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

Autor:	Morris S; Institute of Molecular Cell and Systems Biology, University of Glasgow, Glasgow, UK., Geoghegan ND; School of Engineering, University of Glasgow, Glasgow, UK., Sadler JBA; Institute of Molecular Cell and Systems Biology, University of Glasgow, Glasgow, UK., Koester AM; Institute of Molecular Cell and Systems Biology, University of Glasgow, Glasgow, UK., Black HL; Department of Biology, University of York, York, UK., Laub M; Institute of Molecular Cell and Systems Biology, University of Glasgow, Glasgow, UK., Miller L; Institute of Molecular Cell and Systems Biology, University of Glasgow, Glasgow, UK., Heffernan L; School of Biology & Environmental Science, University College Dublin, Dublin, Ireland., Simpson JC; School of Biology & Environmental Science, University College Dublin, Dublin, Ireland., Mastick CC; Molecular Biosciences, University of Nevada - Reno, Reno, NV, USA., Cooper J; School of Engineering, University of Glasgow, Glasgow, UK., Gadegaard N; School of Engineering, University of Glasgow, Glasgow, UK., Bryant NJ; Department of Biology, University of York, York, UK., Gould GW; Strathclyde Institute of Pharmacy and Biomedical Sciences, University of Strathclyde, Glasgow, UK.
Jazyk:	angličtina
Zdroj:	PeerJ [PeerJ] 2020 Mar 05; Vol. 8, pp. e8751. Date of Electronic Publication: 2020 Mar 05 (Print Publication: 2020).
DOI:	10.7717/peerj.8751
Abstrakt:	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. Competing Interests: Gwyn W. Gould is an Academic Editor for PeerJ. (© 2020 Morris et al.)
Databáze:	MEDLINE
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