Morris, S. et al. (2020) Characterisation of GLUT4 trafficking in HeLa cells: comparable kinetics and orthologous trafficking mechanisms to 3T3-L1 adipocytes. PeerJ, 8, e8751. (doi: 10.7717/peerj.8751) (PMID:32185116) (PMCID:PMC7060922)
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Abstract
Insulin-stimulated glucose transport is a characteristic property of adipocytes and muscle cells and involves the regulated delivery of glucose transporter (GLUT4)-containing vesicles from intracellular stores to the cell surface. Fusion of these vesicles results in increased numbers of GLUT4 molecules at the cell surface. In an attempt to overcome some of the limitations associated with both primary and cultured adipocytes, we expressed an epitope- and GFP-tagged version of GLUT4 (HA–GLUT4–GFP) in HeLa cells. Here we report the characterisation of this system compared to 3T3-L1 adipocytes. We show that insulin promotes translocation of HA–GLUT4–GFP to the surface of both cell types with similar kinetics using orthologous trafficking machinery. While the magnitude of the insulin-stimulated translocation of GLUT4 is smaller than mouse 3T3-L1 adipocytes, HeLa cells offer a useful, experimentally tractable, human model system. Here, we exemplify their utility through a small-scale siRNA screen to identify GOSR1 and YKT6 as potential novel regulators of GLUT4 trafficking in human cells.
| Item Type: | Articles |
|---|---|
| Status: | Published |
| Refereed: | Yes |
| Glasgow Author(s) Enlighten ID: | Sadler, Miss Jessica and Black, Hannah Lucy and Koester, Anna and Gould, Professor Gwyn and Cooper, Professor Jonathan and Bryant, Dr Nia and Gadegaard, Professor Nikolaj |
| Authors: | Morris, S., Geoghegan, N. D., Sadler, J. B.A., Koester, A. M., Black, H. L., Laub, M., Miller, L., Heffernan, L., Simpson, J. C., Mastick, C. C., Cooper, J., Gadegaard, N., Bryant, N. J., and Gould, G. W. |
| College/School: | College of Medical Veterinary and Life Sciences > School of Molecular Biosciences College of Science and Engineering > School of Engineering > Biomedical Engineering |
| Journal Name: | PeerJ |
| Publisher: | PeerJ |
| ISSN: | 2167-8359 |
| ISSN (Online): | 2167-8359 |
| Copyright Holders: | Copyright © 2020 Morris et al. |
| First Published: | First published in PeerJ 8: e8751 |
| Publisher Policy: | Reproduced under a Creative Commons License |
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