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Published online
doi:10.1083/jcb.200901021
The Journal of Cell Biology, Vol. 186, No. 2, 269-282
The Rockefeller University Press, 0021-9525 $30.00
© Farr et al.
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Article

Membrane proteins follow multiple pathways to the basolateral cell surface in polarized epithelial cells



Glen A. Farr1,2, Michael Hull1,2, Ira Mellman3, and Michael J. Caplan1,2

1 Department of Cellular and Molecular Physiology and 2 Department of Cell Biology, Yale University School of Medicine, New Haven, CT 06520
3 Genentech, Inc., South San Francisco, CA 94080

Correspondence to Michael J. Caplan: michael.caplan{at}yale.edu

Newly synthesized apical and basolateral membrane proteins are sorted from one another in polarized epithelial cells. The trans-Golgi network participates in this sorting process, but some basolateral proteins travel from the Golgi to recycling endosomes (REs) before their surface delivery. Using a novel system for pulse–chase microscopy, we have visualized the postsynthetic route pursued by a newly synthesized cohort of Na,K-ATPase. We find that the basolateral delivery of newly synthesized Na,K-ATPase occurs via a pathway distinct from that pursued by the vesicular stomatitis virus G protein (VSV-G). Na,K-ATPase surface delivery occurs at a faster rate than that observed for VSV-G. The Na,K-ATPase does not pass through the RE compartment en route to the plasma membrane, and Na,K-ATPase trafficking is not regulated by the same small GTPases as other basolateral proteins. Finally, Na,K-ATPase and VSV-G travel in separate post-Golgi transport intermediates, demonstrating directly that multiple routes exist for transport from the Golgi to the basolateral membrane in polarized epithelial cells.


Abbreviations used in this paper: BG, benzylguanine; CHX, cycloheximide; E-cadherin, epithelial cadherin; LDLR, low density lipoprotein receptor; PGTI, post-Golgi transport intermediate; PM, plasma membrane; RE, recycling endosome; shRNA, short hairpin RNA; Tfn, transferrin; Tfn-R, Tfn receptor; VSV-G, vesicular stomatitis virus G protein.

© 2009 Farr et al.
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