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© The Rockefeller University Press, 0021-9525/1998//611 $5.00
The Journal of Cell Biology, Volume 141, Number 3, , 1998 611-623

Articles

In Polarized MDCK Cells Basolateral Vesicles Arise from Clathrin-{gamma}-adaptin–coated Domains on Endosomal Tubules



C.E. Futter*, A. Gibson*, E.H. Allchin*, S. Maxwell*, L.J. Ruddock*, G. Odorizzi{ddagger}, D. Domingo{ddagger}, I.S. Trowbridge{ddagger}, and C.R. Hopkins*

* Medical Research Council Laboratory for Molecular Cell Biology, University College London, London WC1E 6BT, England; and {ddagger} Department of Cancer Biology, The Salk Institute for Biological Studies, San Diego, California 92186-5800

Human transferrin receptors (TR) and receptors for polymeric immunoglobulins (pIgR) expressed in polarized MDCK cells maintain steady-state, asymmetric distributions on the separate basolateral and apical surfaces even though they are trafficking continuously into and across these cells. The intracellular mechanisms required to maintain these asymmetric distributions have not been located. Here we show that TR and pIgR internalize from both surfaces to a common interconnected endosome compartment that includes tubules with buds coated with clathrin lattices. These buds generate vesicles that carry TR to the basolateral border. The lattices contain {gamma}-adaptin and are dispersed by treatment with brefeldin A (BFA). Since BFA treatment abrogates the vectorial trafficking of TR in polarized MDCK cells, we propose that the clathrin-coated domains of the endosome tubules contain the polarized sorting mechanism responsible for their preferential basolateral distribution.

Abbreviations used in this paper: IgA, dimeric IgA; BFA, brefeldin A; LDL-R, low-density lipoprotein receptor; PB, permeabilization buffer; pIgR, polymeric immunoglobulin receptor; ST-HRP, sialyl transferase-HRP; TF, transferrin; TR, transferrin receptor(s).

Address all correspondence to Colin R. Hopkins, Medical Research Council Laboratory for Molecular Cell Biology, University College London, Gordon St., London WC1E 6BT, England. Tel.: 0171-380-7806. Fax: 0171-380-7804.


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