Exofacial epitope-tagged glucose transporter chimeras reveal COOH- terminal sequences governing cellular localization

doi:10.1083/jcb.123.1.127

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Exofacial epitope-tagged glucose transporter chimeras reveal COOH- terminal sequences governing cellular localization

MP Czech, A Chawla, CW Woon, J Buxton, M Armoni, W Tang, M Joly and S Corvera
Program in Molecular Medicine, University of Massachusetts Medical Center, Worcester 01605.

The insulin-regulated adipocyte/skeletal muscle glucose transporter (GLUT4) displays a characteristic steady-state intracellular localization under basal conditions, whereas the erythrocyte/brain transporter isoform (GLUT1) distributes mostly to the cell surface. To identify possible structural elements in these transporter proteins that determine their cellular localization, GLUT1/GLUT4 chimera cDNA constructs that contain the hemagglutinin epitope YPYDVPDYA (HA) in their major exofacial loops were engineered. Binding of monoclonal anti- HA antibody to non-permeabilized COS-7 cells expressing HA-tagged transporter chimeras revealed that expression of transporters on the cell surface was strongly influenced by their cytoplasmic COOH-terminal domain. This method also revealed a less marked, but significant effect on cellular localization of amino acid residues between transporter exofacial and middle loops. The subcellular distribution of expressed chimeras was confirmed by immunofluorescence microscopy of permeabilized COS-7 cells. Thus, HA-tagged native GLUT4 was concentrated in the perinuclear region, whereas a chimera containing the COOH-terminal 29 residues of GLUT1 substituted onto GLUT4 distributed to the plasma membrane, as did native GLUT1. Furthermore, a chimera composed of GLUT1 with a GLUT4 COOH-terminal 30-residue substitution exhibited a predominantly intracellular localization. Similar data was obtained in CHO cells stably expressing these chimeras. Taken together, these results define the unique COOH-terminal cytoplasmic sequences of the GLUT1 and GLUT4 glucose transporters as important determinants of cellular localization in COS-7 and CHO cells.
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Katagiri, H., Asano, T., Yamada, T., Aoyama, T., Fukushima, Y., Kikuchi, M., Kodama, T., Oka, Y. (2002). Acyl-Coenzyme A Dehydrogenases Are Localized on GLUT4-Containing Vesicles via Association with Insulin-Regulated Aminopeptidase in a Manner Dependent on Its Dileucine Motif. Mol. Endocrinol. 16: 1049-1059 [Abstract] [Full Text]
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Kao, A. W., Noda, Y., Johnson, J. H., Pessin, J. E., Saltiel, A. R. (1999). Aldolase Mediates the Association of F-actin with the Insulin-responsive Glucose Transporter GLUT4. J. Biol. Chem. 274: 17742-17747 [Abstract] [Full Text]
Wang, Q., Somwar, R., Bilan, P. J., Liu, Z., Jin, J., Woodgett, J. R., Klip, A. (1999). Protein Kinase B/Akt Participates in GLUT4 Translocation by Insulin in L6 Myoblasts. Mol. Cell. Biol. 19: 4008-4018 [Abstract] [Full Text]
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Kao, A. W., Ceresa, B. P., Santeler, S. R., Pessin, J. E. (1998). Expression of a Dominant Interfering Dynamin Mutant in 3T3L1 Adipocytes Inhibits GLUT4 Endocytosis without Affecting Insulin Signaling. J. Biol. Chem. 273: 25450-25457 [Abstract] [Full Text]
Frevert, E. U., Bjorbak, C., Venable, C. L., Keller, S. R., Kahn, B. B. (1998). Targeting of Constitutively Active Phosphoinositide 3-Kinase to GLUT4-containing Vesicles in 3T3-L1 Adipocytes. J. Biol. Chem. 273: 25480-25487 [Abstract] [Full Text]
Johnson, A. O., Subtil, A., Petrush, R., Kobylarz, K., Keller, S. R., McGraw, T. E. (1998). Identification of an Insulin-responsive, Slow Endocytic Recycling Mechanism in Chinese Hamster Ovary Cells. J. Biol. Chem. 273: 17968-17977 [Abstract] [Full Text]
Suva, L. J., Flannery, M. S., Caulfield, M. P., Findlay, D. M., Jüppner, H., Goldring, S. R., Rosenblatt, M., Chorev, M. (1997). Design, Synthesis and Utility of Novel Benzophenone-Containing Calcitonin Analogs for Photoaffinity Labeling the Calcitonin Receptor. J. Pharmacol. Exp. Ther. 283: 876-884 [Abstract] [Full Text]
Waters, S. B., D'Auria, M., Martin, S. S., Nguyen, C., Kozma, L. M., Luskey, K. L. (1997). The Amino Terminus of Insulin-responsive Aminopeptidase Causes Glut4 Translocation in 3T3-L1 Adipocytes. J. Biol. Chem. 272: 23323-23327 [Abstract] [Full Text]
Lee, W., Jung, C. Y. (1997). A Synthetic Peptide Corresponding to the GLUT4 C-terminal Cytoplasmic Domain Causes Insulin-like Glucose Transport Stimulation and GLUT4 Recruitment in Rat Adipocytes. J. Biol. Chem. 272: 21427-21431 [Abstract] [Full Text]
Garippa, R. J., Johnson, A., Park, J., Petrush, R. L., McGraw, T. E. (1996). The Carboxyl Terminus of GLUT4 Contains a Serine-Leucine-Leucine Sequence That Functions as a Potent Internalization Motif in Chinese Hamster Ovary Cells. J. Biol. Chem. 271: 20660-20668 [Abstract] [Full Text]
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Tozzo, E., Kahn, B. B., Pilch, P. F., Kandror, K. V. (1996). Glut4 Is Targeted to Specific Vesicles in Adipocytes of Transgenic Mice Overexpressing Glut4 Selectively in Adipose Tissue. J. Biol. Chem. 271: 10490-10494 [Abstract] [Full Text]
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