Live imaging of bidirectional traffic from the ERGIC

J Cell Sci. 2005 Jan 15;118(Pt 2):357-67. doi: 10.1242/jcs.01615. Epub 2005 Jan 4.

Abstract

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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Biological Transport, Active / physiology
  • Endoplasmic Reticulum / metabolism
  • Golgi Apparatus / metabolism*
  • Green Fluorescent Proteins / metabolism
  • HeLa Cells
  • Humans
  • Mannose-Binding Lectins / metabolism*
  • Membrane Proteins / metabolism*
  • Protein Transport / physiology

Substances

  • LMAN1 protein, human
  • Mannose-Binding Lectins
  • Membrane Proteins
  • Green Fluorescent Proteins