title: Inositol phospholipid 3-kinase is activated by cellular stress but is not required for the stress-induced activation of glucose transport in L6 rat skeletal muscle cells.
abstract: A characteristic response of cells subjected to a stress stimulus is a rapid activation of cellular glucose transport. The mechanisms governing this increase in glucose transport are poorly understood, but it has been suggested that the response may involve the intracellular-signaling components that also participate in the hormonal activation of glucose transport. In skeletal muscle and fat tissue, inositol phospholipid 3-kinase plays an integral role in the regulation of both basal and insulin-stimulated glucose transport. In this study, we have investigated whether inositol phospholipid 3-kinase is activated by chemical stress and, if so, whether it has a role to play in the stress-induced increase in glucose transport in L6 muscle cells. Furthermore, we have attempted to assess the basis by which inositol phospholipid 3-kinase may participate in the regulation of basal glucose transport. Acute exposure (30 min) of L6 muscle cells to 0.5 mM arsenite induced an 80% stimulation in glucose transport. This activation was due to a rise in the number of cell-surface glucose transporters, based on an increase in the Vmax of glucose transport and the observation that arsenite increases the plasma membrane content of GLUT1 and GLUT4 glucose transporters by 95% and 60%, respectively, from an intracellular compartment. Arsenite induced rapid activation (< 2 min) of inositol phospholipid 3-kinase with an approximately fourfold increase in phosphatidylinositol 3,4,5-trisphosphate (PtdIns3,4,5P3). In contrast, phosphatidylinositol 3-phosphate (PtdIns3P) levels were unaffected. Prior treatment of L6 cells with 100 nM wortmannin suppressed the arsenite-induced increase in PtdIns3,4,5P3 and reduced the cellular content of PtdIns3P by 50%. Under these conditions however, wortmannin failed to prevent the stress-induced activation of glucose transport, but suppressed basal glucose transport by 60% with an IC50 of about 10 nM. In the absence of arsenite, wortmannin caused a dose-dependent inhibition in the cellular levels of PtdIns3P and PtdIns3,4,5P3 with IC50 values of about 10 nM and 100 nM, respectively. In summary, the present results demonstrate that chemical stress activates inositol phospholipid 3-kinase and glucose transport in L6 muscle cells, but unlike the hormonal responses of these cells the activation of inositol phospholipid 3-kinase is not responsible for the stress-induced increase in glucose transport. This implies that stress-induced and hormonal stimulated increases in PtdIns3,4,5P3 levels are functionally distinct. By contrast, the maintenance of PtdIns3P levels, presumably involving a PtdIns-specific, wortmannin-sensitive inositol phospholipid 3-kinase may be required to support basal glucose transport.
authors.affiliation: Department of Anatomy and Physiology, The University of Dundee, UK.
authors.forename: H E
authors.initials: HE
authors.lastname: McDowell
authors.forename: T
authors.initials: T
authors.lastname: Walker
authors.forename: E
authors.initials: E
authors.lastname: Hajduch
authors.forename: G
authors.initials: G
authors.lastname: Christie
authors.forename: I H
authors.initials: IH
authors.lastname: Batty
authors.forename: C P
authors.initials: CP
authors.lastname: Downes
authors.forename: H S
authors.initials: HS
authors.lastname: Hundal
mesh_terms: D000730:Androstadienes / Q000494:pharmacology; D000818:Animals; D018053:Arsenites / Q000494:pharmacology*; D002478:Cells, Cultured; D003847:Deoxyglucose / Q000378:metabolism; D004789:Enzyme Activation; D005947:Glucose / Q000378:metabolism*; D051272:Glucose Transporter Type 1; D051275:Glucose Transporter Type 4; D009004:Monosaccharide Transport Proteins / Q000032:analysis; D009124:Muscle Proteins; D018482:Muscle, Skeletal / Q000187:drug effects* / Q000378:metabolism*; D019869:Phosphatidylinositol 3-Kinases; D010716:Phosphatidylinositols / Q000032:analysis; D017853:Phosphotransferases (Alcohol Group Acceptor) / Q000502:physiology*; D051381:Rats; D000077191:Wortmannin
chemical_list: D000730:Androstadienes; D018053:Arsenites; D051272:Glucose Transporter Type 1; D051275:Glucose Transporter Type 4; D009004:Monosaccharide Transport Proteins; D009124:Muscle Proteins; D010716:Phosphatidylinositols; C495679:Slc2a1 protein, rat; C495694:Slc2a4 protein, rat; D003847:Deoxyglucose; D019869:Phosphatidylinositol 3-Kinases; D017853:Phosphotransferases (Alcohol Group Acceptor); D005947:Glucose; C015001:arsenite; D000077191:Wortmannin