Journal
PEERJ
Volume 8, Issue -, Pages -Publisher
PEERJ INC
DOI: 10.7717/peerj.8751
Keywords
Membrane; Transport; Insulin; Diabetes; GLUT4; Endosome
Categories
Funding
- Lord Kelvin/Adam Smith PhD studentship from the University of Glasgow
- Arthur and Sadie Pethybridge studentship from Diabetes UK
- BHF studentship [FS/16/55/32731]
- Diabetes UK [13/0004725, 17/0005605]
- Irish Research Council
- European Research Council through the Consolidator Award FAKIR
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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 GOSRI and YKT6 as potential novel regulators of GLUT4 trafficking in human cells.
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