Skip to main content
NCBI home page
Proceedings of the National Academy of Sciences of the United States of America logoLink to Proceedings of the National Academy of Sciences of the United States of America
. 1986 Apr;83(8):2353–2357. doi: 10.1073/pnas.83.8.2353

Protein kinase C as a component of a nerve growth factor-sensitive phosphorylation system in PC12 cells.

T Hama, K P Huang, G Guroff
PMCID: PMC323295  PMID: 3458200

Abstract

The treatment of PC12 cells with either nerve growth factor or phorbol 12-myristate 13-acetate caused a decrease in the phosphorylation of a soluble 100-kDa protein (Nsp100). After treatment with nerve growth factor, the activity of Ca2+/phospholipid-dependent protein kinase (protein kinase C) in the cytosol was increased. When the cytosol from untreated PC12 cells was preincubated with purified protein kinase C and its cofactors, the phosphorylation of Nsp100 was decreased. The preincubation of cytosol from nerve growth factor-treated PC12 cells with protein kinase C did not decrease Nsp100 phosphorylation further. Moreover, preincubation of partially purified Nsp100 kinase with protein kinase C decreased its ability to phosphorylate Nsp100. These results suggest that the binding of nerve growth factor to its receptor on PC12 cells causes an increase in the activity of protein kinase C in the cytosol and phosphorylation of Nsp100 kinase, which in turn lowers its ability to phosphorylate Nsp100.

Full text

PDF
2353

Images in this article

2354Image
on p.2354

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Bocchini V., Angeletti P. U. The nerve growth factor: purification as a 30,000-molecular-weight protein. Proc Natl Acad Sci U S A. 1969 Oct;64(2):787–794. doi: 10.1073/pnas.64.2.787. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Burgess G. M., Godfrey P. P., McKinney J. S., Berridge M. J., Irvine R. F., Putney J. W., Jr The second messenger linking receptor activation to internal Ca release in liver. Nature. 1984 May 3;309(5963):63–66. doi: 10.1038/309063a0. [DOI] [PubMed] [Google Scholar]
  3. Burstein D. E., Greene L. A. Evidence for RNA synthesis-dependent and -independent pathways in stimulation of neurite outgrowth by nerve growth factor. Proc Natl Acad Sci U S A. 1978 Dec;75(12):6059–6063. doi: 10.1073/pnas.75.12.6059. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Cleveland D. W., Fischer S. G., Kirschner M. W., Laemmli U. K. Peptide mapping by limited proteolysis in sodium dodecyl sulfate and analysis by gel electrophoresis. J Biol Chem. 1977 Feb 10;252(3):1102–1106. [PubMed] [Google Scholar]
  5. Dichter M. A., Tischler A. S., Greene L. A. Nerve growth factor-induced increase in electrical excitability and acetylcholine sensitivity of a rat pheochromocytoma cell line. Nature. 1977 Aug 11;268(5620):501–504. doi: 10.1038/268501a0. [DOI] [PubMed] [Google Scholar]
  6. End D., Tolson N., Hashimoto S., Guroff G. Nerve growth factor-induced decrease in the cell-free phosphorylation of a soluble protein in PC12 cells. J Biol Chem. 1983 May 25;258(10):6549–6555. [PubMed] [Google Scholar]
  7. End D., Tolson N., Yu M. Y., Guroff G. Effects of 12-0-Tetradecanoylphorbol-13-acetate (TPA) on rat pheochromocytoma (PC12) cells: interactions with epidermal growth factor and nerve growth factor. J Cell Physiol. 1982 May;111(2):140–148. doi: 10.1002/jcp.1041110204. [DOI] [PubMed] [Google Scholar]
  8. Greene L. A., Rein G. Synthesis, storage and release of acetylcholine by a noradrenergic pheochromocytoma cell line. Nature. 1977 Jul 28;268(5618):349–351. doi: 10.1038/268349a0. [DOI] [PubMed] [Google Scholar]
  9. Greene L. A., Shooter E. M. The nerve growth factor: biochemistry, synthesis, and mechanism of action. Annu Rev Neurosci. 1980;3:353–402. doi: 10.1146/annurev.ne.03.030180.002033. [DOI] [PubMed] [Google Scholar]
  10. Greene L. A., Tischler A. S. Establishment of a noradrenergic clonal line of rat adrenal pheochromocytoma cells which respond to nerve growth factor. Proc Natl Acad Sci U S A. 1976 Jul;73(7):2424–2428. doi: 10.1073/pnas.73.7.2424. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Halegoua S., Patrick J. Nerve growth factor mediates phosphorylation of specific proteins. Cell. 1980 Nov;22(2 Pt 2):571–581. doi: 10.1016/0092-8674(80)90367-0. [DOI] [PubMed] [Google Scholar]
  12. Hama T., Guroff G. Distribution of Nsp100 and Nsp100 kinase, a nerve growth factor-sensitive phosphorylation system, in rat tissues. J Neurochem. 1985 Oct;45(4):1279–1287. doi: 10.1111/j.1471-4159.1985.tb05554.x. [DOI] [PubMed] [Google Scholar]
  13. Heumann R., Schwab M., Thoenen H. A second messenger required for nerve growth factor biological activity? Nature. 1981 Aug 27;292(5826):838–840. doi: 10.1038/292838a0. [DOI] [PubMed] [Google Scholar]
  14. Hsu L., Natyzak D., Laskin J. D. Effects of the tumor promoter 12-O-tetradecanoylphorbol-13-acetate on neurite outgrowth from chick embryo sensory ganglia. Cancer Res. 1984 Oct;44(10):4607–4614. [PubMed] [Google Scholar]
  15. Ishii D. N., Recio-Pinto E., Spinelli W., Mill J. F., Sonnenfeld K. H. Neurite formation modulated by nerve growth factor, insulin, and tumor promoter receptors. Int J Neurosci. 1985 Apr;26(1-2):109–127. doi: 10.3109/00207458508985610. [DOI] [PubMed] [Google Scholar]
  16. Joseph S. K., Thomas A. P., Williams R. J., Irvine R. F., Williamson J. R. myo-Inositol 1,4,5-trisphosphate. A second messenger for the hormonal mobilization of intracellular Ca2+ in liver. J Biol Chem. 1984 Mar 10;259(5):3077–3081. [PubMed] [Google Scholar]
  17. Joseph S. K., Williams R. J., Corkey B. E., Matschinsky F. M., Williamson J. R. The effect of inositol trisphosphate on Ca2+ fluxes in insulin-secreting tumor cells. J Biol Chem. 1984 Nov 10;259(21):12952–12955. [PubMed] [Google Scholar]
  18. Kikkawa U., Takai Y., Minakuchi R., Inohara S., Nishizuka Y. Calcium-activated, phospholipid-dependent protein kinase from rat brain. Subcellular distribution, purification, and properties. J Biol Chem. 1982 Nov 25;257(22):13341–13348. [PubMed] [Google Scholar]
  19. Koike T. Nerve growth factor-induced neurite outgrowth of rat pheochromocytoma PC 12 cells: dependence on extracellular Mg2+ and Ca2+. Brain Res. 1983 Dec 19;289(1-2):293–303. doi: 10.1016/0006-8993(83)90030-6. [DOI] [PubMed] [Google Scholar]
  20. LOWRY O. H., ROSEBROUGH N. J., FARR A. L., RANDALL R. J. Protein measurement with the Folin phenol reagent. J Biol Chem. 1951 Nov;193(1):265–275. [PubMed] [Google Scholar]
  21. Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
  22. Lakshmanan J. Nerve growth factor induced turnover of phosphatidylinositol in rat superior cervical ganglia. Biochem Biophys Res Commun. 1978 Jun 14;82(3):767–775. doi: 10.1016/0006-291x(78)90848-3. [DOI] [PubMed] [Google Scholar]
  23. Landreth G. E., Rieser G. D. Nerve growth factor- and epidermal growth factor-stimulated phosphorylation of a PC12 cytoskeletally associated protein in situ. J Cell Biol. 1985 Mar;100(3):677–683. doi: 10.1083/jcb.100.3.677. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Landreth G., Cohen P., Shooter E. M. Ca2+ transmembrane fluxes and nerve growth factor action on a clonal cell line of rat phaeochromocytoma. Nature. 1980 Jan 10;283(5743):202–204. doi: 10.1038/283202a0. [DOI] [PubMed] [Google Scholar]
  25. Levi-Montalcini R., Angeletti P. U. Nerve growth factor. Physiol Rev. 1968 Jul;48(3):534–569. doi: 10.1152/physrev.1968.48.3.534. [DOI] [PubMed] [Google Scholar]
  26. Levi A., Shechter Y., Neufeld E. J., Schlessinger J. Mobility, clustering, and transport of nerve growth factor in embryonal sensory cells and in a sympathetic neuronal cell line. Proc Natl Acad Sci U S A. 1980 Jun;77(6):3469–3473. doi: 10.1073/pnas.77.6.3469. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Nakanishi N., Guroff G. Nerve growth factor-induced increase in the cell-free phosphorylation of a nuclear protein in PC12 cells. J Biol Chem. 1985 Jun 25;260(12):7791–7799. [PubMed] [Google Scholar]
  28. Por S. B., Huttner W. B. A Mr 70,000 phosphoprotein of sympathetic neurons regulated by nerve growth factor and by depolarization. J Biol Chem. 1984 May 25;259(10):6526–6533. [PubMed] [Google Scholar]
  29. Prentki M., Biden T. J., Janjic D., Irvine R. F., Berridge M. J., Wollheim C. B. Rapid mobilization of Ca2+ from rat insulinoma microsomes by inositol-1,4,5-trisphosphate. Nature. 1984 Jun 7;309(5968):562–564. doi: 10.1038/309562a0. [DOI] [PubMed] [Google Scholar]
  30. Schubert D., LaCorbiere M., Whitlock C., Stallcup W. Alterations in the surface properties of cells responsive to nerve growth factor. Nature. 1978 Jun 29;273(5665):718–723. doi: 10.1038/273718a0. [DOI] [PubMed] [Google Scholar]
  31. Spinelli W., Ishii D. N. Tumor promoter receptors regulating neurite formation in cultured human neuroblastoma cells. Cancer Res. 1983 Sep;43(9):4119–4125. [PubMed] [Google Scholar]
  32. Streb H., Irvine R. F., Berridge M. J., Schulz I. Release of Ca2+ from a nonmitochondrial intracellular store in pancreatic acinar cells by inositol-1,4,5-trisphosphate. Nature. 1983 Nov 3;306(5938):67–69. doi: 10.1038/306067a0. [DOI] [PubMed] [Google Scholar]
  33. Thoenen H., Barde Y. A. Physiology of nerve growth factor. Physiol Rev. 1980 Oct;60(4):1284–1335. doi: 10.1152/physrev.1980.60.4.1284. [DOI] [PubMed] [Google Scholar]
  34. Togari A., Guroff G. Partial purification and characterization of a nerve growth factor-sensitive kinase and its substrate from PC12 cells. J Biol Chem. 1985 Mar 25;260(6):3804–3811. [PubMed] [Google Scholar]
  35. Traynor A. E. The relationship between neurite extension and phospholipid metabolism in PC12 cells. Brain Res. 1984 Jun;316(2):205–210. [PubMed] [Google Scholar]
  36. Vilgrain I., Cochet C., Chambaz E. M. Hormonal regulation of a calcium-activated, phospholipid-dependent protein kinase in bovine adrenal cortex. J Biol Chem. 1984 Mar 25;259(6):3403–3406. [PubMed] [Google Scholar]
  37. Weinstein I. B., Lee L. S., Fisher P. B., Mufson A., Yamasaki H. Action of phorbol esters in cell culture: mimicry of transformation, altered differentiation, and effects on cell membranes. J Supramol Struct. 1979;12(2):195–208. doi: 10.1002/jss.400120206. [DOI] [PubMed] [Google Scholar]
  38. Yu M. W., Tolson N. W., Guroff G. Increased phosphorylation of specific nuclear proteins in superior cervical ganglia and PC12 cells in response to nerve growth factor. J Biol Chem. 1980 Nov 10;255(21):10481–10492. [PubMed] [Google Scholar]

Articles from Proceedings of the National Academy of Sciences of the United States of America are provided here courtesy of National Academy of Sciences

RESOURCES