title: The N-glycan processing in HT-29 cells is a function of their state of enterocytic differentiation. Evidence for an atypical traffic associated with change in polypeptide stability in undifferentiated HT-29 cells.
abstract: When the human colon cancer cells HT-29 undergo enterocytic differentiation, they correctly process their N-glycans, whereas their undifferentiated counterpart are unable to process Man9-8-GlcNAc2 species, the natural substrate of alpha-mannosidase I. As this enzyme is fully active in both HT-29 cell populations, we hypothesize that N-glycoproteins are unable to reach the cis Golgi, the site where alpha-mannosidase I has been localized. We have demonstrated this point by using 1-deoxymannojirimycin, leupeptin, and monensin. In the presence of 1-deoxymannojirimycin, a specific inhibitor of alpha-mannosidase I, differentiated HT-29 cells, as expected, accumulate Man9-8-GlcNAc2 species, whereas in undifferentiated HT-29 cells these compounds continue to be rapidly degraded. In contrast, the use of leupeptin, a specific inhibitor of thiol and serine proteases, leads to the accumulation of these oligosaccharides in undifferentiated HT-29 cells. Monensin, a carboxylic ionophore that perturbs distal Golgi functions, is unable to stabilize these compounds. Therefore, we conclude that N-linked glycoproteins in undifferentiated HT-29 cells rapidly egress from the exocytic pathway to a leupeptin-sensitive degradative compartment without entering a monensin-sensitive compartment. These results favor the hypothesis that a direct pathway should exist between the rough endoplasmic reticulum and a leupeptin-sensitive degradative compartment in undifferentiated HT-29 cells. The emergence of this new pathway could explain why protein stability and N-glycan processing may vary as a function of the state of cell differentiation.
authors.affiliation: Unité de Recherche sur la Différenciation et la Neuroendocrinologie des Cellules Digestives, Institut National de la Santé et de la Recherche Médicale U 178, Villejuif, France.
authors.forename: G
authors.initials: G
authors.lastname: Trugnan
authors.forename: E
authors.initials: E
authors.lastname: Ogier-Denis
authors.forename: C
authors.initials: C
authors.lastname: Sapin
authors.forename: D
authors.initials: D
authors.lastname: Darmoul
authors.forename: C
authors.initials: C
authors.lastname: Bauvy
authors.forename: M
authors.initials: M
authors.lastname: Aubery
authors.forename: P
authors.initials: P
authors.lastname: Codogno
mesh_terms: D017485:1-Deoxynojirimycin; D001692:Biological Transport; D002454:Cell Differentiation; D003106:Colon / Q000166:cytology / Q000378:metabolism*; D003110:Colonic Neoplasms; D005944:Glucosamine / Q000031:analogs & derivatives / Q000494:pharmacology; D006801:Humans; D066298:In Vitro Techniques; D007976:Leupeptins / Q000494:pharmacology; D008361:Mannosidases / Q000378:metabolism*; D008562:Membrane Glycoproteins / Q000378:metabolism*; D008985:Monensin / Q000494:pharmacology; D011134:Polysaccharides / Q000378:metabolism*; D011499:Protein Processing, Post-Translational / Q000187:drug effects; D014407:Tumor Cells, Cultured; D043323:alpha-Mannosidase
chemical_list: D007976:Leupeptins; D008562:Membrane Glycoproteins; D011134:Polysaccharides; C025363:mannoproteins; D017485:1-Deoxynojirimycin; D008985:Monensin; D008361:Mannosidases; D043323:alpha-Mannosidase; C032854:leupeptin; D005944:Glucosamine