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 Filippa, N.
Right arrow Articles by Van Obberghen, E.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Filippa, N.
Right arrow Articles by Van Obberghen, E.

 Previous Article  |  Next Article 

Molecular and Cellular Biology, August 2000, p. 5712-5721, Vol. 20, No. 15
0270-7306/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.

Effect of Phosphoinositide-Dependent Kinase 1 on Protein Kinase B Translocation and Its Subsequent Activation

Nathalie Filippa,1 Carol L. Sable,1 Brian A. Hemmings,2 and Emmanuel Van Obberghen1,*

INSERM U145, IFR 50, Faculté de Médecine, 06107 Nice Cedex 2, France,1 and Friedrich Miescher Institute, CH 4002 Basel, Switzerland2

Received 2 August 1999/Returned for modification 23 September 1999/Accepted 17 April 2000

In this report we investigated the function of phosphoinositide-dependent protein kinase 1 (PDK1) in protein kinase B (PKB) activation and translocation to the cell surface. Wild-type and PDK1 mutants were transfected into HeLa cells, and their subcellular localization was analyzed. PDK1 was found to translocate to the plasma membrane in response to insulin, and this process did not require a functional catalytic activity, since a catalytically inactive kinase mutant (Kd) of PDK1 was capable of translocating. The PDK1 presence at the cell surface was shown to be linked to phospholipids and therefore to serum-dependent phosphatidylinositol 3-kinase activity. Using confocal microscopy in HeLa cells we found that PDK1 colocalizes with PKB at the plasma membrane. Further, after cotransfection of PKB and a PDK1 mutant (Mut) unable to translocate to the plasma membrane, PKB was prevented from moving to the cell periphery after insulin stimulation. In response to insulin, a PKB mutant with its PH domain deleted (Delta PH-PKB) retained the ability to translocate to the plasma membrane when coexpressed with PDK1. Finally, we found that Delta PH-PKB was highly active independent of insulin stimulation when cotransfected with PDK1 mutants defective in their PH domain. These findings suggest that PDK1 brings PKB to the plasma membrane upon exposure of cells to insulin and that the PH domain of PDK1 acts as a negative regulator of its enzyme activity.

* Corresponding author. Mailing address: INSERM U 145, IFR 50, Faculté de Médecine, Avenue de Valombrose, 06107 Nice Cedex 2, France. Phone: 33-4-93-81-54-47. Fax: 33-4-93-81-54-32. E-mail: vanobbeg{at}unice.fr.

Molecular and Cellular Biology, August 2000, p. 5712-5721, Vol. 20, No. 15
0270-7306/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.


This article has been cited by other articles:

  • Restuccia, D. F., Hemmings, B. A. (2009). Blocking Akt-ivity. Science 325: 1083-1084 [Abstract] [Full Text]  
  • Deepa, S. S., Dong, L. Q. (2009). APPL1: role in adiponectin signaling and beyond. Am. J. Physiol. Endocrinol. Metab. 296: E22-E36 [Abstract] [Full Text]  
  • Yang, K.-J., Shin, S., Piao, L., Shin, E., Li, Y., Park, K. A., Byun, H. S., Won, M., Hong, J., Kweon, G. R., Hur, G. M., Seok, J. H., Chun, T., Brazil, D. P., Hemmings, B. A., Park, J. (2008). Regulation of 3-Phosphoinositide-dependent Protein Kinase-1 (PDK1) by Src Involves Tyrosine Phosphorylation of PDK1 and Src Homology 2 Domain Binding. J. Biol. Chem. 283: 1480-1491 [Abstract] [Full Text]  
  • Saito, T., Jones, C. C., Huang, S., Czech, M. P., Pilch, P. F. (2007). The Interaction of Akt with APPL1 Is Required for Insulin-stimulated Glut4 Translocation. J. Biol. Chem. 282: 32280-32287 [Abstract] [Full Text]  
  • Primo, L., di Blasio, L., Roca, C., Droetto, S., Piva, R., Schaffhausen, B., Bussolino, F. (2007). Essential role of PDK1 in regulating endothelial cell migration. JCB 176: 1035-1047 [Abstract] [Full Text]  
  • Riojas, R. A., Kikani, C. K., Wang, C., Mao, X., Zhou, L., Langlais, P. R., Hu, D., Roberts, J. L., Dong, L. Q., Liu, F. (2006). Fine Tuning PDK1 Activity by Phosphorylation at Ser163. J. Biol. Chem. 281: 21588-21593 [Abstract] [Full Text]  
  • Liu, H., Qiu, Y., Xiao, L., Dong, F. (2006). Involvement of Protein Kinase C{epsilon} in the Negative Regulation of Akt Activation Stimulated by Granulocyte Colony-Stimulating Factor. J. Immunol. 176: 2407-2413 [Abstract] [Full Text]  
  • Wu, Y., Zu, K., Warren, M. A., Wallace, P. K., Ip, C. (2006). Delineating the mechanism by which selenium deactivates Akt in prostate cancer cells.. Molecular Cancer Therapeutics 5: 246-252 [Abstract] [Full Text]  
  • Shen, Y. H., Zhang, L., Utama, B., Wang, J., Gan, Y., Wang, X., Wang, J., Chen, L., Vercellotti, G. M., Coselli, J. S., Mehta, J. L., Wang, X. L. (2006). Human cytomegalovirus inhibits Akt-mediated eNOS activation through upregulating PTEN (phosphatase and tensin homolog deleted on chromosome 10). Cardiovasc Res 69: 502-511 [Abstract] [Full Text]  
  • Fiory, F., Alberobello, A. T., Miele, C., Oriente, F., Esposito, I., Corbo, V., Ruvo, M., Tizzano, B., Rasmussen, T. E., Gammeltoft, S., Formisano, P., Beguinot, F. (2005). Tyrosine Phosphorylation of Phosphoinositide-Dependent Kinase 1 by the Insulin Receptor Is Necessary for Insulin Metabolic Signaling. Mol. Cell. Biol. 25: 10803-10814 [Abstract] [Full Text]  
  • Haugaard, S. B., Andersen, O., Madsbad, S., Frosig, C., Iversen, J., Nielsen, J. O., Wojtaszewski, J. F.P. (2005). Skeletal Muscle Insulin Signaling Defects Downstream of Phosphatidylinositol 3-Kinase at the Level of Akt Are Associated With Impaired Nonoxidative Glucose Disposal in HIV Lipodystrophy. Diabetes 54: 3474-3483 [Abstract] [Full Text]  
  • Saxena, S., Bucci, C., Weis, J., Kruttgen, A. (2005). The Small GTPase Rab7 Controls the Endosomal Trafficking and Neuritogenic Signaling of the Nerve Growth Factor Receptor TrkA. J. Neurosci. 25: 10930-10940 [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]  
  • Dong, L. Q., Liu, F. (2005). PDK2: the missing piece in the receptor tyrosine kinase signaling pathway puzzle. Am. J. Physiol. Endocrinol. Metab. 289: E187-E196 [Abstract] [Full Text]  
  • Scheid, M. P., Parsons, M., Woodgett, J. R. (2005). Phosphoinositide-Dependent Phosphorylation of PDK1 Regulates Nuclear Translocation. Mol. Cell. Biol. 25: 2347-2363 [Abstract] [Full Text]  
  • Chandrasekar, B., Mummidi, S., Claycomb, W. C., Mestril, R., Nemer, M. (2005). Interleukin-18 Is a Pro-hypertrophic Cytokine That Acts through a Phosphatidylinositol 3-Kinase-Phosphoinositide-dependent Kinase-1-Akt-GATA4 Signaling Pathway in Cardiomyocytes. J. Biol. Chem. 280: 4553-4567 [Abstract] [Full Text]  
  • Stratford, S., Hoehn, K. L., Liu, F., Summers, S. A. (2004). Regulation of Insulin Action by Ceramide: DUAL MECHANISMS LINKING CERAMIDE ACCUMULATION TO THE INHIBITION OF Akt/PROTEIN KINASE B. J. Biol. Chem. 279: 36608-36615 [Abstract] [Full Text]  
  • Takaya, A., Ohba, Y., Kurokawa, K., Matsuda, M. (2004). RalA Activation at Nascent Lamellipodia of Epidermal Growth Factor-stimulated Cos7 Cells and Migrating Madin-Darby Canine Kidney Cells. Mol. Biol. Cell 15: 2549-2557 [Abstract] [Full Text]  
  • Taniyama, Y., Weber, D. S., Rocic, P., Hilenski, L., Akers, M. L., Park, J., Hemmings, B. A., Alexander, R. W., Griendling, K. K. (2003). Pyk2- and Src-Dependent Tyrosine Phosphorylation of PDK1 Regulates Focal Adhesions. Mol. Cell. Biol. 23: 8019-8029 [Abstract] [Full Text]  
  • Xie, Z., Zeng, X., Waldman, T., Glazer, R. I. (2003). Transformation of Mammary Epithelial Cells by 3-Phosphoinositide- dependent Protein Kinase-1 Activates {beta}-Catenin and c-Myc, and Down-Regulates Caveolin-1. Cancer Res. 63: 5370-5375 [Abstract] [Full Text]  
  • Kim, Y.-B., Kotani, K., Ciaraldi, T. P., Henry, R. R., Kahn, B. B. (2003). Insulin-Stimulated Protein Kinase C {lambda}/{zeta} Activity Is Reduced in Skeletal Muscle of Humans With Obesity and Type 2 Diabetes: Reversal With Weight Reduction. Diabetes 52: 1935-1942 [Abstract] [Full Text]  
  • Ijuin, T., Takenawa, T. (2003). SKIP Negatively Regulates Insulin-Induced GLUT4 Translocation and Membrane Ruffle Formation. Mol. Cell. Biol. 23: 1209-1220 [Abstract] [Full Text]  
  • Smith, L., Smith, J. B. (2002). Lack of Constitutive Activity of the Free Kinase Domain of Protein Kinase C zeta . DEPENDENCE ON TRANSPHOSPHORYLATION OF THE ACTIVATION LOOP. J. Biol. Chem. 277: 45866-45873 [Abstract] [Full Text]  
  • Sato, S., Fujita, N., Tsuruo, T. (2002). Regulation of Kinase Activity of 3-Phosphoinositide-dependent Protein Kinase-1 by Binding to 14-3-3. J. Biol. Chem. 277: 39360-39367 [Abstract] [Full Text]  
  • Conus, N. M., Hannan, K. M., Cristiano, B. E., Hemmings, B. A., Pearson, R. B. (2002). Direct Identification of Tyrosine 474 as a Regulatory Phosphorylation Site for the Akt Protein Kinase. J. Biol. Chem. 277: 38021-38028 [Abstract] [Full Text]  
  • Scheid, M. P., Marignani, P. A., Woodgett, J. R. (2002). Multiple Phosphoinositide 3-Kinase-Dependent Steps in Activation of Protein Kinase B. Mol. Cell. Biol. 22: 6247-6260 [Abstract] [Full Text]  
  • Kandasamy, K., Srivastava, R. K. (2002). Role of the Phosphatidylinositol 3'-Kinase/PTEN/Akt Kinase Pathway in Tumor Necrosis Factor-related Apoptosis-inducing Ligand-induced Apoptosis in Non-Small Cell Lung Cancer Cells. Cancer Res. 62: 4929-4937 [Abstract] [Full Text]  
  • Montagnani, M., Ravichandran, L. V., Chen, H., Esposito, D. L., Quon, M. J. (2002). Insulin Receptor Substrate-1 and Phosphoinositide-Dependent Kinase-1 Are Required for Insulin-Stimulated Production of Nitric Oxide in Endothelial Cells. Mol. Endocrinol. 16: 1931-1942 [Abstract] [Full Text]  
  • Wick, M. J., Wick, K. R., Chen, H., He, H., Dong, L. Q., Quon, M. J., Liu, F. (2002). Substitution of the Autophosphorylation Site Thr516 with a Negatively Charged Residue Confers Constitutive Activity to Mouse 3-Phosphoinositide-dependent Protein Kinase-1 in Cells. J. Biol. Chem. 277: 16632-16638 [Abstract] [Full Text]  
  • Bourbon, N. A., Sandirasegarane, L., Kester, M. (2002). Ceramide-induced Inhibition of Akt Is Mediated through Protein Kinase Czeta . IMPLICATIONS FOR GROWTH ARREST. J. Biol. Chem. 277: 3286-3292 [Abstract] [Full Text]  
  • Thakkar, H., Chen, X., Tyan, F., Gim, S., Robinson, H., Lee, C., Pandey, S. K., Nwokorie, C., Onwudiwe, N., Srivastava, R. K. (2001). Pro-survival Function of Akt/Protein Kinase B in Prostate Cancer Cells. RELATIONSHIP WITH TRAIL RESISTANCE. J. Biol. Chem. 276: 38361-38369 [Abstract] [Full Text]  
  • Cantrell, D. (2001). Phosphoinositide 3-kinase signalling pathways. J. Cell Sci. 114: 1439-1445 [Abstract]  
  • Wick, M. J., Dong, L. Q., Riojas, R. A., Ramos, F. J., Liu, F. (2000). Mechanism of Phosphorylation of Protein Kinase B/Akt by a Constitutively Active 3-Phosphoinositide-dependent Protein Kinase-1. J. Biol. Chem. 275: 40400-40406 [Abstract] [Full Text]  
  • Shigematsu, S., Miller, S. L., Pessin, J. E. (2001). Differentiated 3T3L1 Adipocytes Are Composed of Heterogenous Cell Populations with Distinct Receptor Tyrosine Kinase Signaling Properties. J. Biol. Chem. 276: 15292-15297 [Abstract] [Full Text]  
  • Ballif, B. A., Shimamura, A., Pae, E., Blenis, J. (2001). Disruption of 3-Phosphoinositide-dependent Kinase 1 (PDK1) Signaling by the Anti-tumorigenic and Anti-proliferative Agent N-alpha -tosyl-L-phenylalanyl Chloromethyl Ketone. J. Biol. Chem. 276: 12466-12475 [Abstract] [Full Text]  
  • Jacobs, A. R., LeRoith, D., Taylor, S. I. (2001). Insulin Receptor Substrate-1 Pleckstrin Homology and Phosphotyrosine-binding Domains Are Both Involved in Plasma Membrane Targeting. J. Biol. Chem. 276: 40795-40802 [Abstract] [Full Text]  
  • Park, J., Hill, M. M., Hess, D., Brazil, D. P., Hofsteenge, J., Hemmings, B. A. (2001). Identification of Tyrosine Phosphorylation Sites on 3-Phosphoinositide-dependent Protein Kinase-1 and Their Role in Regulating Kinase Activity. J. Biol. Chem. 276: 37459-37471 [Abstract] [Full Text]  
  • Sonnenburg, E. D., Gao, T., Newton, A. C. (2001). The Phosphoinositide-dependent Kinase, PDK-1, Phosphorylates Conventional Protein Kinase C Isozymes by a Mechanism That Is Independent of Phosphoinositide 3-Kinase. J. Biol. Chem. 276: 45289-45297 [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»