This Article
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 McIlroy, J.
Right arrow Articles by Backer, J. M.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by McIlroy, J.
Right arrow Articles by Backer, J. M.

 Previous Article  |  Next Article 

Mol. Cell. Biol., 01 1997, 248-255, Vol 17, No. 1
Copyright © 1997, American Society for Microbiology

Specific activation of p85-p110 phosphatidylinositol 3'-kinase stimulates DNA synthesis by ras- and p70 S6 kinase-dependent pathways

J McIlroy, D Chen, C Wjasow, T Michaeli and JM Backer
Department of Molecular Pharmacology, Albert Einstein College of Medicine, Bronx, New York 10461, USA.

We have developed a polyclonal antibody that activates the heterodimeric p85-p110 phosphatidylinositol (PI) 3'-kinase in vitro and in microinjected cells. Affinity purification revealed that the activating antibody recognized the N-terminal SH2 (NSH2) domain of p85, and the antibody increased the catalytic activity of recombinant p85- p110 dimers threefold in vitro. To study the role of endogenous PI 3'- kinase in intact cells, the activating anti-NSH2 antibody was microinjected into GRC + LR73 cells, a CHO cell derivative selected for tight quiescence during serum withdrawal. Microinjection of anti-NSH2 antibodies increased bromodeoxyuridine (BrdU) incorporation fivefold in quiescent cells and enhanced the response to serum. These data reflect a specific activation of PI 3'-kinase, as the effect was blocked by coinjection of the appropriate antigen (glutathione S-transferase-NSH2 domains from p85 alpha), coinjection of inhibitory anti-p110 antibodies, or treatment of cells with wortmannin. We used the activating antibodies to study signals downstream from PI 3'-kinase. Although treatment of cells with 50 nM rapamycin only partially decreased anti-NSH2-stimulated BrdU incorporation, coinjection with an anti-p70 S6 kinase antibody effectively blocked anti-NSH2-stimulated DNA synthesis. We also found that coinjection of inhibitory anti-ras antibodies blocked both serum- and anti-NSH2-stimulated BrdU incorporation by approximately 60%, and treatment of cells with a specific inhibitor of MEK abolished antibody-stimulated BrdU incorporation. We conclude that selective activation of physiological levels of PI 3'-kinase is sufficient to stimulate DNA synthesis in quiescent cells. PI 3'-kinase-mediated DNA synthesis requires both p70 S6 kinase and the P21ras/MEK pathway.


This article has been cited by other articles:

  • Thimmaiah, K. N., Easton, J. B., Germain, G. S., Morton, C. L., Kamath, S., Buolamwini, J. K., Houghton, P. J. (2005). Identification of N10-Substituted Phenoxazines as Potent and Specific Inhibitors of Akt Signaling. J. Biol. Chem. 280: 31924-31935 [Abstract] [Full Text]  
  • Sawyer, C., Sturge, J., Bennett, D. C., O'Hare, M. J., Allen, W. E., Bain, J., Jones, G. E., Vanhaesebroeck, B. (2003). Regulation of Breast Cancer Cell Chemotaxis by the Phosphoinositide 3-Kinase p110{delta}. Cancer Res. 63: 1667-1675 [Abstract] [Full Text]  
  • Meuillet, E. J., Mahadevan, D., Vankayalapati, H., Berggren, M., Williams, R., Coon, A., Kozikowski, A. P., Powis, G. (2003). Specific Inhibition of the Akt1 Pleckstrin Homology Domain by D-3-Deoxy-Phosphatidyl-myo-Inositol Analogues. Molecular Cancer Therapeutics 2: 389-399 [Abstract] [Full Text]  
  • Strair, R. K., Schaar, D., Goodell, L., Aisner, J., Chin, K.-V., Eid, J., Senzon, R., Cui, X. X., Han, Z. T., Knox, B., Rabson, A. B., Chang, R., Conney, A. (2002). Administration of a Phorbol Ester to Patients with Hematological Malignancies: Preliminary Results from a Phase I Clinical Trial of 12-O-Tetradecanoylphorbol-13-acetate. Clin. Cancer Res. 8: 2512-2518 [Abstract] [Full Text]  
  • Goncharova, E. A., Ammit, A. J., Irani, C., Carroll, R. G., Eszterhas, A. J., Panettieri, R. A., Krymskaya, V. P. (2002). PI3K is required for proliferation and migration of human pulmonary vascular smooth muscle cells. Am. J. Physiol. Lung Cell. Mol. Physiol. 283: L354-L363 [Abstract] [Full Text]  
  • Kops, G. J. P. L., Medema, R. H., Glassford, J., Essers, M. A. G., Dijkers, P. F., Coffer, P. J., Lam, E. W.-F., Burgering, B. M. T. (2002). Control of Cell Cycle Exit and Entry by Protein Kinase B-Regulated Forkhead Transcription Factors. Mol. Cell. Biol. 22: 2025-2036 [Abstract] [Full Text]  
  • Li, B.-S., Ma, W., Zhang, L., Barker, J. L., Stenger, D. A., Pant, H. C. (2001). Activation of Phosphatidylinositol-3 Kinase (PI-3K) and Extracellular Regulated Kinases (Erk1/2) Is Involved in Muscarinic Receptor-Mediated DNA Synthesis in Neural Progenitor Cells. J. Neurosci. 21: 1569-1579 [Abstract] [Full Text]  
  • Montcouquiol, M., Corwin, J. T. (2001). Intracellular Signals That Control Cell Proliferation in Mammalian Balance Epithelia: Key Roles for Phosphatidylinositol-3 Kinase, Mammalian Target of Rapamycin, and S6 Kinases in Preference to Calcium, Protein Kinase C, and Mitogen-Activated Protein Kinase. J. Neurosci. 21: 570-580 [Abstract] [Full Text]  
  • Varlamov, O, Kalinina, E, Che, F., Fricker, L. (2001). Protein phosphatase 2A binds to the cytoplasmic tail of carboxypeptidase D and regulates post-trans-Golgi network trafficking. J. Cell Sci. 114: 311-322 [Abstract]  
  • Page, K., Li, J., Wang, Y., Kartha, S., Pestell, R. G., Hershenson, M. B. (2000). Regulation of Cyclin D1 Expression and DNA Synthesis by Phosphatidylinositol 3-Kinase in Airway Smooth Muscle Cells. Am. J. Respir. Cell Mol. Bio. 23: 436-443 [Abstract] [Full Text]  
  • Law, B. K., Norgaard, P., Moses, H. L. (2000). Farnesyltransferase Inhibitor Induces Rapid Growth Arrest and Blocks p70s6k Activation by Multiple Stimuli. J. Biol. Chem. 275: 10796-10801 [Abstract] [Full Text]  
  • Kainulainen, V., Sundvall, M., Maatta, J. A., Santiestevan, E., Klagsbrun, M., Elenius, K. (2000). A Natural ErbB4 Isoform That Does Not Activate Phosphoinositide 3-Kinase Mediates Proliferation but Not Survival or Chemotaxis. J. Biol. Chem. 275: 8641-8649 [Abstract] [Full Text]  
  • Busse, D., Doughty, R. S., Ramsey, T. T., Russell, W. E., Price, J. O., Flanagan, W. M., Shawver, L. K., Arteaga, C. L. (2000). Reversible G1 Arrest Induced by Inhibition of the Epidermal Growth Factor Receptor Tyrosine Kinase Requires Up-regulation of p27KIP1 Independent of MAPK Activity. J. Biol. Chem. 275: 6987-6995 [Abstract] [Full Text]  
  • Desbois-Mouthon, C., Cadoret, A., Blivet-Van Eggelpoel, M.-J., Bertrand, F., Caron, M., Atfi, A., Cherqui, G., Capeau, J. (2000). Insulin-Mediated Cell Proliferation and Survival Involve Inhibition of c-Jun N-terminal Kinases through a Phosphatidylinositol 3-Kinase- and Mitogen-Activated Protein Kinase Phosphatase-1-Dependent Pathway. Endocrinology 141: 922-931 [Abstract] [Full Text]  
  • Hooshmand-Rad, R, Hajkova, L, Klint, P, Karlsson, R, Vanhaesebroeck, B, Claesson-Welsh, L, Heldin, C. (2000). The PI 3-kinase isoforms p110(alpha) and p110(beta) have differential roles in PDGF- and insulin-mediated signaling. J. Cell Sci. 113: 207-214 [Abstract]  
  • Band, C. J., Mounier, C., Posner, B. I. (1999). Epidermal Growth Factor and Insulin-Induced Deoxyribonucleic Acid Synthesis in Primary Rat Hepatocytes Is Phosphatidylinositol 3-Kinase Dependent and Dissociated from Protooncogene Induction. Endocrinology 140: 5626-5634 [Abstract] [Full Text]  
  • Cass, L. A., Summers, S. A., Prendergast, G. V., Backer, J. M., Birnbaum, M. J., Meinkoth, J. L. (1999). Protein Kinase A-Dependent and -Independent Signaling Pathways Contribute to Cyclic AMP-Stimulated Proliferation. Mol. Cell. Biol. 19: 5882-5891 [Abstract] [Full Text]  
  • Krymskaya, V. P., Penn, R. B., Orsini, M. J., Scott, P. H., Plevin, R. J., Walker, T. R., Eszterhas, A. J., Amrani, Y., Chilvers, E. R., Panettieri, R. A. Jr. (1999). Phosphatidylinositol 3-kinase mediates mitogen-induced human airway smooth muscle cell proliferation. Am. J. Physiol. Lung Cell. Mol. Physiol. 277: L65-L78 [Abstract] [Full Text]  
  • Craddock, B. L., Orchiston, E. A., Hinton, H. J., Welham, M. J. (1999). Dissociation of Apoptosis from Proliferation, Protein Kinase B Activation, and BAD Phosphorylation in Interleukin-3-mediated Phosphoinositide 3-Kinase Signaling. J. Biol. Chem. 274: 10633-10640 [Abstract] [Full Text]  
  • Li, H.-L., Davis, W., Pure, E. (1999). Suboptimal Cross-linking of Antigen Receptor Induces Syk-dependent Activation of p70S6 Kinase through Protein Kinase C and Phosphoinositol 3-Kinase. J. Biol. Chem. 274: 9812-9820 [Abstract] [Full Text]  
  • Shibayama, H., Anzai, N., Braun, S. E., Fukuda, S., Mantel, C., Broxmeyer, H. E. (1999). H-Ras Is Involved in the Inside-out Signaling Pathway of Interleukin-3-Induced Integrin Activation. Blood 93: 1540-1548 [Abstract] [Full Text]  
  • Law, B. K., Norgaard, P., Gnudi, L., Kahn, B. B., Poulson, H. S., Moses, H. L. (1999). Inhibition of DNA Synthesis by a Farnesyltransferase Inhibitor Involves Inhibition of the p70s6k Pathway. J. Biol. Chem. 274: 4743-4748 [Abstract] [Full Text]  
  • Takuwa, N., Fukui, Y., Takuwa, Y. (1999). Cyclin D1 Expression Mediated by Phosphatidylinositol 3-Kinase through mTOR-p70S6K-Independent Signaling in Growth Factor-Stimulated NIH 3T3 Fibroblasts. Mol. Cell. Biol. 19: 1346-1358 [Abstract] [Full Text]  
  • Siddhanta, U., McIlroy, J., Shah, A., Zhang, Y., Backer, J. M. (1998). Distinct Roles for the p110{alpha} and hVPS34 Phosphatidylinositol 3'-Kinases in Vesicular Trafficking, Regulation of the Actin Cytoskeleton, and Mitogenesis. JCB 143: 1647-1659 [Abstract] [Full Text]  
  • Roche, S., Downward, J., Raynal, P., Courtneidge, S. A. (1998). A Function for Phosphatidylinositol 3-Kinase beta  (p85alpha -p110beta ) in Fibroblasts during Mitogenesis: Requirement for Insulin- and Lysophosphatidic Acid-Mediated Signal Transduction. Mol. Cell. Biol. 18: 7119-7129 [Abstract] [Full Text]  
  • Yu, J., Wjasow, C., Backer, J. M. (1998). Regulation of the p85/p110alpha Phosphatidylinositol 3'-Kinase. DISTINCT ROLES FOR THE N-TERMINAL AND C-TERMINAL SH2 DOMAINS. J. Biol. Chem. 273: 30199-30203 [Abstract] [Full Text]  
  • Kennedy, A. P., Sekulic, A., Irvin, B. J., Nilson, A. E., Dilworth, S. M., Abraham, R. T. (1998). Polyomavirus Middle T Antigen as a Probe for T Cell Antigen Receptor-coupled Signaling Pathways. J. Biol. Chem. 273: 11505-11513 [Abstract] [Full Text]  
  • Yu, J., Zhang, Y., McIlroy, J., Rordorf-Nikolic, T., Orr, G. A., Backer, J. M. (1998). Regulation of the p85/p110 Phosphatidylinositol 3'-Kinase: Stabilization and Inhibition of the p110alpha Catalytic Subunit by the p85 Regulatory Subunit. Mol. Cell. Biol. 18: 1379-1387 [Abstract] [Full Text]  
  • Conus, N. M., Hemmings, B. A., Pearson, R. B. (1998). Differential Regulation by Calcium Reveals Distinct Signaling Requirements for the Activation of Akt and p70S6k. J. Biol. Chem. 273: 4776-4782 [Abstract] [Full Text]  
  • Winkler, D. G., Johnson, J. C., Cooper, J. A., Vojtek, A. B. (1997). Identification and Characterization of Mutations in Ha-Ras That Selectively Decrease Binding to cRaf-1. J. Biol. Chem. 272: 24402-24409 [Abstract] [Full Text]  
  • Jascur, T., Gilman, J., Mustelin, T. (1997). Involvement of Phosphatidylinositol 3-Kinase in NFAT Activation in T Cells. J. Biol. Chem. 272: 14483-14488 [Abstract] [Full Text]  
  • Guillemot, L., Levy, A., Zhao, Z. J., Bereziat, G., Rothhut, B. (2000). The Protein-tyrosine Phosphatase SHP-2 Is Required during Angiotensin II-mediated Activation of Cyclin D1 Promoter in CHO-AT1A Cells. J. Biol. Chem. 275: 26349-26358 [Abstract] [Full Text]  
  • Law, B. K., Waltner-Law, M. E., Entingh, A. J., Chytil, A., Aakre, M. E., Norgaard, P., Moses, H. L. (2000). Salicylate-induced Growth Arrest Is Associated with Inhibition of p70s6k and Down-regulation of c-Myc, Cyclin D1, Cyclin A, and Proliferating Cell Nuclear Antigen. J. Biol. Chem. 275: 38261-38267 [Abstract] [Full Text]  
  • Kong, M., Mounier, C., Wu, J., Posner, B. I. (2000). Epidermal Growth Factor-induced Phosphatidylinositol 3-Kinase Activation and DNA Synthesis. IDENTIFICATION OF Grb2-ASSOCIATED BINDER 2 AS THE MAJOR MEDIATOR IN RAT HEPATOCYTES. J. Biol. Chem. 275: 36035-36042 [Abstract] [Full Text]  
  • Hill, K. M., Huang, Y., Yip, S.-C., Yu, J., Segall, J. E., Backer, J. M. (2001). N-terminal Domains of the Class IA Phosphoinositide 3-Kinase Regulatory Subunit Play a Role in Cytoskeletal but Not Mitogenic Signaling. J. Biol. Chem. 276: 16374-16378 [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»