Iterative sorting of apical and basolateral cargo in Madin-Darby canine kidney cells.

Abstract:

:For several decades, the trans-Golgi network (TGN) was considered the most distal stop and hence the ultimate protein-sorting station for distinct apical and basolateral transport carriers that reach their respective surface domains in the direct trafficking pathway. However, recent reports of apical and basolateral cargoes traversing post-Golgi compartments accessible to endocytic ligands before their arrival at the cell surface and the post-TGN breakup of large pleomorphic membrane fragments that exit the Golgi region toward the surface raised the possibility that compartments distal to the TGN mediate or contribute to biosynthetic sorting. Here we describe the development of a novel assay that quantitatively distinguishes different cargo pairs by their degree of colocalization at the TGN and by the evolution of colocalization during their TGN-to-surface transport. Keys to the high resolution of our approach are 1) conversion of perinuclear organelle clustering into a two-dimensional microsomal spread and 2) identification of TGN and post-TGN cargo without the need for a TGN marker that universally cosegregates with all cargo. Using our assay, we provide the first evidence that apical NTRp75 and basolateral VSVG in Madin-Darby canine kidney cells still undergo progressive sorting after they exit the TGN toward the cell surface.

journal_name

Mol Biol Cell

authors

Treyer A,Pujato M,Pechuan X,Müsch A

doi

10.1091/mbc.E16-02-0096

subject

Has Abstract

pub_date

2016-07-15 00:00:00

pages

2259-71

issue

14

eissn

1059-1524

issn

1939-4586

pii

mbc.E16-02-0096

journal_volume

27

pub_type

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