This Article
Right arrow Full Text
Right arrow Full Text (PDF)
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Services
Right arrow Similar articles in this journal
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Download to citation manager
Right arrowReprints and Permissions
Right arrow Copyright Information
Right arrow Books from ASM Press
Right arrow MicrobeWorld
Citing Articles
Right arrow Citing Articles via HighWire
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by Patel, N.
Right arrow Articles by Klip, A.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Patel, N.
Right arrow Articles by Klip, A.

 Previous Article  |  Next Article 

Molecular and Cellular Biology, July 2003, p. 4611-4626, Vol. 23, No. 13
0270-7306/03/$08.00+0     DOI: 10.1128/MCB.23.13.4611-4626.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.

Intracellular Segregation of Phosphatidylinositol-3,4,5-Trisphosphate by Insulin-Dependent Actin Remodeling in L6 Skeletal Muscle Cells

Nish Patel,1,2 Assaf Rudich,1 Zayna A. Khayat,1 Rami Garg,1 and Amira Klip1,2*

Programme in Cell Biology, The Hospital for Sick Children, Toronto, Ontario, Canada M5G 1X8,1 Department of Physiology, University of Toronto, Toronto, Ontario, Canada M5S 1A82

Received 20 August 2002/ Returned for modification 12 November 2002/ Accepted 16 April 2003

Insulin stimulates glucose uptake by recruiting glucose transporter 4 (GLUT4) from an intracellular pool to the cell surface through a mechanism that is dependent on phosphatidylinositol (PI) 3-kinase (PI3-K) and cortical actin remodeling. Here we test the hypothesis that insulin-dependent actin filament remodeling determines the location of insulin signaling molecules. It has been shown previously that insulin treatment of L6 myotubes leads to a rapid rearrangement of actin filaments into submembrane structures where the p85 regulatory subunit of PI3-K and organelles containing GLUT4, VAMP2, and the insulin-regulated aminopeptidase (IRAP) colocalize. We now report that insulin receptor substrate-1 and the p110{alpha} catalytic subunit of PI3-K (but not p110ß) also colocalize with the actin structures. Akt-1 was also found in the remodeled actin structures, unlike another PI3-K effector, atypical protein kinase C {lambda}. Transiently transfected green fluorescent protein (GFP)-tagged pleckstrin homology (PH) domains of general receptor for phosphoinositides-1 (GRP1) or Akt (ligands of phosphatidylinositol-3,4,5-trisphosphate [PI-3,4,5-P3]) migrated to the periphery of the live cells; in fixed cells, they were detected in the insulin-induced actin structures. These results suggest that PI-3,4,5-P3 is generated on membranes located within the actin mesh. Actin remodeling and GLUT4 externalization were blocked in cells highly expressing GFP-PH-GRP1, suggesting that PI-3,4,5-P3 is required for both phenomena. We propose that PI-3,4,5-P3 leads to actin remodeling, which in turn segregates p85{alpha} and p110{alpha}, thus localizing PI-3,4,5-P3 production on membranes trapped by the actin mesh. Insulin-stimulated actin remodeling may spatially coordinate the localized generation of PI-3,4,5-P3 and recruitment of Akt, ultimately leading to GLUT4 insertion at the plasma membrane.

* Corresponding author. Mailing address: Programme in Cell Biology, Hospital for Sick Children, 555 University Ave., Toronto, Ontario, Canada M5G 1X8. Phone: (416) 813-6392. Fax: (416) 813-5028. E-mail: amira{at}sickkids.ca.

Molecular and Cellular Biology, July 2003, p. 4611-4626, Vol. 23, No. 13
0022-538X/03/$08.00+0     DOI: 10.1128/MCB.23.13.4611-4626.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.



This article has been cited by other articles:

  • Kachko, I., Maissel, A., Mazor, L., Ben-Romano, R., Watson, R. T., Hou, J. C., Pessin, J. E., Bashan, N., Rudich, A. (2009). Postreceptoral Adipocyte Insulin Resistance Induced by Nelfinavir Is Caused by Insensitivity of PKB/Akt to Phosphatidylinositol-3,4,5-Trisphosphate. Endocrinology 150: 2618-2626 [Abstract] [Full Text]  
  • Bashan, N., Kovsan, J., Kachko, I., Ovadia, H., Rudich, A. (2009). Positive and Negative Regulation of Insulin Signaling by Reactive Oxygen and Nitrogen Species. Physiol. Rev. 89: 27-71 [Abstract] [Full Text]  
  • Carranza, A., Musolino, P. L., Villar, M., Nowicki, S. (2008). Signaling cascade of insulin-induced stimulation of L-dopa uptake in renal proximal tubule cells. Am. J. Physiol. Cell Physiol. 295: C1602-C1609 [Abstract] [Full Text]  
  • Randhawa, V. K., Ishikura, S., Talior-Volodarsky, I., Cheng, A. W. P., Patel, N., Hartwig, J. H., Klip, A. (2008). GLUT4 Vesicle Recruitment and Fusion Are Differentially Regulated by Rac, AS160, and Rab8A in Muscle Cells. J. Biol. Chem. 283: 27208-27219 [Abstract] [Full Text]  
  • Talior-Volodarsky, I., Randhawa, V. K., Zaid, H., Klip, A. (2008). {alpha}-Actinin-4 Is Selectively Required for Insulin-induced GLUT4 Translocation. J. Biol. Chem. 283: 25115-25123 [Abstract] [Full Text]  
  • Lindsay, Y., McCoull, D., Davidson, L., Leslie, N. R., Fairservice, A., Gray, A., Lucocq, J., Downes, C. P. (2006). Localization of agonist-sensitive PtdIns(3,4,5)P3 reveals a nuclear pool that is insensitive to PTEN expression. J. Cell Sci. 119: 5160-5168 [Abstract] [Full Text]  
  • Hou, J. C., Suzuki, N., Pessin, J. E., Watson, R. T. (2006). A Specific Dileucine Motif Is Required for the GGA-dependent Entry of Newly Synthesized Insulin-responsive Aminopeptidase into the Insulin-responsive Compartment. J. Biol. Chem. 281: 33457-33466 [Abstract] [Full Text]  
  • Wijesekara, N., Tung, A., Thong, F., Klip, A. (2006). Muscle cell depolarization induces a gain in surface GLUT4 via reduced endocytosis independently of AMPK. Am. J. Physiol. Endocrinol. Metab. 290: E1276-E1286 [Abstract] [Full Text]  
  • Ishiki, M., Klip, A. (2005). Minireview: Recent Developments in the Regulation of Glucose Transporter-4 Traffic: New Signals, Locations, and Partners. Endocrinology 146: 5071-5078 [Abstract] [Full Text]  
  • Ishiki, M., Randhawa, V. K., Poon, V., JeBailey, L., Klip, A. (2005). Insulin Regulates the Membrane Arrival, Fusion, and C-terminal Unmasking of Glucose Transporter-4 via Distinct Phosphoinositides. J. Biol. Chem. 280: 28792-28802 [Abstract] [Full Text]  
  • Thong, F. S. L., Dugani, C. B., Klip, A. (2005). Turning Signals On and Off: GLUT4 Traffic in the Insulin-Signaling Highway. Physiology 20: 271-284 [Abstract] [Full Text]  
  • Cui, X.-L., Schlesier, A. M., Fisher, E. L., Cerqueira, C., Ferraris, R. P. (2005). Fructose-induced increases in neonatal rat intestinal fructose transport involve the PI3-kinase/Akt signaling pathway. Am. J. Physiol. Gastrointest. Liver Physiol. 288: G1310-G1320 [Abstract] [Full Text]  
  • Huang, C., Thirone, A. C. P., Huang, X., Klip, A. (2005). Differential Contribution of Insulin Receptor Substrates 1 Versus 2 to Insulin Signaling and Glucose Uptake in L6 Myotubes. J. Biol. Chem. 280: 19426-19435 [Abstract] [Full Text]  
  • Eyster, C. A., Duggins, Q. S., Olson, A. L. (2005). Expression of Constitutively Active Akt/Protein Kinase B Signals GLUT4 Translocation in the Absence of an Intact Actin Cytoskeleton. J. Biol. Chem. 280: 17978-17985 [Abstract] [Full Text]  
  • Torok, D., Patel, N., JeBailey, L., Thong, F. S. L., Randhawa, V. K., Klip, A., Rudich, A. (2004). Insulin but not PDGF relies on actin remodeling and on VAMP2 for GLUT4 translocation in myoblasts. J. Cell Sci. 117: 5447-5455 [Abstract] [Full Text]  
  • Brozinick, J. T. Jr., Hawkins, E. D., Strawbridge, A. B., Elmendorf, J. S. (2004). Disruption of Cortical Actin in Skeletal Muscle Demonstrates an Essential Role of the Cytoskeleton in Glucose Transporter 4 Translocation in Insulin-sensitive Tissues. J. Biol. Chem. 279: 40699-40706 [Abstract] [Full Text]  
  • Sweeney, G., Garg, R. R., Ceddia, R. B., Li, D., Ishiki, M., Somwar, R., Foster, L. J., Neilsen, P. O., Prestwich, G. D., Rudich, A., Klip, A. (2004). Intracellular Delivery of Phosphatidylinositol (3,4,5)-Trisphosphate Causes Incorporation of Glucose Transporter 4 into the Plasma Membrane of Muscle and Fat Cells without Increasing Glucose Uptake. J. Biol. Chem. 279: 32233-32242 [Abstract] [Full Text]  
  • Bose, A., Robida, S., Furcinitti, P. S., Chawla, A., Fogarty, K., Corvera, S., Czech, M. P. (2004). Unconventional Myosin Myo1c Promotes Membrane Fusion in a Regulated Exocytic Pathway. Mol. Cell. Biol. 24: 5447-5458 [Abstract] [Full Text]  
  • JeBailey, L., Rudich, A., Huang, X., Ciano-Oliveira, C. D., Kapus, A., Klip, A. (2004). Skeletal Muscle Cells and Adipocytes Differ in Their Reliance on TC10 and Rac for Insulin-Induced Actin Remodeling. Mol. Endocrinol. 18: 359-372 [Abstract] [Full Text]  


Copyright © 2009 by the American Society for Microbiology.   For an alternate route to Journals.ASM.org, visit: http://intl-journals.asm.org | More Info»