Article
- The EMBO Journal (2007) 26, 3737 - 3748
- doi:10.1038/sj.emboj.7601813
Published online: 2 August 2007
Subject Category:
Differential involvement of endocytic compartments in the biosynthetic traffic of apical proteins
Kerry O Cresawn1,a, Beth A Potter1,a, Asli Oztan1, Christopher J Guerriero1, Gudrun Ihrke3, James R Goldenring4, Gerard Apodaca1,2 and Ora A Weisz1,2
- Renal-Electrolyte Division, University of Pittsburgh, Pittsburgh, PA, USA
- Department of Cell Biology and Physiology, University of Pittsburgh, Pittsburgh, PA, USA
- Department of Pharmacology, Uniformed Services University of the Health Sciences, Bethesda, MD, USA
- Department of Surgery, Vanderbilt University School of Medicine and Nashville Veterans Affairs Medical Center, Nashville, TN, USA
Correspondence to:
Ora A Weisz, Renal-Electrolyte Division, University of Pittsburgh, 3550 Terrace St., Pittsburgh, PA 15261, USA. Tel.: +1 412 383 8891; Fax: +1 412 383 8956; E-mail: weisz@pitt.edu
aThese authors equally contributed to this work
Received 12 January 2007; Accepted 4 July 2007
Abstract
Newly synthesized basolateral markers can traverse recycling endosomes en route to the surface of Madin–Darby canine kidney cells; however, the routes used by apical proteins are less clear. Here, we functionally inactivated subsets of endocytic compartments and examined the effect on surface delivery of the basolateral marker vesicular stomatitis virus glycoprotein (VSV-G), the raft-associated apical marker influenza hemagglutinin (HA), and the non-raft-associated protein endolyn. Inactivation of transferrin-positive endosomes after internalization of horseradish peroxidase (HRP)-containing conjugates inhibited VSV-G delivery, but did not disrupt apical delivery. In contrast, inhibition of protein export from apical recycling endosomes upon expression of dominant-negative constructs of myosin Vb or Sec15 selectively perturbed apical delivery of endolyn. Ablation of apical endocytic components accessible to HRP-conjugated wheat germ agglutinin (WGA) disrupted delivery of HA but not endolyn. However, delivery of glycosylphosphatidylinositol-anchored endolyn was inhibited by >50% under these conditions, suggesting that the biosynthetic itinerary of a protein is dependent on its targeting mechanism. Our studies demonstrate that apical and basolateral proteins traverse distinct endocytic intermediates en route to the cell surface, and that multiple routes exist for delivery of newly synthesized apical proteins.
Keywords:
- biosynthetic delivery,
- endosome,
- lipid raft,
- MDCK,
- polarized
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