Alpha-catenin-dependent recruitment of the centrosomal protein CAP350 to adherens junctions allows epithelial cells to acquire a columnar shape

Maria P Gavilan; Marina Arjona; Angel Zurbano; Etienne Formstecher; Juan R Martinez-Morales; Michel Bornens; Rosa M Rios

doi:10.1371/journal.pbio.1002087

Alpha-catenin-dependent recruitment of the centrosomal protein CAP350 to adherens junctions allows epithelial cells to acquire a columnar shape

PLoS Biol. 2015 Mar 12;13(3):e1002087. doi: 10.1371/journal.pbio.1002087. eCollection 2015 Mar.

Authors

Maria P Gavilan¹, Marina Arjona¹, Angel Zurbano¹, Etienne Formstecher², Juan R Martinez-Morales³, Michel Bornens⁴, Rosa M Rios¹

Affiliations

¹ Centro Andaluz de Biología Molecular y Medicina Regenerativa (CABIMER), CSIC, Sevilla, Spain.
² Hybrigenics Services SA, Paris, France.
³ Centro Andaluz de Biología del Desarrollo, CSIC/UPO/JA, Sevilla, Spain.
⁴ UMR144 CNRS-Institut Curie, Paris, France.

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.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Adherens Junctions / metabolism
Adherens Junctions / ultrastructure
Adipocytes / cytology
Adipocytes / metabolism
Animals
Cadherins / genetics
Cadherins / metabolism
Cell Adhesion
Cell Line
Cell Polarity
Cell Shape
Dogs
Embryo, Nonmammalian
Epithelial Cells / cytology
Epithelial Cells / metabolism*
Gene Expression Regulation, Developmental*
Genetic Vectors
Humans
Lentivirus / genetics
Madin Darby Canine Kidney Cells
Microtubule Proteins / antagonists & inhibitors
Microtubule Proteins / genetics*
Microtubule Proteins / metabolism
Microtubules / metabolism
Microtubules / ultrastructure
Morphogenesis / genetics*
Nuclear Proteins / antagonists & inhibitors
Nuclear Proteins / genetics*
Nuclear Proteins / metabolism
Oryzias
RNA, Small Interfering / genetics
RNA, Small Interfering / metabolism
Signal Transduction
Two-Hybrid System Techniques
alpha Catenin / genetics*
alpha Catenin / metabolism

Substances

CEP350 protein, human
Cadherins
Microtubule Proteins
Nuclear Proteins
RNA, Small Interfering
alpha Catenin

Grants and funding

This work was supported by Ministerio de Economia y Competitividad, Spain (BFU2012-36717 and CSD2009-00016 to RMR and BFU2011-22916 to JRM) and by Junta de Andalucia (CVI-7256 and CTS-2071), and by a funding GenHomme Network 02490-6088 to Hybrigenics and the Institut Curie. MA and AZ were supported by MEC–FPI Grants. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.