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MBC in Press, published online ahead of print June 23, 2004
Mol. Biol. Cell 10.1091/mbc.E03-12-0876

A more recent version of this article appeared on September 1, 2004
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Submitted on December 9, 2003
Revised on May 12, 2004
Accepted on May 13, 2004

Participation of the Syntaxin 5/Ykt6/GS28/GS15 SNARE Complex in Transport from the Early/Recycling Endosome to the TGN

Guihua Tai*, Lei Lu*, Tuan Lao Wang*, Bor Luen Tang{dagger}, Bruno Goud{ddagger}, Ludger Johannes{sect}, and Wanjin Hong*||

*Membrane Biology Laboratory, Institute of Molecular and Cell Biology, Singapore 117609, Singapore; {dagger}Department of Biochemistry and Neurobiology Program, National University of Singapore, Singapore 117597, Singapore; and {ddagger}Molecular Mechanisms of Intracellular Transport and {sect}Traffic and Signaling Laboratories, UMR 144 Curie/CNRS, Institut Curie, F-75248 Paris Cedex 05, France

Monitoring Editor: Jean Gruenberg

An in vitro transport assay, established with a modified Shiga toxin B subunit as a marker, has proved to be useful for the study of transport from the early/recycling endosome (EE/RE) to the TGN. Here, we modified this assay to test antibodies to all known SNAREs that have been shown to localize in the Golgi, and found that syntaxin 5, GS28, Ykt6 and GS15 antibodies specifically inhibited STxB transport. Since syntaxin 5, GS28, Ykt6, and GS15 exist as a unique SNARE complex, our observation indicates that these four SNAREs function as a complex in EE/RE-TGN transport. The importance of GS15 in EE/RE-TGN transport was further demonstrated by a block in STxB transport in HeLa cells when GS15 expression was knocked down by its siRNA. Morphological analyses showed that some GS15 and Ykt6 were redistributed from the Golgi to the endosomes when the recycling endosome was perturbed by SNX3-overexpression, suggesting that GS15 and Ykt6 might cycle between the endosomes and the Golgi apparatus. Further studies indicated that syntaxin 5 and syntaxin 16 exerted their role in EE/RE-TGN transport in an additive manner. The kinetics of inhibition exhibited by syntaxin 16 and syntaxin 5 antibodies is similar.

||Corresponding author. E-mail: mcbhwj{at}imcb.a-star.edu.sg




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