title: Drebrin E depletion in human intestinal epithelial cells mimics Rab8a loss of function.
abstract: Intestinal epithelial cells are highly polarized and exhibit a complex architecture with a columnar shape and a specialized apical surface supporting microvilli organized in a brush border. These microvilli are rooted in a dense meshwork of acto-myosin called the terminal web. We have shown recently that Drebrin E, an F-actin-binding protein, is a key protein for the organization of the terminal web and the brush border. Drebrin E is also required for the columnar cell shape of Caco2 cells (human colonic cells). Here, we found that the subcellular localization of several apical markers including dipeptidyl peptidase IV (DPPIV) was strikingly modified in Drebrin E-depleted Caco2 cells. Instead of being mostly present at the apical surface, these proteins are accumulated in an enlarged subapical compartment. Using known intracellular markers, we show by both confocal and electron microscopy that this compartment is related to lysosomes. We also demonstrate that the enrichment of DPPIV in this compartment originates from apical endocytosis and that depletion of Rab8a induces an accumulation of apical proteins in a similar compartment. Consistent with this, the phenotype observed in Drebrin E knock-down Caco2 cells shares some features with a pathology called microvillar inclusion disease (MVID) involving both Myosin Vb and Rab8a. Taken together, these results suggest that Drebrin E redirects the apical recycling pathway in intestinal epithelial cells to the lysosomes, demonstrating that Drebrin E is a key regulator in apical trafficking in Caco2 cells.
authors.affiliation: Aix-Marseille University, CNRS, UMR7288, Developmental Biology Institute of Marseille (IBDM), Case 907, 13288 Marseille, Cedex 09, France.
authors.forename: Barbara
authors.initials: B
authors.lastname: Vacca
authors.forename: Elsa
authors.initials: E
authors.lastname: Bazellières
authors.forename: Roqiya
authors.initials: R
authors.lastname: Nouar
authors.forename: Akihiro
authors.initials: A
authors.lastname: Harada
authors.forename: Dominique
authors.initials: D
authors.lastname: Massey-Harroche
authors.forename: André
authors.initials: A
authors.lastname: Le Bivic
mesh_terms: D018938:Caco-2 Cells; D016764:Cell Polarity; D004705:Endocytosis; D004847:Epithelial Cells / Q000378:metabolism*; D055785:Gene Knockdown Techniques; D006801:Humans; D007413:Intestinal Mucosa / Q000378:metabolism*; D007422:Intestines / Q000166:cytology; D008871:Microvilli / Q000235:genetics / Q000378:metabolism; D009479:Neuropeptides / Q000172:deficiency* / Q000235:genetics; D021381:Protein Transport; D020691:rab GTP-Binding Proteins / Q000235:genetics / Q000378:metabolism*
chemical_list: D009479:Neuropeptides; C053386:drebrins; C082377:RAB8A protein, human; D020691:rab GTP-Binding Proteins