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First published online 4 January 2005
doi: 10.1242/jcs.01615

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Live imaging of bidirectional traffic from the ERGIC

¹ Department of Pharmacology and Neurobiology, Biozentrum, University of Basel, Klingelbergstrasse 70, 4056 Basel, Switzerland
² Department of Cell Biology and Biophysics, European Molecular Biology Laboratory Heidelberg, 69117 Heidelberg, Germany

^* Author for correspondence (e-mail: hans-peter.hauri{at}unibas.ch)

Accepted 27 October 2004

The endoplasmic reticulum-Golgi intermediate compartment (ERGIC) defined by the cycling lectin ERGIC-53 consists of tubulovesicular clusters, but it is unknown if these membranes are transport vehicles or stationary entities. Here, we show by live imaging that GFP-ERGIC-53 mainly localizes to long-lived stationary and some short-lived highly mobile elements. Unlike the anterograde marker VSV-G-GFP, GFP-ERGIC-53 does not vectorially move to the Golgi upon exit from the ERGIC, as assessed by a novel quantitative vector field method. Dual-color imaging of GFP-ERGIC-53 and a secretory protein (signal-sequence-tagged dsRed) reveals that the stationary elements are sites of repeated sorting of retrograde and anterograde cargo, and are interconnected by highly mobile elements. These results suggest that the ERGIC is stationary and not simply a collection of mobile carriers that mediate protein traffic from endoplasmic reticulum to Golgi.

Key words: ERGIC-53, Live cell imaging, Membrane traffic, Secretory pathway, Vesicular stomatitis virus G protein

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