Active Rab11 and functional recycling endosome are required for E-cadherin trafficking and lumen formation during epithelial morphogenesis

• Published in: American Journal of Physiology: Cell Physiology 2008-08, Vol.295 (2), p.C545-C556
• Main Authors: Desclozeaux, Marion, Venturato, Juliana, Wylie, Fiona G, Kay, Jason G, Joseph, Shannon R, Le, Huong T, Stow, Jennifer L
• Format: Article
• Language: English
• Subjects: Animals; Biochemistry; Cadherins - genetics; Cadherins - metabolism; Cell growth; Cell Line; Cell Polarity - physiology; Cells; Cysts; Cysts - metabolism; Cysts - pathology; Dogs; Endosomes - metabolism; Endosomes - physiology; Epithelial Cells - cytology; Epithelial Cells - metabolism; Epithelium - growth & development; Epithelium - metabolism; Green Fluorescent Proteins - genetics; Green Fluorescent Proteins - metabolism; Humans; Microscopy, Confocal; Morphogenesis - physiology; Mutation; Protein Transport - physiology; Proteins; rab GTP-Binding Proteins - genetics; rab GTP-Binding Proteins - metabolism; Receptors, Transferrin - genetics; Receptors, Transferrin - physiology; Recombinant Fusion Proteins - genetics; Recombinant Fusion Proteins - metabolism; Transfection; Transferrin - metabolism
• ISSN: 0363-6143; 1522-1563
• DOI: 10.1152/ajpcell.00097.2008

Institute for Molecular Bioscience, University of Queensland, St. Lucia, Queensland, Australia Submitted 17 February 2008 ; accepted in final form 21 June 2008 The correct targeting and trafficking of the adherens junction protein epithelial cadherin (E-cadherin) is a major determinant for the acquisition of epithelial cell polarity and for the maintenance of epithelial integrity. The compartments and trafficking components required to sort and transport E-cadherin to the basolateral cell surface remain to be fully defined. On the basis of previous data, we know that E-cadherin is trafficked via the recycling endosome (RE) in nonpolarized and newly polarized cells. Here we explore the role of the RE throughout epithelial morphogenesis in MDCK monolayers and cysts. Time-lapse microscopy in live cells, altering RE function biochemically, and expressing a dominant-negative form of Rab11 (DN-Rab11), each showed that the RE is always requisite for E-cadherin sorting and trafficking. The RE remained important for E-cadherin trafficking in MDCK cells from a nonpolarized state through to fully formed, polarized epithelial monolayers. During the development of epithelial cysts, DN-Rab11 disrupted E-cadherin targeting and trafficking, the subapical localization of pERM and actin, and cyst lumen formation. This final effect demonstrated an early and critical interdependence of Rab11 and the RE for E-cadherin targeting, apical membrane formation, and cell polarity in cysts. cyst; epithelia; cadherin; MDCK Address for reprint requests and other correspondence: J. L. Stow, Institute for Molecular Bioscience, University of Queensland, Brisbane 4072 Queensland, Australia (e-mail: j.stow{at}imb.uq.edu.au )