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

 Previous Article  |  Next Article 

Molecular and Cellular Biology, November 2000, p. 8035-8046, Vol. 20, No. 21
0270-7306/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.

Positive and Negative Regulation of Phosphoinositide 3-Kinase-Dependent Signaling Pathways by Three Different Gene Products of the p85 Regulatory Subunit

Kohjiro Ueki, Petra Algenstaedt, Franck Mauvais-Jarvis, and C. Ronald Kahn^*

Research Division, Joslin Diabetes Center, Harvard Medical School, Boston, Massachusetts 02215

Received 23 March 2000/Returned for modification 8 May 2000/Accepted 9 August 2000

Phosphoinositide (PI) 3-kinase is a key mediator of insulin-dependent metabolic actions, including stimulation of glucose transport and glycogen synthesis. The gene for the p85 regulatory subunit yields three splicing variants, p85, AS53/p55, and p50. All three have (i) a C-terminal structure consisting of two Src homology 2 domains flanking the p110 catalytic subunit-binding domain and (ii) a unique N-terminal region of 304, 34, and 6 amino acids, respectively. To determine if these regulatory subunits differ in their effects on enzyme activity and signal transduction from insulin receptor substrate (IRS) proteins under physiological conditions, we expressed each regulatory subunit in fully differentiated L6 myotubes using adenovirus-mediated gene transfer with or without coexpression of the p110 catalytic subunit. PI 3-kinase activity associated with p50 was greater than that associated with p85 or AS53. Increasing the level of p85 or AS53, but not p50, inhibited both phosphotyrosine-associated and p110-associated PI 3-kinase activities. Expression of a p85 mutant lacking the p110-binding site (p85) also inhibited phosphotyrosine-associated PI 3-kinase activity but not p110-associated activity. Insulin stimulation of two kinases downstream from PI-3 kinase, Akt and p70 S6 kinase (p70^S6K), was decreased in cells expressing p85 or AS53 but not in cells expressing p50. Similar inhibition of PI 3-kinase, Akt, and p70^S6K was observed, even when p110 was coexpressed with p85 or AS53. Expression of p110 alone dramatically increased glucose transport but decreased glycogen synthase activity. This effect was reduced when p110 was coexpressed with any of the three regulatory subunits. Thus, the three different isoforms of regulatory subunit can relay the signal from IRS proteins to the p110 catalytic subunit with different efficiencies. They also negatively modulate the PI 3-kinase catalytic activity but to different extents, dependent on the unique N-terminal structure of each isoform. These data also suggest the existence of a mechanism by which regulatory subunits modulate the PI 3-kinase-mediated signals, independent of the kinase activity, possibly through subcellular localization of the catalytic subunit or interaction with additional signaling molecules.

---

^* Corresponding author. Mailing address: Joslin Diabetes Center, One Joslin Place, Boston, MA 02215. Phone: (617) 732-2635. Fax: (617) 732-2593. E-mail: c.ronald.kahn{at}joslin.harvard.edu.

---

Molecular and Cellular Biology, November 2000, p. 8035-8046, Vol. 20, No. 21
0270-7306/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.

This article has been cited by other articles:

• Xie, D., Gore, C., Zhou, J., Pong, R.-C., Zhang, H., Yu, L., Vessella, R. L., Min, W., Hsieh, J.-T. (2009). DAB2IP coordinates both PI3K-Akt and ASK1 pathways for cell survival and apoptosis. Proc. Natl. Acad. Sci. USA 106: 19878-19883 [Abstract] [Full Text]  
• Assmann, A., Ueki, K., Winnay, J. N., Kadowaki, T., Kulkarni, R. N. (2009). Glucose Effects on Beta-Cell Growth and Survival Require Activation of Insulin Receptors and Insulin Receptor Substrate 2. Mol. Cell. Biol. 29: 3219-3228 [Abstract] [Full Text]  
• Kim, J. J., Choi, Y. M., Hong, M. A., Hwang, S. S., Yoon, S. H., Chae, S. J., Jee, B. C., Ku, S. Y., Kim, J. G., Moon, S. Y. (2009). Phosphatidylinositol 3-kinase p85{alpha} regulatory subunit gene Met326Ile polymorphism in women with polycystic ovary syndrome. Hum Reprod 24: 1184-1190 [Abstract] [Full Text]  
• Aoki, K., Matsui, J., Kubota, N., Nakajima, H., Iwamoto, K., Takamoto, I., Tsuji, Y., Ohno, A., Mori, S., Tokuyama, K., Murakami, K., Asano, T., Aizawa, S., Tobe, K., Kadowaki, T., Terauchi, Y. (2009). Role of the liver in glucose homeostasis in PI 3-kinase p85{alpha}-deficient mice. Am. J. Physiol. Endocrinol. Metab. 296: E842-E853 [Abstract] [Full Text]  
• Adochio, R., Leitner, J. W., Hedlund, R., Draznin, B. (2009). Rescuing 3T3-L1 Adipocytes from Insulin Resistance Induced by Stimulation of Akt-Mammalian Target of Rapamycin/p70 S6 Kinase (S6K1) Pathway and Serine Phosphorylation of Insulin Receptor Substrate-1: Effect of Reduced Expression of p85{alpha} Subunit of Phosphatidylinositol 3-Kinase and S6K1 Kinase. Endocrinology 150: 1165-1173 [Abstract] [Full Text]  
• Takeda, J.-i., Suzuki, Y., Sakate, R., Sato, Y., Seki, M., Irie, T., Takeuchi, N., Ueda, T., Nakao, M., Sugano, S., Gojobori, T., Imanishi, T. (2008). Low conservation and species-specific evolution of alternative splicing in humans and mice: comparative genomics analysis using well-annotated full-length cDNAs. Nucleic Acids Res 36: 6386-6395 [Abstract] [Full Text]  
• Fos, C., Salles, A., Lang, V., Carrette, F., Audebert, S., Pastor, S., Ghiotto, M., Olive, D., Bismuth, G., Nunes, J. A. (2008). ICOS Ligation Recruits the p50{alpha} PI3K Regulatory Subunit to the Immunological Synapse. J. Immunol. 181: 1969-1977 [Abstract] [Full Text]  
• Friedman, J. E., Kirwan, J. P., Jing, M., Presley, L., Catalano, P. M. (2008). Increased Skeletal Muscle Tumor Necrosis Factor-{alpha} and Impaired Insulin Signaling Persist in Obese Women With Gestational Diabetes Mellitus 1 Year Postpartum. Diabetes 57: 606-613 [Abstract] [Full Text]  
• del Rincon, J.-P., Iida, K., Gaylinn, B. D., McCurdy, C. E., Leitner, J. W., Barbour, L. A., Kopchick, J. J., Friedman, J. E., Draznin, B., Thorner, M. O. (2007). Growth Hormone Regulation of p85{alpha} Expression and Phosphoinositide 3-Kinase Activity in Adipose Tissue: Mechanism for Growth Hormone-Mediated Insulin Resistance. Diabetes 56: 1638-1646 [Abstract] [Full Text]  
• Geering, B., Cutillas, P. R., Nock, G., Gharbi, S. I., Vanhaesebroeck, B. (2007). Class IA phosphoinositide 3-kinases are obligate p85-p110 heterodimers. Proc. Natl. Acad. Sci. USA 104: 7809-7814 [Abstract] [Full Text]  
• Taniguchi, C. M., Aleman, J. O., Ueki, K., Luo, J., Asano, T., Kaneto, H., Stephanopoulos, G., Cantley, L. C., Kahn, C. R. (2007). The p85{alpha} Regulatory Subunit of Phosphoinositide 3-Kinase Potentiates c-Jun N-Terminal Kinase-Mediated Insulin Resistance. Mol. Cell. Biol. 27: 2830-2840 [Abstract] [Full Text]  
• Spender, L. C., Lucchesi, W., Bodelon, G., Bilancio, A., Karstegl, C. E., Asano, T., Dittrich-Breiholz, O., Kracht, M., Vanhaesebroeck, B., Farrell, P. J. (2006). Cell target genes of Epstein-Barr virus transcription factor EBNA-2: induction of the p55{alpha} regulatory subunit of PI3-kinase and its role in survival of EREB2.5 cells.. J. Gen. Virol. 87: 2859-2867 [Abstract] [Full Text]  
• Draznin, B. (2006). Molecular Mechanisms of Insulin Resistance: Serine Phosphorylation of Insulin Receptor Substrate-1 and Increased Expression of p85{alpha}: The Two Sides of a Coin.. Diabetes 55: 2392-2397 [Abstract] [Full Text]  
• Yang, M.-H., Chiang, W.-C., Chou, T.-Y., Chang, S.-Y., Chen, P.-M., Teng, S.-C., Wu, K.-J. (2006). Increased NBS1 Expression Is a Marker of Aggressive Head and Neck Cancer and Overexpression of NBS1 Contributes to Transformation. Clin. Cancer Res. 12: 507-515 [Abstract] [Full Text]  
• Barbour, L. A., Mizanoor Rahman, S., Gurevich, I., Leitner, J. W., Fischer, S. J., Roper, M. D., Knotts, T. A., Vo, Y., McCurdy, C. E., Yakar, S., LeRoith, D., Kahn, C. R., Cantley, L. C., Friedman, J. E., Draznin, B. (2005). Increased P85{alpha} Is a Potent Negative Regulator of Skeletal Muscle Insulin Signaling and Induces in Vivo Insulin Resistance Associated with Growth Hormone Excess. J. Biol. Chem. 280: 37489-37494 [Abstract] [Full Text]  
• Chen, Y.-C., Su, Y.-N., Chou, P.-C., Chiang, W.-C., Chang, M.-C., Wang, L.-S., Teng, S.-C., Wu, K.-J. (2005). Overexpression of NBS1 Contributes to Transformation through the Activation of Phosphatidylinositol 3-Kinase/Akt. J. Biol. Chem. 280: 32505-32511 [Abstract] [Full Text]  
• Luo, J., Field, S. J., Lee, J. Y., Engelman, J. A., Cantley, L. C. (2005). The p85 regulatory subunit of phosphoinositide 3-kinase down-regulates IRS-1 signaling via the formation of a sequestration complex. JCB 170: 455-464 [Abstract] [Full Text]  
• Bandyopadhyay, G. K., Yu, J. G., Ofrecio, J., Olefsky, J. M. (2005). Increased p85/55/50 Expression and Decreased Phosphotidylinositol 3-Kinase Activity in Insulin-Resistant Human Skeletal Muscle. Diabetes 54: 2351-2359 [Abstract] [Full Text]  
• Fabre, S., Lang, V., Harriague, J., Jobart, A., Unterman, T. G., Trautmann, A., Bismuth, G. (2005). Stable Activation of Phosphatidylinositol 3-Kinase in the T Cell Immunological Synapse Stimulates Akt Signaling to FoxO1 Nuclear Exclusion and Cell Growth Control. J. Immunol. 174: 4161-4171 [Abstract] [Full Text]  
• Brachmann, S. M., Ueki, K., Engelman, J. A., Kahn, R. C., Cantley, L. C. (2005). Phosphoinositide 3-Kinase Catalytic Subunit Deletion and Regulatory Subunit Deletion Have Opposite Effects on Insulin Sensitivity in Mice. Mol. Cell. Biol. 25: 1596-1607 [Abstract] [Full Text]  
• Hay, N., Sonenberg, N. (2004). Upstream and downstream of mTOR. Genes Dev. 18: 1926-1945 [Abstract] [Full Text]  
• Barbour, L. A., Shao, J., Qiao, L., Leitner, W., Anderson, M., Friedman, J. E., Draznin, B. (2004). Human Placental Growth Hormone Increases Expression of the P85 Regulatory Unit of Phosphatidylinositol 3-Kinase and Triggers Severe Insulin Resistance in Skeletal Muscle. Endocrinology 145: 1144-1150 [Abstract] [Full Text]  
• Chen, D., Mauvais-Jarvis, F., Bluher, M., Fisher, S. J., Jozsi, A., Goodyear, L. J., Ueki, K., Kahn, C. R. (2004). p50{alpha}/p55{alpha} Phosphoinositide 3-Kinase Knockout Mice Exhibit Enhanced Insulin Sensitivity. Mol. Cell. Biol. 24: 320-329 [Abstract] [Full Text]  
• Harris, T. E., Lawrence, J. C. Jr. (2003). TOR Signaling. Sci Signal 2003: re15-re15 [Abstract] [Full Text]  
• Ueki, K., Fruman, D. A., Yballe, C. M., Fasshauer, M., Klein, J., Asano, T., Cantley, L. C., Kahn, C. R. (2003). Positive and Negative Roles of p85{alpha} and p85{beta} Regulatory Subunits of Phosphoinositide 3-Kinase in Insulin Signaling. J. Biol. Chem. 278: 48453-48466 [Abstract] [Full Text]  
• Lee, H.-Y., Srinivas, H., Xia, D., Lu, Y., Superty, R., LaPushin, R., Gomez-Manzano, C., Gal, A. M., Walsh, G. L., Force, T., Ueki, K., Mills, G. B., Kurie, J. M. (2003). Evidence That Phosphatidylinositol 3-Kinase- and Mitogen-activated Protein Kinase Kinase-4/c-Jun NH2-terminal Kinase-dependent Pathways Cooperate to Maintain Lung Cancer Cell Survival. J. Biol. Chem. 278: 23630-23638 [Abstract] [Full Text]  
• Hallmann, D., Trumper, K., Trusheim, H., Ueki, K., Kahn, C. R., Cantley, L. C., Fruman, D. A., Horsch, D. (2003). Altered Signaling and Cell Cycle Regulation in Embryonal Stem Cells with a Disruption of the Gene for Phosphoinositide 3-Kinase Regulatory Subunit p85alpha. J. Biol. Chem. 278: 5099-5108 [Abstract] [Full Text]  
• Ueki, K., Fruman, D. A., Brachmann, S. M., Tseng, Y.-H., Cantley, L. C., Kahn, C. R. (2002). Molecular Balance between the Regulatory and Catalytic Subunits of Phosphoinositide 3-Kinase Regulates Cell Signaling and Survival. Mol. Cell. Biol. 22: 965-977 [Abstract] [Full Text]  
• Pagliassotti, M. J., Kang, J., Thresher, J. S., Sung, C. K., Bizeau, M. E. (2002). Elevated basal PI 3-kinase activity and reduced insulin signaling in sucrose-induced hepatic insulin resistance. Am. J. Physiol. Endocrinol. Metab. 282: E170-E176 [Abstract] [Full Text]  
• Ueki, K., Yballe, C. M., Brachmann, S. M., Vicent, D., Watt, J. M., Kahn, C. R., Cantley, L. C. (2002). Increased insulin sensitivity in mice lacking p85beta subunit of phosphoinositide 3-kinase. Proc. Natl. Acad. Sci. USA 99: 419-424 [Abstract] [Full Text]