caa a22 4500 606176209 CHVBK 20210128100746.0 cr unu---uuuuu 210128e20151201xx s 000 0 eng 10.1007/s00018-015-1977-0 doi (NATIONALLICENCE)springer-10.1007/s00018-015-1977-0 E-cadherin and Src associate with extradesmosomal Dsg3 and modulate desmosome assembly and adhesion [Elektronische Daten] [Vera Rötzer, Eva Hartlieb, Franziska Vielmuth, Martin Gliem, Volker Spindler, Jens Waschke] Desmosomes provide strong intercellular cohesion essential for the integrity of cells and tissues exposed to continuous mechanical stress. For desmosome assembly, constitutively synthesized desmosomal cadherins translocate to the cell-cell border, cluster and mature in the presence of Ca2+ to stable cell contacts. As adherens junctions precede the formation of desmosomes, we investigated in this study the relationship between the classical cadherin E-cadherin and the desmosomal cadherin Desmoglein 3 (Dsg3), the latter of which is indispensable for cell-cell adhesion in keratinocytes. By using autoantibodies from patients with the blistering skin disease pemphigus vulgaris (PV), we showed in loss of function studies that E-cadherin compensates for effects of desmosomal disassembly. Overexpression of E-cadherin reduced the loss of cell cohesion induced by PV autoantibodies and attenuated activation of p38 MAPK. Silencing of E-cadherin abolished the localization of Dsg3 at the membrane and resulted in a shift of Dsg3 from the cytoskeletal to the non-cytoskeletal protein pool which conforms to the notion that E-cadherin regulates desmosome assembly. Mechanistically, we identified a complex consisting of extradesmosomal Dsg3, E-cadherin, β-catenin and Src and that the stability of this complex is regulated by Src. Moreover, Dsg3 and E-cadherin are phosphorylated on tyrosine residues in a Src-dependent manner and Src activity is required for recruiting Dsg3 to the cytoskeletal pool as well as for desmosome maturation towards a Ca2+-insensitive state. Our data provide new insights into the role of E-cadherin and the contribution of Src signaling for formation and maintenance of desmosomal junctions. Springer Basel, 2015 Desmosome nationallicence Src signaling nationallicence Desmoglein 3 (Dsg3) nationallicence E-cadherin nationallicence Calcium switch assay nationallicence Pemphigus nationallicence Dsg : Desmogleins nationallicence Dsc : Desmocollin nationallicence PV : Pemphigus vulgaris nationallicence PG : Plakoglobin nationallicence DP : Desmoplakin nationallicence E-cad : E-cadherin nationallicence Ab : Antibody nationallicence GFP : Green fluorescent protein nationallicence PKC : Protein kinase C nationallicence p38 MAPK : p38 mitogen-activated protein kinase nationallicence p-Tyr : Phospho-tyrosine nationallicence Src : Rous sarcoma (Src) kinase nationallicence Pkp : Plakophilin nationallicence AJ : Adherens junction nationallicence IgG : Immunoglobulin G nationallicence EGF : Epidermal growth factor nationallicence EGFR : Epidermal growth factor receptor nationallicence PBS : Phosphate buffered saline nationallicence Hbss : Hank's buffered saline solution nationallicence IP : Immunoprecipitation nationallicence siRNA : Small interfering RNA nationallicence n.t. : Non-targeting nationallicence Rötzer Vera Institute of Anatomy and Cell Biology, Department I, Ludwig-Maximilians-Universität, Munich, Germany aut Hartlieb Eva Institute of Anatomy and Cell Biology, Department I, Ludwig-Maximilians-Universität, Munich, Germany aut Vielmuth Franziska Institute of Anatomy and Cell Biology, Department I, Ludwig-Maximilians-Universität, Munich, Germany aut Gliem Martin Department of Ophthalmology, University of Bonn, Bonn, Germany aut Spindler Volker Institute of Anatomy and Cell Biology, Department I, Ludwig-Maximilians-Universität, Munich, Germany aut Waschke Jens Institute of Anatomy and Cell Biology, Department I, Ludwig-Maximilians-Universität, Munich, Germany aut Cellular and Molecular Life Sciences Springer International Publishing 72/24(2015-12-01), 4885-4897 1420-682X 72:24<4885 2015 72 18 https://doi.org/10.1007/s00018-015-1977-0 text/html Onlinezugriff via DOI BK010053 XK010053 XK010000 Metadata rights reserved Springer special CC-BY-NC licence nationallicence 1 research-article jats NATIONALLICENCE NATIONALLICENCE NL-springer NATIONALLICENCE 856 40 https://doi.org/10.1007/s00018-015-1977-0 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Rötzer Vera Institute of Anatomy and Cell Biology, Department I, Ludwig-Maximilians-Universität, Munich, Germany aut NATIONALLICENCE 700 1- Hartlieb Eva Institute of Anatomy and Cell Biology, Department I, Ludwig-Maximilians-Universität, Munich, Germany aut NATIONALLICENCE 700 1- Vielmuth Franziska Institute of Anatomy and Cell Biology, Department I, Ludwig-Maximilians-Universität, Munich, Germany aut NATIONALLICENCE 700 1- Gliem Martin Department of Ophthalmology, University of Bonn, Bonn, Germany aut NATIONALLICENCE 700 1- Spindler Volker Institute of Anatomy and Cell Biology, Department I, Ludwig-Maximilians-Universität, Munich, Germany aut NATIONALLICENCE 700 1- Waschke Jens Institute of Anatomy and Cell Biology, Department I, Ludwig-Maximilians-Universität, Munich, Germany aut NATIONALLICENCE 773 0- Cellular and Molecular Life Sciences Springer International Publishing 72/24(2015-12-01), 4885-4897 1420-682X 72:24<4885 2015 72 18