Trafficking of Crumbs3 during cytokinesis is crucial for lumen formation

Marc A Schlüter, Catherine S Pfarr, Jay Pieczynski, Eileen L Whiteman, Toby W Hurd, Shuling Fan, Chia-Jen Liu, Ben Margolis

Research output: Contribution to journalArticlepeer-review


Although lumen generation has been extensively studied through so-called cyst-formation assays in Madin-Darby canine kidney (MDCK) cells, an underlying mechanism that leads to the initial appearance of a solitary lumen remains elusive. Lumen formation is thought to take place at early stages in aggregates containing only a few cells. Evolutionarily conserved polarity protein complexes, namely the Crumbs, Par, and Scribble complexes, establish apicobasal polarity in epithelial cells, and interference with their function impairs the regulated formation of solitary epithelial lumina. Here, we demonstrate that MDCK cells form solitary lumina during their first cell division. Before mitosis, Crumbs3a becomes internalized and concentrated in Rab11-positive recycling endosomes. These compartments become partitioned in both daughter cells and are delivered to the site of cytokinesis, thus forming the first apical membrane, which will eventually form a lumen. Endosome trafficking in this context appears to depend on the mitotic spindle apparatus and midzone microtubules. Furthermore, we show that this early lumen formation is regulated by the apical polarity complexes because Crumbs3 assists in the recruitment of aPKC to the forming apical membrane and interference with their function can lead to the formation of a no-lumen or multiple-lumen phenotype at the two-cell stage.
Original languageEnglish
Pages (from-to)4652-63
Number of pages12
JournalMolecular Biology of the Cell
Issue number22
Publication statusPublished - Nov 2009


  • Animals
  • Cell Cycle Proteins
  • Cell Line
  • Cell Polarity
  • Cytokinesis
  • Cytoskeleton
  • Dogs
  • Endocytosis
  • Endosomes
  • Membrane Proteins
  • Microtubules
  • Morphogenesis
  • Multiprotein Complexes
  • Protein Isoforms
  • Protein Kinase C
  • Protein Transport
  • Recombinant Fusion Proteins
  • rab GTP-Binding Proteins


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