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First published online April 3, 2008
doi: 10.1242/10.1242/jcs.016998

Journal of Cell Science 121, 1293-1302 (2008)
Published by The Company of Biologists 2008
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Research Article

Ypt1p is essential for retrograde Golgi-ER transport and for Golgi maintenance in S. cerevisiae

Faustin Kamena1,*, Melanie Diefenbacher1,2,*, Cornelia Kilchert2, Heinz Schwarz3 and Anne Spang1,2,{dagger}

1 Friedrich Miescher Laboratory of the Max Planck Society, Spemannstrasse 39, 72076 Tübingen, Germany
2 Biozentrum, University of Basel, Klingelbergstrasse 70, CH-4056 Basel, Switzerland
3 Max Planck Institute for Developmental Biology, Spemannstrasse 35, 72076 Tübingen, Germany

{dagger} Author for correspondence (e-mail: anne.spang{at}unibas.ch)

Accepted 28 January 2008

The small GTPase Ypt1p of the Rab family is required for docking of ER-derived transport vesicles with the Golgi prior to fusion. However, the identity of the Rab protein that mediates docking of Golgi-derived COPI vesicles with the ER in retrograde transport remains elusive. Here, we show that in yeast Ypt1p is essential for retrograde transport from the Golgi to the ER. Retrieval of gp{alpha}F-HDEL (glycolylated pro-{alpha}-factor with an HDEL tag at the C-terminus) was blocked in {Delta}ypt1/SLY1-20 membranes at the restrictive temperature in vitro. Moreover, Ypt1p and the ER-resident t-SNARE Ufe1p interact genetically and biochemically, indicating a role for Ypt1p in consumption of COPI vesicles at the ER. Ypt1p is also essential for the maintenance of the morphology and the protein composition of the Golgi. Interestingly, the concentrations of the Golgi enzymes Anp1p and Mnn1p, the cargo protein Emp47p and the v-SNARE Sec22p were all substantially reduced in Golgi from a {Delta}ypt1/SLY1-20 strain as compared with wild-type Golgi, while the concentration of Arf1p and of coatomer were mildly affected. Finally, COPI vesicles generated from {Delta}ypt1/SLY1-20 Golgi membranes in vitro were depleted of Emp47p and Sec22p. These data demonstrate that Ypt1p plays an essential role in retrograde transport from the Golgi to the ER.

Key words: ER-Golgi shuttle, YPT1, Rab, Retrograde transport, Yeast






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