Characterisation of GLUT4 trafficking in HeLa cells: comparable kinetics and orthologous trafficking mechanisms to 3T3-L1 adipocytes

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:Gould, Professor Gwyn and Sadler, Miss Jessica and Bryant, Dr Nia and Cooper, Professor Jonathan and Koester, Anna and Black, Hannah Lucy 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 > Institute of Molecular Cell and Systems Biology
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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Project CodeAward NoProject NamePrincipal InvestigatorFunder's NameFunder RefLead Dept
173470BHF 4-Year PhD Studentship (4th intake 2016 of 3rd Scheme)Rhian TouyzBritish Heart Foundation (BHF)FS/16/55/32731Institute of Cardiovascular & Medical Sciences
168627Control of GLUT4 sorting by Syntaxin 16 and mVps45 in adipocytesGwyn GouldDiabetes UK (DIABETUK)13/0004725Institute of Molecular, Cell & Systems Biology