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J Biol Chem, Vol. 275, Issue 20, 15207-15219, May 19, 2000

Interactions between the Exocytic and Endocytic Pathways in Polarized Madin-Darby Canine Kidney Cells*

Ena OrzechDagger , Shulamit CohenDagger , Aryeh Weiss§, and Benjamin AroetiDagger

From the Dagger  Department of Cell and Animal Biology, Institute of Life Sciences, Hebrew University of Jerusalem, Jerusalem 91904 and the § Department of Electronics, Jerusalem College of Technology, Jerusalem 91160, Israel

The compartments involved in polarized exocytosis of membrane proteins are not well defined. In this study we hypothesized that newly synthesized polymeric immunoglobulin receptors are targeted from the trans-Golgi network to endosomes prior to their appearance on the basolateral cell surface of polarized Madin-Darby canine kidney cells. To examine this hypothesis, we have used an assay designed to measure the meeting of newly synthesized receptors with a selective population of apical or basolateral endosomes loaded with horseradish peroxidase. We found that in the course of basolateral exocytosis, the wild-type polymeric immunoglobulin receptor is targeted from the trans-Golgi network to apical and basolateral endosomes. Phosphorylation of a Ser residue in the cytoplasmic tail of the receptor is implicated in this process. The biosynthetic pathway of apically sorted polymeric immunoglobulin receptor mutants similarly traversed apical endosomes, raising the possibility that apical receptors are segregated from basolateral receptors in apical endosomes. The post-endocytic pathway of transcytosing and recycling receptors also passed through apical endosomes. Together, these observations are consistent with the possibility that the biosynthetic and endocytic routes merge into endosomes and justify a model suggesting that endosomal recycling processes govern polarized trafficking of proteins traveling in both pathways.

* This work was supported by a grant from the Israel Science Foundation founded by the Israel Academy of Sciences.The costs of publication of this article were defrayed in part by the payment of page charges. The article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

To whom correspondence should be addressed. Fax: 972-2-5617918; E-mail: aroeti@cc.huji.ac.il.

Copyright © 2000 by The American Society for Biochemistry and Molecular Biology, Inc.
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