The p110${\gamma}$ PI-3 Kinase is Required for the Mechanism by Which the EphA8-induced Neurites are Modulated by Ephrin-A5 Engagement

  • Park, Soo-Chul (Department of Life Science, College of Sciences, Sookmyung Womens University)
  • Published : 2004.03.01

Abstract

This study provides evidence that expression of EphA8 receptor in NG108-15 cells results in a substantial increase in the number of neurite-bearing cells. However, the EphA8-induced neurite outgrowth does not require either ephrin-A5 stimulation or ectopic expression of $p110{\gamma}$ PI-3 kinase. In contrast, co-expression of a lipid kinase-inactive $p110{\gamma}$ mutant together with EphA8 causes neurite retraction in the presence of ephrin-A5 stimulation. This effect was not observed in the absence of ephrin-A5 stimulation. Significantly, the tyrosine kinase activity of EphA8 is not important for either of these processes. Taken together, our results strongly suggest that $p110{\gamma}$ PI-3 kinase is critically involved in the regulatory process by which ephrin-A5 exerts effects on the EphA8-induced neurite outgrowth.

Keywords

References

  1. Cantley LC (2002) The phosphoinositide 3-kinase pathway. Science 296: 1655-1657 https://doi.org/10.1126/science.296.5573.1655
  2. Davy A, Gale NW, Murray EW, Klinghoffer RA, Soriano P, Feuerstein C, and Robbins SM (1999) Compartmentalized signaling by GPI-anchored ephrin-A5 requires the Fyn tyrosine kinase to regulate cellular adhesion. Genes Dev 13: 3125-3135 https://doi.org/10.1101/gad.13.23.3125
  3. Dodelet VC, Pazzagli C, Zisch AH, Hauser CA, and Pasquale EB (1999) A novel signaling intermediate, SHEP1, directly couples Eph receptors to R-Ras and Rap1A. J Biol Chem 274: 31941-31946 https://doi.org/10.1074/jbc.274.45.31941
  4. Elowe S, Holland SJ, Kulkarni S, and Pawson T (2001) Downregulation of the Rasmitogen-activated protein kinase pathway by the EphB2 receptor tyrosine kinase is required for ephrin-induced neurite retraction. Mol Cell Biol 21 : 7429-7441 https://doi.org/10.1128/MCB.21.21.7429-7441.2001
  5. Gu C and Park S (2001) The EphA8 receptor regulates integrin activity through p110gamma phosphatidylinositol-3 kinase in a tyrosine kinase activity-independent manner. Mol Cell Biol 21: 4579-4597 https://doi.org/10.1128/MCB.21.14.4579-4597.2001
  6. Gu C and Park S (2003) The p110 gamma PI-3 kinase is required for EphA8-stimulated cell migration. FEBS Lett 540: 65-70 https://doi.org/10.1016/S0014-5793(03)00223-0
  7. Holly SP, Larson MK, and Parise LV (2000) Multiple roles of integrins in cell motility. Exp Cell Res 261: 69-74 https://doi.org/10.1006/excr.2000.5040
  8. Huai J and Drescher U (2001) An ephrin-A-dependent signaling pathway controls integrin function and is linked to the tyrosine phosphorylation of a 120-kDa protein. J Biol Chem 276: 6689-6694 https://doi.org/10.1074/jbc.M008127200
  9. Huynh-Do U, Stein E, Lane AA, Liu H, Cerretti DP, and Daniel TO (1999) Surface densities of ephrin-B1 determine EphB1-coupled activation of cell attachment through alphavbeta3 and alpha5beta1 integrins. EMBO J 18: 2165-2173 https://doi.org/10.1093/emboj/18.8.2165
  10. Knoll B, Zarbalis K, Wurst W, and Drescher U (2001) A role for the EphA family in the topographic targeting of vomeronasal axons. Development 128: 895-906
  11. Ma AD, Metjian A, Bagrodia S, Taylor S. and Abrams CS (1998) Cytoskeletal reorganization by G protein-coupled receptors is dependent on phosphoinositide 3-kinase gamma, a Rac guanosine exchange factor, and Rac. Mol Cell Biol 18: 4744-4751 https://doi.org/10.1128/MCB.18.8.4744
  12. Mellitzer G, Xu Q, and Wilkinson DG (2000) Control of cell behaviour by signalling through Eph receptors and ephrins. Curr Opin Neurobiol 10: 400-408 https://doi.org/10.1016/S0959-4388(00)00095-7
  13. Menzel P, Valencia F, Godement P, Dodelet VC, and Pasquale EB (2001) Ephrin-A6, a new ligand for EphA receptors in the developing visual system. Dev Biol 230: 74-88 https://doi.org/10.1006/dbio.2000.0109
  14. Miao H, Burnett E, Kinch M, Simon E, and Wang B (2000) Activation of EphA2 kinase suppresses integrin function and causes focal-adhesion-kinase dephosphorylation. Nat Cell Biol 2: 62-69 https://doi.org/10.1038/35000008
  15. Nelson P, Christian C, and Nirenberg M (1976) Synapse formation between clonal neuroblastoma X glioma hybrid cells and striated muscle cells. Proc Natl Acad Sci USA 73: 123-127 https://doi.org/10.1073/pnas.73.1.123
  16. Nielsen M, Svejgaard A, Skov S, Dobson P, Bendtzen K, Geisler C, and Odum N (1996) IL-2 induces beta2-integrin adhesion via a wortmannin/LY294002-sensitive, rapamycin-resistant pathway. Phosphorylation of a 125-kilodalton protein correlates with induction of adhesion, but not mitogenesis. J Immunol 157: 5350-5358
  17. Park S and Sanchez MP (1997) The Eek receptor, a member of the Eph family of tyrosine protein kinases, can be activated by three different Eph family ligands. Oncogene 14: 533-542 https://doi.org/10.1038/sj.onc.1200857
  18. Sasaki T, Irie-Sasaki J, Jones RG, Oliveira-dos-Santos AJ, Stanford WL, Bolon B, Wakeham A, Itie A, Bouchard D, Kozieradzki I, Joza N, Mak TW, Ohashi PS, Suzuki A, and Penninger JM (2000) Function of PI3Kgamma in thymocyte development, T cell activation, and neutrophil migration. Science 287: 1040-1046 https://doi.org/10.1126/science.287.5455.1040
  19. Shamah SM, Lin MZ, Goldberg JL, Estrach S, Sahin M, Hu L, Bazalakova M, Neve RL, Corfas G, Debant A, and Greenberg ME (2001) EphA receptors regulate growth cone dynamics through the novel guanine nucleotide exchange factor ephexin. Cell 105: 233-244 https://doi.org/10.1016/S0092-8674(01)00314-2
  20. Shimizu Y, Mobley JL, Finkelstein LD, and Chan AS (1995) A role for phosphatidylinositol 3-kinase in the regulation of beta 1 integrin activity by the CD2 antigen. J Cell Biol 131: 1867-1880 https://doi.org/10.1083/jcb.131.6.1867
  21. Smith A, Robinson V, Patel K, and Wilkinson DG (1997) The EphA4 and EphB1 receptor tyrosine kinases and ephrin-B2 ligand regulate targeted migration of branchial neural crest cells. Curr Biol 7: 561-570 https://doi.org/10.1016/S0960-9822(06)00255-7
  22. Tang X and Downes CP (1997) Purification and characterization of Gbetagammaresponsive phosphoinositide 3-kinases from pig platelet cytosol. J Biol Chem 272: 14193-14299 https://doi.org/10.1074/jbc.272.22.14193
  23. Wahl S, Barth H, Ciossek T, Aktories K, and Mueller BK (2000) Ephrin-A5 induces collapse of growth cones by activating Rho and Rho kinase. J Cell Biol 149: 263-270 https://doi.org/10.1083/jcb.149.2.263
  24. Zell T, Hunt SW, Mobley JL, Finkelstein LD, and Shimizu Y (1996) CD28-mediated upregulation of beta 1-integrin adhesion involves phosphatidylinositol 3-kinase. J Immunol 156: 883-886
  25. Zou JX, Wang B, Kalo MS, Zisch AH, Pasquale EB, and Ruoslahti E (1999) An Eph receptor regulates integrin activity through R-Ras. Proc Natl Acad Sci USA 96: 13813-13818 https://doi.org/10.1073/pnas.96.24.13813