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Originally published as MBC in Press, 10.1091/mbc.E02-06-0315 on December 7, 2002
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Vol. 14, Issue 3, 973-986, March 2003

GLUT4 Recycles via a trans-Golgi Network (TGN) Subdomain Enriched in Syntaxins 6 and 16 But Not TGN38: Involvement of an Acidic Targeting Motif

Annette M. Shewan,*dagger Dagger Ellen M. van Dam,*§ Sally Martin,dagger Dagger Tang Bor Luen,|| Wanjin Hong,|| Nia J. Bryant,§ and David E. James§

 dagger Institute for Molecular Biosciences and  Dagger Department of Physiology and Pharmacology, University of Queensland, St. Lucia, Brisbane, QLD 4072, Australia;  §Garvan Institute of Medical Research, St. Vincent's Hospital, Darlinghurst, 2010 New South Wales, Australia; and  ||Membrane Biology Laboratory, Institute of Molecular and Cell Biology, National University of Singapore, Singapore 117609

Insulin stimulates glucose transport in fat and muscle cells by triggering exocytosis of the glucose transporter GLUT4. To define the intracellular trafficking of GLUT4, we have studied the internalization of an epitope-tagged version of GLUT4 from the cell surface. GLUT4 rapidly traversed the endosomal system en route to a perinuclear location. This perinuclear GLUT4 compartment did not colocalize with endosomal markers (endosomal antigen 1 protein, transferrin) or TGN38, but showed significant overlap with the TGN target (t)-soluble N-ethylmaleimide-sensitive factor attachment protein receptors (SNAREs) Syntaxins 6 and 16. These results were confirmed by vesicle immunoisolation. Consistent with a role for Syntaxins 6 and 16 in GLUT4 trafficking we found that their expression was up-regulated significantly during adipocyte differentiation and insulin stimulated their movement to the cell surface. GLUT4 trafficking between endosomes and trans-Golgi network was regulated via an acidic targeting motif in the carboxy terminus of GLUT4, because a mutant lacking this motif was retained in endosomes. We conclude that GLUT4 is rapidly transported from the cell surface to a subdomain of the trans-Golgi network that is enriched in the t-SNAREs Syntaxins 6 and 16 and that an acidic targeting motif in the C-terminal tail of GLUT4 plays an important role in this process.


* These authors contributed equally to this work.

Corresponding author. E-mail address: d.james{at}garvan.org.au.


Molecular Biology of the Cell
Vol. 14, 973-986, March 2003
Copyright © 2003 by The American Society for Cell Biology



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