays proteins that directly interact with a transmembrane bait would to discover transmembrane protein interactions in cells. We first be positioned in close proximity to both the bait’s ectodomain screen for candidate binding partners by tagging the extracellular and cytoplasmic regions. Consequently, if the proximity-based and cytoplasmic regions of a “bait” protein with BioID biotin ligase labeling enzyme was fused to both the bait’s extracellular and and identify proximal proteins that are biotin tagged on both their cytoplasmic regions, identifying transmembrane proteins that extracellular and intracellular regions. We then test direct binding are biotinylated in both regions would enable us to narrow interactions between proximal proteins and the bait, using single down the list of possible binding partners for subsequent AFM molecule atomic force microscope binding assays. Using this ap- binding measurements. Furthermore, an integrated extracellu- proach, we identify binding partners for the extracellular region lar and cytoplasmic BioID measurement would significantly of E-cadherin, an essential cell–cell adhesion protein. We show that increase the precision of the screen by dramatically reducing the desmosomal proteins desmoglein-2 and desmocollin-3, the focal the number of false positive hits.
    [Show full text]
  • Pulmonary Neuroendocrine
    ORIGINAL RESEARCH published: 30 August 2021 doi: 10.3389/fonc.2021.645623 Pulmonary Neuroendocrine Neoplasms Overexpressing Epithelial-Mesenchymal Transition Mechanical Barriers Genes Lack Immune-Suppressive Response and Present an Edited by: Paul Takam Kamga, Increased Risk of Metastasis Universite´ de Versailles Saint-Quentin-en-Yvelines, France Tabatha Gutierrez Prieto 1*, Camila Machado Baldavira 1, Juliana Machado-Rugolo 1,2, Reviewed by: Cec´ılia Farhat 1, Eloisa Helena Ribeiro Olivieri 3, Vanessa Karen de Sa´ 3, ´ Tamas Zombori, Eduardo Caetano Abilio da Silva 4, Marcelo Luiz Balancin 1, Alexandre Muxfeldt Ab´Saber 1, University of Szeged, Hungary Teresa Yae Takagaki 5, Vladmir Cla´ udio Cordeiro de Lima 6,7 and Vera Luiza Capelozzi 1* Ryota Kurimoto, Tokyo Medical and Dental University, 1 Department of Pathology, University of São Paulo Medical School (USP), São Paulo, Brazil, 2 Health Technology Japan Assessment Center (NATS), Clinical Hospital (HCFMB), Medical School of São Paulo State University (UNESP), Helmut H. Popper, Botucatu, Brazil, 3 International Center of Research/CIPE, AC Camargo Cancer Center, São Paulo, Brazil, Medical University of Graz, Austria 4 Molecular Oncology Research Center, Barretos Cancer Hospital, Barretos, São Paulo, Brazil, 5 Division of Pneumology, *Correspondence: Instituto do Corac¸ão (Incor), Medical School of University of São Paulo, São Paulo, Brazil, 6 Oncology, Rede D’Or São Paulo, Tabatha Gutierrez Prieto São Paulo, Brazil, 7 Department of Clinical Oncology, Instituto do Caˆ ncer do Estado
    [Show full text]