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© The Rockefeller University Press, 0021-9525/1999/11/743/ $5.00
The Journal of Cell Biology, Volume 147, Number 4, November 15, 1999 743-760

Original Article

Rab6 Coordinates a Novel Golgi to ER Retrograde Transport Pathway in Live Cells

Jamie Whitea,b, Ludger Johannesc, Frédéric Mallardc, Andreas Girodb, Stephan Grilla,b, Sigrid Reinschb, Patrick Kellerb, Barbara Tzschascheld, Arnaud Echardc, Bruno Goudc, and Ernst H.K. Stelzera,b
a Light Microscopy Group, European Molecular Biology Laboratory, Meyerhofstrasse 1, D-69117 Heidelberg, Germany
b Cell Biophysics and Cell Biology Programme, European Molecular Biology Laboratory, Meyerhofstrasse 1, D-69117 Heidelberg, Germany
c Laboratoire Mécanismes moléculaires du transport intracellulaire, Institut Curie, CNRS UMR 144, Paris, Cedex 05, France
d GBF-National Research Institute for Biotechnology, Department of Microbiology, Mascheroder Weg 1, 38124 Braunschweig, Germany

Correspondence to: Jamie White, Light Microscopy Group and Cell Biophysics and Cell Biology Programme, European Molecular Biology Laboratory, Meyerhofstrasse 1, D-69117 Heidelberg, Germany. Tel:49 6221 387 123 Fax:49 6221 387 306 E-mail:jwhite{at}embl-heidelberg.de.

We visualized a fluorescent-protein (FP) fusion to Rab6, a Golgi-associated GTPase, in conjunction with fluorescent secretory pathway markers. FP-Rab6 defined highly dynamic transport carriers (TCs) translocating from the Golgi to the cell periphery. FP-Rab6 TCs specifically accumulated a retrograde cargo, the wild-type Shiga toxin B-fragment (STB), during STB transport from the Golgi to the endoplasmic reticulum (ER). FP-Rab6 TCs associated intimately with the ER, and STB entered the ER via specialized peripheral regions that accumulated FP-Rab6. Microinjection of antibodies that block coatomer protein I (COPI) function inhibited trafficking of a KDEL-receptor FP-fusion, but not FP-Rab6. Additionally, markers of COPI-dependent recycling were excluded from FP-Rab6/STB TCs. Overexpression of Rab6:GDP (T27N mutant) using T7 vaccinia inhibited toxicity of Shiga holotoxin, but did not alter STB transport to the Golgi or Golgi morphology. Taken together, our results indicate Rab6 regulates a novel Golgi to ER transport pathway.

Key Words: intracellular transport, Shiga toxin, Rab6 protein, KDEL receptor, green fluorescent protein


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