We show here that cytochalasin D-induced depolymerization of actin filaments markedly reduces the stimulus-dependent activation of protein kinase B (PKB) in four different cell types (HEK-293 cells, L6 myotubes, 3T3-L1 adipocytes and U87MG cells). HEK-293 cells expressing the pleckstrin homology (PH) domains of PKB and general receptor for phosphoinositides-1 (GRP1) fused to green fluorescent protein (GFP) were used to monitor production of 3-phosphoinositides in the plasma membrane. Disassembly of the actin cytoskeleton significantly reduced the insulin-mediated translocation of both PKB-PHŐGFP and GRP1-PHŐGFP to the plasma membrane, consistent with diminished synthesis of 3-phosphoinositides. Actin depolymerization did not affect the hormonal activation of phosphoinositide 3-kinase (PI 3-kinase), and since cytochalasin D treatment also led to reduced platelet-derived growth factor (PDGF)-induced phosphorylation of PKB in U87MG cells, a PTEN (phosphatase and tensin homologue deleted on chromosome 10) null cell line, lipid phosphatase activity was unlikely to account for any reduction in cellular 3-phosphoinositides. Withdrawal of cytochalasin D from the extracellular medium induced actin filament repolymerization, and reinstated both the recruitment of PHŐGFP fusion proteins to the plasma membrane and PKB activation in response to insulin and PDGF. Our findings indicate that an intact actin network is a crucial requirement for PI 3-kinase-mediated production of 3-phosphoinositides and, therefore, for the activation of PKB.
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Research Article|
December 08 2000
A role for the actin cytoskeleton in the hormonal and growth-factor-mediated activation of protein kinase B
Karine PEYROLLIER;
Karine PEYROLLIER
1
*School of Life Sciences and the Division of Signal Transduction Therapy1, Medical Sciences Institute/Wellcome Trust Biocentre Complex, Dow Street, University of Dundee, Dundee DD1 5EH, U.K.
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Eric HAJDUCH;
Eric HAJDUCH
1
*School of Life Sciences and the Division of Signal Transduction Therapy1, Medical Sciences Institute/Wellcome Trust Biocentre Complex, Dow Street, University of Dundee, Dundee DD1 5EH, U.K.
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Alexander GRAY;
Alexander GRAY
*School of Life Sciences and the Division of Signal Transduction Therapy1, Medical Sciences Institute/Wellcome Trust Biocentre Complex, Dow Street, University of Dundee, Dundee DD1 5EH, U.K.
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Gary J. LITHERLAND;
Gary J. LITHERLAND
*School of Life Sciences and the Division of Signal Transduction Therapy1, Medical Sciences Institute/Wellcome Trust Biocentre Complex, Dow Street, University of Dundee, Dundee DD1 5EH, U.K.
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Alan R. PRESCOTT;
Alan R. PRESCOTT
*School of Life Sciences and the Division of Signal Transduction Therapy1, Medical Sciences Institute/Wellcome Trust Biocentre Complex, Dow Street, University of Dundee, Dundee DD1 5EH, U.K.
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Nick R. LESLIE;
Nick R. LESLIE
1
*School of Life Sciences and the Division of Signal Transduction Therapy1, Medical Sciences Institute/Wellcome Trust Biocentre Complex, Dow Street, University of Dundee, Dundee DD1 5EH, U.K.
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Harinder S. HUNDAL
Harinder S. HUNDAL
2
*School of Life Sciences and the Division of Signal Transduction Therapy1, Medical Sciences Institute/Wellcome Trust Biocentre Complex, Dow Street, University of Dundee, Dundee DD1 5EH, U.K.
2To whom correspondence should be addressed (e-mail h.s.hundal@dundee.ac.uk).
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Publisher: Portland Press Ltd
Received:
September 26 2000
Revision Received:
October 20 2000
Accepted:
October 30 2000
Online ISSN: 1470-8728
Print ISSN: 0264-6021
The Biochemical Society, London © 2000
2000
Biochem J (2000) 352 (3): 617–622.
Article history
Received:
September 26 2000
Revision Received:
October 20 2000
Accepted:
October 30 2000
Connected Content
A correction has been published:
A role for the actin cytoskeleton in the hormonal and growth-factor-mediated activation of protein kinase B
Citation
Karine PEYROLLIER, Eric HAJDUCH, Alexander GRAY, Gary J. LITHERLAND, Alan R. PRESCOTT, Nick R. LESLIE, Harinder S. HUNDAL; A role for the actin cytoskeleton in the hormonal and growth-factor-mediated activation of protein kinase B. Biochem J 15 December 2000; 352 (3): 617–622. doi: https://doi.org/10.1042/bj3520617
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