The Journal of Cell Biology, Vol 108, 13-22, Copyright © 1989 by The Rockefeller University Press

ARTICLES

Nocodazole, a microtubule-active drug, interferes with apical protein delivery in cultured intestinal epithelial cells (Caco-2)

U Eilers, J Klumperman and HP Hauri
Department of Pharmacology, Biocenter of the University of Basel, Switzerland.

The polarized delivery of membrane proteins to the cell surface and the initial secretion of lysosomal proteins into the culture medium were studied in the polarized human intestinal adenocarcinoma cell line Caco- 2 in the presence or absence of the microtubule-active drug nocodazole. The appearance of newly synthesized proteins at the plasma membrane was measured by their sensitivity to proteases added either to the apical or the basolateral surface of cells grown on nitrocellulose filters. Nocodazole was found to reduce the delivery to the cell surface of an apical membrane protein, aminopeptidase N, and to lead to its partial missorting to the basolateral surface, whereas the drug had no influence on the delivery of a basolateral 120-kD membrane protein defined by a monoclonal antibody. Furthermore, nocodazole selectively blocked the apical secretion of two lysosomal proteins, cathepsin D and acid alpha-glucosidase, whereas the drug had no influence on their basolateral secretion. These results suggest that in Caco-2 cells an intact microtubular network is important for the transport of newly synthesized proteins to the apical cell surface.
CiteULike    Complore    Connotea    Del.icio.us    Digg    Facebook    Reddit    Technorati    Twitter    What's this?

This article has been cited by other articles:

• Driss, A., Charrier, L., Yan, Y., Nduati, V., Sitaraman, S., Merlin, D. (2006). Dystroglycan receptor is involved in integrin activation in intestinal epithelia. Am. J. Physiol. Gastrointest. Liver Physiol. 290: G1228-G1242 [Abstract] [Full Text]  
• Lim, C. M., Cater, M. A., Mercer, J. F. B., La Fontaine, S. (2006). Copper-dependent Interaction of Dynactin Subunit p62 with the N Terminus of ATP7B but Not ATP7A. J. Biol. Chem. 281: 14006-14014 [Abstract] [Full Text]  
• Marozin, S., Prank, U., Sodeik, B. (2004). Herpes simplex virus type 1 infection of polarized epithelial cells requires microtubules and access to receptors present at cell-cell contact sites. J. Gen. Virol. 85: 775-786 [Abstract] [Full Text]  
• TUMA, P. L., HUBBARD, A. L. (2003). Transcytosis: Crossing Cellular Barriers. Physiol. Rev. 83: 871-932 [Abstract] [Full Text]  
• Chu, J. J. H., Ng, M. L. (2002). Infection of polarized epithelial cells with flavivirus West Nile: polarized entry and egress of virus occur through the apical surface. J. Gen. Virol. 83: 2427-2435 [Abstract] [Full Text]  
• Wick, D. A., Seetharam, B., Dahms, N. M. (1999). Biosynthesis and secretion of the mannose 6-phosphate receptor and its ligands in polarized Caco-2 cells. Am. J. Physiol. Gastrointest. Liver Physiol. 277: G506-G514 [Abstract] [Full Text]  
• YEAMAN, C., GRINDSTAFF, K. K., NELSON, W. J. (1999). New Perspectives on Mechanisms Involved in Generating Epithelial Cell Polarity. Physiol. Rev. 79: 73-98 [Abstract] [Full Text]  
• Zegers, M. M. P., Zaal, K. J. M., van IJzendoorn, S. C. D., Klappe, K., Hoekstra, D. (1998). Actin Filaments and Microtubules are Involved in Different Membrane Traffic Pathways That Transport Sphingolipids to the Apical Surface of Polarized HepG2 Cells. Mol. Biol. Cell 9: 1939-1949 [Abstract] [Full Text]  
• Huet, G., Hennebicq-Reig, S., de Bolos, C., Ulloa, F., Lesuffleur, T., Barbat, A., Carriere, V., Kim, I., Real, F. X., Delannoy, P., Zweibaum, A. (1998). GalNAc-{alpha}-O-benzyl Inhibits NeuAc{alpha}2-3 Glycosylation and Blocks the Intracellular Transport of Apical Glycoproteins and Mucus in Differentiated HT-29 Cells. JCB 141: 1311-1322 [Abstract] [Full Text]  
• Grindstaff, K. K., Bacallao, R. L., Nelson, W. J. (1998). Apiconuclear Organization of Microtubules Does Not Specify Protein Delivery from the Trans-Golgi Network to Different Membrane Domains in Polarized Epithelial Cells. Mol. Biol. Cell 9: 685-699 [Abstract] [Full Text]  
• Saunders, C., Limbird, L. E. (1997). Disruption of Microtubules Reveals Two Independent Apical Targeting Mechanisms for G-protein-coupled Receptors in Polarized Renal Epithelial Cells. J. Biol. Chem. 272: 19035-19045 [Abstract] [Full Text]  
• Abu-Amer, Y., Ross, F. P., Schlesinger, P., Tondravi, M. M., Teitelbaum, S. L. (1997). Substrate Recognition by Osteoclast Precursors Induces C-src/Microtubule Association. JCB 137: 247-258 [Abstract] [Full Text]  
• Bulinski, J., McGraw, T., Gruber, D, Nguyen, H., Sheetz, M. (1997). Overexpression of MAP4 inhibits organelle motility and trafficking in vivo. J. Cell Sci. 110: 3055-3064 [Abstract]  
• Mayer, A, Ivanov, I., Gravotta, D, Adesnik, M, Sabatini, D. (1996). Cell-free reconstitution of the transport of viral glycoproteins from the TGN to the basolateral plasma membrane of MDCK cells. J. Cell Sci. 109: 1667-1676 [Abstract]  
• Tousson, A, Fuller, C., Benos, D. (1996). Apical recruitment of CFTR in T-84 cells is dependent on cAMP and microtubules but not Ca2+ or microfilaments. J. Cell Sci. 109: 1325-1334 [Abstract]  
• Arreaza, G., Brown, D. A. (1995). Sorting and Intracellular Trafficking of a Glycosylphosphatidylinositol-anchored Protein and Two Hybrid Transmembrane Proteins with the Same Ectodomain in Madin-Darby Canine Kidney Epithelial Cells. J. Biol. Chem. 270: 23641-23647 [Abstract] [Full Text]  
• Baricault, L, Fransen, J., Garcia, M, Sapin, C, Codogno, P, Ginsel, L., Trugnan, G (1995). Rapid sequestration of DPP IV/CD26 and other cell surface proteins in an autophagic-like compartment in Caco-2 cells treated with forskolin. J. Cell Sci. 108: 2109-2121 [Abstract]  
• Chou, C. F., Omary, M. B. (1994). Mitotic arrest with anti-microtubule agents or okadaic acid is associated with increased glycoprotein terminal GlcNAc's. J. Cell Sci. 107: 1833-1843 [Abstract]  
• Appel, D, Koch-Brandt, C (1994). Sorting of a secretory protein (gp80) to the apical surface of Caco-2 cells. J. Cell Sci. 107: 553-559 [Abstract]  
• Hamm-Alvarez, S., Kim, P., Sheetz, M. (1993). Regulation of vesicle transport in CV-1 cells and extracts. J. Cell Sci. 106: 955-966 [Abstract]  
• van Meer, G, van 't Hof, W (1993). Epithelial sphingolipid sorting is insensitive to reorganization of the Golgi by nocodazole, but is abolished by monensin in MDCK cells and by brefeldin A in Caco-2 cells. J. Cell Sci. 104: 833-842 [Abstract]  
• Burkhardt, J., McIlvain, J., Sheetz, M., Argon, Y (1993). Lytic granules from cytotoxic T cells exhibit kinesin-dependent motility on microtubules in vitro. J. Cell Sci. 104: 151-162 [Abstract]  
• Ojakian, G. K., Schwimmer, R. (1992). Antimicrotubule drugs inhibit the polarized insertion of an intracellular glycoprotein pool into the apical membrane of Madin-Darby canine kidney (MDCK) cells. J. Cell Sci. 103: 677-687 [Abstract]  
• Rodriguez-Boulan, E, Nelson, W. (1989). Morphogenesis of the polarized epithelial cell phenotype. Science 245: 718-725 [Abstract]  

Home | Help | Feedback | Subscriptions | Archive | Search | Table of Contents