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Originally published as MBC in Press, 10.1091/mbc.E08-12-1229 on August 26, 2009

Vol. 20, Issue 20, 4458-4470, October 15, 2009

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The Function of the Intermediate Compartment in Pre-Golgi Trafficking Involves its Stable Connection with the Centrosome

Michaël Marie, Hege A. Dale, Ragna Sannerud*, and Jaakko Saraste

Department of Biomedicine and Molecular Imaging Center, University of Bergen, N-5009 Bergen, Norway

Submitted December 22, 2008; Revised August 18, 2009; Accepted August 19, 2009
Monitoring Editor: Akihiko Nakano

InCytes from MBC

Because the functional borders of the intermediate compartment (IC) are not well defined, the spatial map of the transport machineries operating between the endoplasmic reticulum (ER) and the Golgi apparatus remains incomplete. Our previous studies showed that the IC consists of interconnected vacuolar and tubular parts with specific roles in pre-Golgi trafficking. Here, using live cell imaging, we demonstrate that the tubules containing the GTPase Rab1A create a long-lived membrane compartment around the centrosome. Separation of this pericentrosomal domain of the IC from the Golgi ribbon, due to centrosome motility, revealed that it contains a distinct pool of COPI coats and acts as a temperature-sensitive way station in post-ER trafficking. However, unlike the Golgi, the pericentrosomal IC resists the disassembly of COPI coats by brefeldin A, maintaining its juxtaposition with the endocytic recycling compartment, and operation as the focal point of a dynamic tubular network that extends to the cell periphery. These results provide novel insight into the compartmental organization of the secretory pathway and Golgi biogenesis. Moreover, they reveal a direct functional connection between the IC and the endosomal system, which evidently contributes to unconventional transport of the cystic fibrosis transmembrane conductance regulator to the cell surface.

This article was published online ahead of print in MBC in Press (http://www.molbiolcell.org/cgi/doi/10.1091/mbc.E08-12-1229) on August 26, 2009.

* Present address: Laboratory for Membrane Trafficking, VIB, Department of Molecular and Developmental Genetics, and Center of Human Genetics, KULeuven, Campus Gasthuisberg, Herestraat 49, Box 602, 3000 Leuven, Belgium.

Address correspondence to: Jaakko E. Saraste (jaakko.saraste{at}biomed.uib.no).

Abbreviations used: BFA, brefeldin A; COP, coat protein; CFTR, cystic fibrosis transmembrane conductance regulator; CM, confocal microscopy; ERC, endocytic recycling compartment; GFP, green fluorescent protein; IC, intermediate compartment; MT, microtubule; pcIC, pericentrosomal domain of the IC; Tf, transferrin.


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