| Mol Cell Biol. 1994 April; 14(4): 2577–2583. | PMCID: PMC358625 |
Direct association of p110 beta phosphatidylinositol 3-kinase with p85 is mediated by an N-terminal fragment of p110 beta. P Hu and J Schlessinger Department of Pharmacology, New York University Medical Center, NY 10016. Abstract Phosphatidylinositol (PI) 3-kinase is a heterodimeric enzyme of 85-kDa (p85) and 110-kDa (p110) subunits implicated in mitogenic signal transduction by virtue of its activation in cells transformed by diverse viral oncoproteins and treated with various growth factors. We have identified a domain in p110 that mediates association with p85 in vitro and in intact cells. A glutathione S-transferase fusion protein containing the N-terminal 171 amino-acids of p110 beta bound to free p85 in cell lysates. This fusion protein also bound directly to p85 immobilized on nitrocellulose filters. An epitope-tagged fragment containing amino acids 31 to 150 of p110 beta associated with p85 upon expression in intact cells. Expression of either an N-terminal fragment of p110 beta or the p85 inter-SH2 domain, which mediates association with p110, reduced the association of endogenous PI 3-kinase activity with the activated platelet-derived growth factor receptor in intact cells. Hence, these defined regions of p85 and p110 mediate the interaction between the two subunits of PI 3-kinase. Full text Full text is available as a scanned copy of the original print version. Get a printable copy (PDF file) of the complete article (1.6M), or click on a page image below to browse page by page. Links to PubMed are also available for Selected References. Images in this article Click on the image to see a larger version. These references are in PubMed. This may not be the complete list of references from this article. - Anderson, D; Koch, CA; Grey, L; Ellis, C; Moran, MF; Pawson, T. Binding of SH2 domains of phospholipase C gamma 1, GAP, and Src to activated growth factor receptors. Science. 1990 Nov 16;250(4983):979–982. [PubMed]
- Auger, KR; Serunian, LA; Soltoff, SP; Libby, P; Cantley, LC. PDGF-dependent tyrosine phosphorylation stimulates production of novel polyphosphoinositides in intact cells. Cell. 1989 Apr 7;57(1):167–175. [PubMed]
- Carpenter, CL; Duckworth, BC; Auger, KR; Cohen, B; Schaffhausen, BS; Cantley, LC. Purification and characterization of phosphoinositide 3-kinase from rat liver. J Biol Chem. 1990 Nov 15;265(32):19704–19711. [PubMed]
- Carter, AN; Downes, CP. Phosphatidylinositol 3-kinase is activated by nerve growth factor and epidermal growth factor in PC12 cells. J Biol Chem. 1992 Jul 25;267(21):14563–14567. [PubMed]
- Chen, C; Okayama, H. High-efficiency transformation of mammalian cells by plasmid DNA. Mol Cell Biol. 1987 Aug;7(8):2745–2752. [PubMed]
- Corey, S; Eguinoa, A; Puyana-Theall, K; Bolen, JB; Cantley, L; Mollinedo, F; Jackson, TR; Hawkins, PT; Stephens, LR. Granulocyte macrophage-colony stimulating factor stimulates both association and activation of phosphoinositide 3OH-kinase and src-related tyrosine kinase(s) in human myeloid derived cells. EMBO J. 1993 Jul;12(7):2681–2690. [PubMed]
- Coughlin, SR; Escobedo, JA; Williams, LT. Role of phosphatidylinositol kinase in PDGF receptor signal transduction. Science. 1989 Mar 3;243(4895):1191–1194. [PubMed]
- Courtneidge, SA; Heber, A. An 81 kd protein complexed with middle T antigen and pp60c-src: a possible phosphatidylinositol kinase. Cell. 1987 Sep 25;50(7):1031–1037. [PubMed]
- Escobedo, JA; Navankasattusas, S; Kavanaugh, WM; Milfay, D; Fried, VA; Williams, LT. cDNA cloning of a novel 85 kd protein that has SH2 domains and regulates binding of PI3-kinase to the PDGF beta-receptor. Cell. 1991 Apr 5;65(1):75–82. [PubMed]
- Fantl, WJ; Escobedo, JA; Martin, GA; Turck, CW; del Rosario, M; McCormick, F; Williams, LT. Distinct phosphotyrosines on a growth factor receptor bind to specific molecules that mediate different signaling pathways. Cell. 1992 May 1;69(3):413–423. [PubMed]
- Field, J; Nikawa, J; Broek, D; MacDonald, B; Rodgers, L; Wilson, IA; Lerner, RA; Wigler, M. Purification of a RAS-responsive adenylyl cyclase complex from Saccharomyces cerevisiae by use of an epitope addition method. Mol Cell Biol. 1988 May;8(5):2159–2165. [PubMed]
- Fukui, Y; Hanafusa, H. Phosphatidylinositol kinase activity associates with viral p60src protein. Mol Cell Biol. 1989 Apr;9(4):1651–1658. [PubMed]
- Fukui, Y; Kornbluth, S; Jong, SM; Wang, LH; Hanafusa, H. Phosphatidylinositol kinase type I activity associates with various oncogene products. Oncogene Res. 1989;4(4):283–292. [PubMed]
- Fukui, Y; Saltiel, AR; Hanafusa, H. Phosphatidylinositol-3 kinase is activated in v-src, v-yes, and v-fps transformed chicken embryo fibroblasts. Oncogene. 1991 Mar;6(3):407–411. [PubMed]
- Graziani, A; Gramaglia, D; Cantley, LC; Comoglio, PM. The tyrosine-phosphorylated hepatocyte growth factor/scatter factor receptor associates with phosphatidylinositol 3-kinase. J Biol Chem. 1991 Nov 25;266(33):22087–22090. [PubMed]
- Hiles, ID; Otsu, M; Volinia, S; Fry, MJ; Gout, I; Dhand, R; Panayotou, G; Ruiz-Larrea, F; Thompson, A; Totty, NF, et al. Phosphatidylinositol 3-kinase: structure and expression of the 110 kd catalytic subunit. Cell. 1992 Aug 7;70(3):419–429. [PubMed]
- Hu, P; Margolis, B; Skolnik, EY; Lammers, R; Ullrich, A; Schlessinger, J. Interaction of phosphatidylinositol 3-kinase-associated p85 with epidermal growth factor and platelet-derived growth factor receptors. Mol Cell Biol. 1992 Mar;12(3):981–990. [PubMed]
- Hu, P; Mondino, A; Skolnik, EY; Schlessinger, J. Cloning of a novel, ubiquitously expressed human phosphatidylinositol 3-kinase and identification of its binding site on p85. Mol Cell Biol. 1993 Dec;13(12):7677–7688. [PubMed]
- Jackson, TR; Stephens, LR; Hawkins, PT. Receptor specificity of growth factor-stimulated synthesis of 3-phosphorylated inositol lipids in Swiss 3T3 cells. J Biol Chem. 1992 Aug 15;267(23):16627–16636. [PubMed]
- Kaplan, DR; Whitman, M; Schaffhausen, B; Pallas, DC; White, M; Cantley, L; Roberts, TM. Common elements in growth factor stimulation and oncogenic transformation: 85 kd phosphoprotein and phosphatidylinositol kinase activity. Cell. 1987 Sep 25;50(7):1021–1029. [PubMed]
- Kaplan, DR; Whitman, M; Schaffhausen, B; Raptis, L; Garcea, RL; Pallas, D; Roberts, TM; Cantley, L. Phosphatidylinositol metabolism and polyoma-mediated transformation. Proc Natl Acad Sci U S A. 1986 Jun;83(11):3624–3628. [PubMed]
- Kashishian, A; Kazlauskas, A; Cooper, JA. Phosphorylation sites in the PDGF receptor with different specificities for binding GAP and PI3 kinase in vivo. EMBO J. 1992 Apr;11(4):1373–1382. [PubMed]
- Klippel, A; Escobedo, JA; Fantl, WJ; Williams, LT. The C-terminal SH2 domain of p85 accounts for the high affinity and specificity of the binding of phosphatidylinositol 3-kinase to phosphorylated platelet-derived growth factor beta receptor. Mol Cell Biol. 1992 Apr;12(4):1451–1459. [PubMed]
- Klippel, A; Escobedo, JA; Hu, Q; Williams, LT. A region of the 85-kilodalton (kDa) subunit of phosphatidylinositol 3-kinase binds the 110-kDa catalytic subunit in vivo. Mol Cell Biol. 1993 Sep;13(9):5560–5566. [PubMed]
- Kozak, M. The scanning model for translation: an update. J Cell Biol. 1989 Feb;108(2):229–241. [PubMed]
- Lev, S; Givol, D; Yarden, Y. A specific combination of substrates is involved in signal transduction by the kit-encoded receptor. EMBO J. 1991 Mar;10(3):647–654. [PubMed]
- Lev, S; Givol, D; Yarden, Y. Interkinase domain of kit contains the binding site for phosphatidylinositol 3' kinase. Proc Natl Acad Sci U S A. 1992 Jan 15;89(2):678–682. [PubMed]
- Ling, LE; Druker, BJ; Cantley, LC; Roberts, TM. Transformation-defective mutants of polyomavirus middle T antigen associate with phosphatidylinositol 3-kinase (PI 3-kinase) but are unable to maintain wild-type levels of PI 3-kinase products in intact cells. J Virol. 1992 Mar;66(3):1702–1708. [PubMed]
- Lips, DL; Majerus, PW; Gorga, FR; Young, AT; Benjamin, TL. Phosphatidylinositol 3-phosphate is present in normal and transformed fibroblasts and is resistant to hydrolysis by bovine brain phospholipase C II. J Biol Chem. 1989 May 25;264(15):8759–8763. [PubMed]
- McGlade, CJ; Ellis, C; Reedijk, M; Anderson, D; Mbamalu, G; Reith, AD; Panayotou, G; End, P; Bernstein, A; Kazlauskas, A, et al. SH2 domains of the p85 alpha subunit of phosphatidylinositol 3-kinase regulate binding to growth factor receptors. Mol Cell Biol. 1992 Mar;12(3):991–997. [PubMed]
- Moran, MF; Koch, CA; Anderson, D; Ellis, C; England, L; Martin, GS; Pawson, T. Src homology region 2 domains direct protein-protein interactions in signal transduction. Proc Natl Acad Sci U S A. 1990 Nov;87(21):8622–8626. [PubMed]
- Myers, MG, Jr; Backer, JM; Sun, XJ; Shoelson, S; Hu, P; Schlessinger, J; Yoakim, M; Schaffhausen, B; White, MF. IRS-1 activates phosphatidylinositol 3'-kinase by associating with src homology 2 domains of p85. Proc Natl Acad Sci U S A. 1992 Nov 1;89(21):10350–10354. [PubMed]
- Nishimura, R; Li, W; Kashishian, A; Mondino, A; Zhou, M; Cooper, J; Schlessinger, J. Two signaling molecules share a phosphotyrosine-containing binding site in the platelet-derived growth factor receptor. Mol Cell Biol. 1993 Nov;13(11):6889–6896. [PubMed]
- Otsu, M; Hiles, I; Gout, I; Fry, MJ; Ruiz-Larrea, F; Panayotou, G; Thompson, A; Dhand, R; Hsuan, J; Totty, N, et al. Characterization of two 85 kd proteins that associate with receptor tyrosine kinases, middle-T/pp60c-src complexes, and PI3-kinase. Cell. 1991 Apr 5;65(1):91–104. [PubMed]
- Peles, E; Lamprecht, R; Ben-Levy, R; Tzahar, E; Yarden, Y. Regulated coupling of the Neu receptor to phosphatidylinositol 3'-kinase and its release by oncogenic activation. J Biol Chem. 1992 Jun 15;267(17):12266–12274. [PubMed]
- Remillard, B; Petrillo, R; Maslinski, W; Tsudo, M; Strom, TB; Cantley, L; Varticovski, L. Interleukin-2 receptor regulates activation of phosphatidylinositol 3-kinase. J Biol Chem. 1991 Aug 5;266(22):14167–14170. [PubMed]
- Ren, R; Mayer, BJ; Cicchetti, P; Baltimore, D. Identification of a ten-amino acid proline-rich SH3 binding site. Science. 1993 Feb 19;259(5098):1157–1161. [PubMed]
- Rottapel, R; Reedijk, M; Williams, DE; Lyman, SD; Anderson, DM; Pawson, T; Bernstein, A. The Steel/W transduction pathway: kit autophosphorylation and its association with a unique subset of cytoplasmic signaling proteins is induced by the Steel factor. Mol Cell Biol. 1991 Jun;11(6):3043–3051. [PubMed]
- Ruderman, NB; Kapeller, R; White, MF; Cantley, LC. Activation of phosphatidylinositol 3-kinase by insulin. Proc Natl Acad Sci U S A. 1990 Feb;87(4):1411–1415. [PubMed]
- Serunian, LA; Auger, KR; Roberts, TM; Cantley, LC. Production of novel polyphosphoinositides in vivo is linked to cell transformation by polyomavirus middle T antigen. J Virol. 1990 Oct;64(10):4718–4725. [PubMed]
- Serunian, LA; Haber, MT; Fukui, T; Kim, JW; Rhee, SG; Lowenstein, JM; Cantley, LC. Polyphosphoinositides produced by phosphatidylinositol 3-kinase are poor substrates for phospholipases C from rat liver and bovine brain. J Biol Chem. 1989 Oct 25;264(30):17809–17815. [PubMed]
- Shibasaki, F; Homma, Y; Takenawa, T. Two types of phosphatidylinositol 3-kinase from bovine thymus. Monomer and heterodimer form. J Biol Chem. 1991 May 5;266(13):8108–8114. [PubMed]
- Skolnik, EY; Margolis, B; Mohammadi, M; Lowenstein, E; Fischer, R; Drepps, A; Ullrich, A; Schlessinger, J. Cloning of PI3 kinase-associated p85 utilizing a novel method for expression/cloning of target proteins for receptor tyrosine kinases. Cell. 1991 Apr 5;65(1):83–90. [PubMed]
- Smith, DB; Johnson, KS. Single-step purification of polypeptides expressed in Escherichia coli as fusions with glutathione S-transferase. Gene. 1988 Jul 15;67(1):31–40. [PubMed]
- Soltoff, SP; Rabin, SL; Cantley, LC; Kaplan, DR. Nerve growth factor promotes the activation of phosphatidylinositol 3-kinase and its association with the trk tyrosine kinase. J Biol Chem. 1992 Aug 25;267(24):17472–17477. [PubMed]
- Talmage, DA; Freund, R; Young, AT; Dahl, J; Dawe, CJ; Benjamin, TL. Phosphorylation of middle T by pp60c-src: a switch for binding of phosphatidylinositol 3-kinase and optimal tumorigenesis. Cell. 1989 Oct 6;59(1):55–65. [PubMed]
- Tyers, M; Tokiwa, G; Nash, R; Futcher, B. The Cln3-Cdc28 kinase complex of S. cerevisiae is regulated by proteolysis and phosphorylation. EMBO J. 1992 May;11(5):1773–1784. [PubMed]
- Ulug, ET; Hawkins, PT; Hanley, MR; Courtneidge, SA. Phosphatidylinositol metabolism in cells transformed by polyomavirus middle T antigen. J Virol. 1990 Aug;64(8):3895–3904. [PubMed]
- Varticovski, L; Daley, GQ; Jackson, P; Baltimore, D; Cantley, LC. Activation of phosphatidylinositol 3-kinase in cells expressing abl oncogene variants. Mol Cell Biol. 1991 Feb;11(2):1107–1113. [PubMed]
- Varticovski, L; Druker, B; Morrison, D; Cantley, L; Roberts, T. The colony stimulating factor-1 receptor associates with and activates phosphatidylinositol-3 kinase. Nature. 1989 Dec 7;342(6250):699–702. [PubMed]
- Whitman, M; Downes, CP; Keeler, M; Keller, T; Cantley, L. Type I phosphatidylinositol kinase makes a novel inositol phospholipid, phosphatidylinositol-3-phosphate. Nature. 1988 Apr 14;332(6165):644–646. [PubMed]
- Whitman, M; Kaplan, DR; Schaffhausen, B; Cantley, L; Roberts, TM. Association of phosphatidylinositol kinase activity with polyoma middle-T competent for transformation. Nature. 315(6016):239–242. [PubMed]
- Zippel, R; Sturani, E; Toschi, L; Naldini, L; Alberghina, L; Comoglio, PM. In vivo phosphorylation and dephosphorylation of the platelet-derived growth factor receptor studied by immunoblot analysis with phosphotyrosine antibodies. Biochim Biophys Acta. 1986 Mar 19;881(1):54–61. [PubMed]
|
This article has been 
