Microtubule perturbation retards both the direct and the indirect apical pathway but does not affect sorting of plasma membrane proteins in intestinal epithelial cells (Caco-2).
Endogenous plasma membrane proteins are sorted from two sites in the human intestinal epithelial cell line Caco-2. Apical proteins are transported from the Golgi apparatus to the apical domain along a direct pathway and an indirect pathway via the basolateral membrane. In contrast, basolateral proteins never appear in the apical plasma membrane. Here we report on the effect of the microtubule-active drug nocodazole on the post-synthetic transport and sorting of plasma membrane proteins. Pulse-chase radiolabeling was combined with domain-specific cell surface assays to monitor the appearance of three apical and one basolateral protein in plasma membrane domains. Nocodazole was found to drastically retard both the direct transport of apical proteins from the Golgi apparatus and the indirect transport (transcytosis) from the basolateral membrane to the apical cell surface. In contrast, neither the transport rates of the basolateral membrane nor the sorting itself were significantly affected by the nocodazole treatment. We conclude that an intact microtubular network facilitates, but is not necessarily required for, the transport of apical membrane proteins along the two post-Golgi pathways to the brush border.
Department of Pharmacology, Biocenter of the University of Basel, Switzerland.
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D000230:Adenocarcinoma; D002460:Cell Line; D002462:Cell Membrane / Q000502:physiology*; D003110:Colonic Neoplasms; D004705:Endocytosis / Q000187:drug effects; D004848:Epithelium; D006801:Humans; D007700:Kinetics; D008562:Membrane Glycoproteins / Q000096:biosynthesis*; D008565:Membrane Proteins / Q000096:biosynthesis*; D008715:Methionine / Q000378:metabolism; D008870:Microtubules / Q000502:physiology* / Q000648:ultrastructure; D015739:Nocodazole / Q000494:pharmacology; D014404:Tubulin / Q000378:metabolism
D008562:Membrane Glycoproteins; D008565:Membrane Proteins; D014404:Tubulin; D008715:Methionine; D015739:Nocodazole