title: Alpha-catenin-dependent recruitment of the centrosomal protein CAP350 to adherens junctions allows epithelial cells to acquire a columnar shape.
abstract: Epithelial morphogenesis involves a dramatic reorganisation of the microtubule cytoskeleton. How this complex process is controlled at the molecular level is still largely unknown. Here, we report that the centrosomal microtubule (MT)-binding protein CAP350 localises at adherens junctions in epithelial cells. By two-hybrid screening, we identified a direct interaction of CAP350 with the adhesion protein α-catenin that was further confirmed by co-immunoprecipitation experiments. Block of epithelial cadherin (E-cadherin)-mediated cell-cell adhesion or α-catenin depletion prevented CAP350 localisation at cell-cell junctions. Knocking down junction-located CAP350 inhibited the establishment of an apico-basal array of microtubules and impaired the acquisition of columnar shape in Madin-Darby canine kidney II (MDCKII) cells grown as polarised epithelia. Furthermore, MDCKII cystogenesis was also defective in junctional CAP350-depleted cells. CAP350-depleted MDCKII cysts were smaller and contained either multiple lumens or no lumen. Membrane polarity was not affected, but cortical microtubule bundles did not properly form. Our results indicate that CAP350 may act as an adaptor between adherens junctions and microtubules, thus regulating epithelial differentiation and contributing to the definition of cell architecture. We also uncover a central role of α-catenin in global cytoskeleton remodelling, in which it acts not only on actin but also on MT reorganisation during epithelial morphogenesis.
authors.affiliation: Centro Andaluz de Biología Molecular y Medicina Regenerativa (CABIMER), CSIC, Sevilla, Spain.
authors.forename: Maria P
authors.initials: MP
authors.lastname: Gavilan
authors.affiliation: Centro Andaluz de Biología Molecular y Medicina Regenerativa (CABIMER), CSIC, Sevilla, Spain.
authors.forename: Marina
authors.initials: M
authors.lastname: Arjona
authors.affiliation: Centro Andaluz de Biología Molecular y Medicina Regenerativa (CABIMER), CSIC, Sevilla, Spain.
authors.forename: Angel
authors.initials: A
authors.lastname: Zurbano
authors.affiliation: Hybrigenics Services SA, Paris, France.
authors.forename: Etienne
authors.initials: E
authors.lastname: Formstecher
authors.affiliation: Centro Andaluz de Biología del Desarrollo, CSIC/UPO/JA, Sevilla, Spain.
authors.forename: Juan R
authors.initials: JR
authors.lastname: Martinez-Morales
authors.affiliation: UMR144 CNRS-Institut Curie, Paris, France.
authors.forename: Michel
authors.initials: M
authors.lastname: Bornens
authors.affiliation: Centro Andaluz de Biología Molecular y Medicina Regenerativa (CABIMER), CSIC, Sevilla, Spain.
authors.forename: Rosa M
authors.initials: RM
authors.lastname: Rios
mesh_terms: D022005:Adherens Junctions / Q000378:metabolism / Q000648:ultrastructure; D017667:Adipocytes / Q000166:cytology / Q000378:metabolism; D000818:Animals; D015820:Cadherins / Q000235:genetics / Q000378:metabolism; D002448:Cell Adhesion; D002460:Cell Line; D016764:Cell Polarity; D048430:Cell Shape; D004285:Dogs; D004625:Embryo, Nonmammalian; D004847:Epithelial Cells / Q000166:cytology / Q000378:metabolism*; D018507:Gene Expression Regulation, Developmental; D005822:Genetic Vectors; D006801:Humans; D016086:Lentivirus / Q000235:genetics; D061985:Madin Darby Canine Kidney Cells; D008868:Microtubule Proteins / Q000037:antagonists & inhibitors / Q000235:genetics* / Q000378:metabolism; D008870:Microtubules / Q000378:metabolism / Q000648:ultrastructure; D009024:Morphogenesis / Q000235:genetics*; D009687:Nuclear Proteins / Q000037:antagonists & inhibitors / Q000235:genetics* / Q000378:metabolism; D009990:Oryzias; D034741:RNA, Small Interfering / Q000235:genetics / Q000378:metabolism; D015398:Signal Transduction; D020798:Two-Hybrid System Techniques; D051178:alpha Catenin / Q000235:genetics* / Q000378:metabolism
chemical_list: C509238:CEP350 protein, human; D015820:Cadherins; D008868:Microtubule Proteins; D009687:Nuclear Proteins; D034741:RNA, Small Interfering; D051178:alpha Catenin