Skip to main content
PMC home page
Molecular Biology of the Cell logoLink to Molecular Biology of the Cell
. 1992 Mar;3(3):363–371. doi: 10.1091/mbc.3.3.363

Nerve growth factor employs multiple pathways to induce primary response genes in PC12 cells.

A Batistatou 1, C Volonté 1, L A Greene 1
PMCID: PMC275536  PMID: 1627834

Abstract

Nerve growth factor (NGF) leads to neuronal differentiation of PC12 cells and promotes their survival in serum-free medium. Past studies have shown that purine analogues block some of the effects of NGF but not others and thus that they can be used to dissect the mechanistic pathways of its action. In the present work we used 2-aminopurine (2-AP) and 6-thioguanine (6-TG) to examine whether NGF causes activation of primary response genes through a single signaling pathway or via multiple pathways. Northern blot analysis and nuclear run-off transcription assays were used to assess the activation of c-fos, c-jun, TIS1, TIS8, and TIS11 after exposure of PC12 cells to NGF in the presence or absence of 2-AP and 6-TG. Our findings indicate that NGF appears to employ at least three distinct pathways to induce early genes in PC12 cells. This suggests that the NGF signaling mechanism diverges at an early point after interaction of NGF with its receptor.

Full text

PDF
363

Images in this article

365Image
on p.365
365Image
on p.365
366Image
on p.366
366Image
on p.366
368Image
on p.368
368Image
on p.368
369Image
on p.369
369Image
on p.369

Selected References

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

  1. Altin J. G., Kujubu D. A., Raffioni S., Eveleth D. D., Herschman H. R., Bradshaw R. A. Differential induction of primary-response (TIS) genes in PC12 pheochromocytoma cells and the unresponsive variant PC12nnr5. J Biol Chem. 1991 Mar 25;266(9):5401–5406. [PubMed] [Google Scholar]
  2. Bohmann D., Bos T. J., Admon A., Nishimura T., Vogt P. K., Tjian R. Human proto-oncogene c-jun encodes a DNA binding protein with structural and functional properties of transcription factor AP-1. Science. 1987 Dec 4;238(4832):1386–1392. doi: 10.1126/science.2825349. [DOI] [PubMed] [Google Scholar]
  3. Boulton T. G., Gregory J. S., Cobb M. H. Purification and properties of extracellular signal-regulated kinase 1, an insulin-stimulated microtubule-associated protein 2 kinase. Biochemistry. 1991 Jan 8;30(1):278–286. doi: 10.1021/bi00215a038. [DOI] [PubMed] [Google Scholar]
  4. Cho K. O., Skarnes W. C., Minsk B., Palmieri S., Jackson-Grusby L., Wagner J. A. Nerve growth factor regulates gene expression by several distinct mechanisms. Mol Cell Biol. 1989 Jan;9(1):135–143. doi: 10.1128/mcb.9.1.135. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Chomczynski P., Sacchi N. Single-step method of RNA isolation by acid guanidinium thiocyanate-phenol-chloroform extraction. Anal Biochem. 1987 Apr;162(1):156–159. doi: 10.1006/abio.1987.9999. [DOI] [PubMed] [Google Scholar]
  6. Curran T., Peters G., Van Beveren C., Teich N. M., Verma I. M. FBJ murine osteosarcoma virus: identification and molecular cloning of biologically active proviral DNA. J Virol. 1982 Nov;44(2):674–682. doi: 10.1128/jvi.44.2.674-682.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Farrell P. J., Balkow K., Hunt T., Jackson R. J., Trachsel H. Phosphorylation of initiation factor elF-2 and the control of reticulocyte protein synthesis. Cell. 1977 May;11(1):187–200. doi: 10.1016/0092-8674(77)90330-0. [DOI] [PubMed] [Google Scholar]
  8. Greenberg M. E., Greene L. A., Ziff E. B. Nerve growth factor and epidermal growth factor induce rapid transient changes in proto-oncogene transcription in PC12 cells. J Biol Chem. 1985 Nov 15;260(26):14101–14110. [PubMed] [Google Scholar]
  9. Greene L. A. Nerve growth factor prevents the death and stimulates the neuronal differentiation of clonal PC12 pheochromocytoma cells in serum-free medium. J Cell Biol. 1978 Sep;78(3):747–755. doi: 10.1083/jcb.78.3.747. [DOI] [PMC free article] [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. Greene L. A., Volonté C., Chalazonitis A. Purine analogs inhibit nerve growth factor-promoted neurite outgrowth by sympathetic and sensory neurons. J Neurosci. 1990 May;10(5):1479–1485. doi: 10.1523/JNEUROSCI.10-05-01479.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Halazonetis T. D., Georgopoulos K., Greenberg M. E., Leder P. c-Jun dimerizes with itself and with c-Fos, forming complexes of different DNA binding affinities. Cell. 1988 Dec 2;55(5):917–924. doi: 10.1016/0092-8674(88)90147-x. [DOI] [PubMed] [Google Scholar]
  13. Herschman H. R. Primary response genes induced by growth factors and tumor promoters. Annu Rev Biochem. 1991;60:281–319. doi: 10.1146/annurev.bi.60.070191.001433. [DOI] [PubMed] [Google Scholar]
  14. Kaplan D. R., Hempstead B. L., Martin-Zanca D., Chao M. V., Parada L. F. The trk proto-oncogene product: a signal transducing receptor for nerve growth factor. Science. 1991 Apr 26;252(5005):554–558. doi: 10.1126/science.1850549. [DOI] [PubMed] [Google Scholar]
  15. Kujubu D. A., Lim R. W., Varnum B. C., Herschman H. R. Induction of transiently expressed genes in PC-12 pheochromocytoma cells. Oncogene. 1987;1(3):257–262. [PubMed] [Google Scholar]
  16. 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]
  17. 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]
  18. Levi-Montalcini R. The nerve growth factor 35 years later. Science. 1987 Sep 4;237(4819):1154–1162. doi: 10.1126/science.3306916. [DOI] [PubMed] [Google Scholar]
  19. Levi A., Alemà S. The mechanism of action of nerve growth factor. Annu Rev Pharmacol Toxicol. 1991;31:205–228. doi: 10.1146/annurev.pa.31.040191.001225. [DOI] [PubMed] [Google Scholar]
  20. Lim R. W., Varnum B. C., Herschman H. R. Cloning of tetradecanoyl phorbol ester-induced 'primary response' sequences and their expression in density-arrested Swiss 3T3 cells and a TPA non-proliferative variant. Oncogene. 1987;1(3):263–270. [PubMed] [Google Scholar]
  21. Loeb D. M., Maragos J., Martin-Zanca D., Chao M. V., Parada L. F., Greene L. A. The trk proto-oncogene rescues NGF responsiveness in mutant NGF-nonresponsive PC12 cell lines. Cell. 1991 Sep 6;66(5):961–966. doi: 10.1016/0092-8674(91)90441-z. [DOI] [PubMed] [Google Scholar]
  22. Machida C. M., Scott J. D., Ciment G. NGF-induction of the metalloproteinase-transin/stromelysin in PC12 cells: involvement of multiple protein kinases. J Cell Biol. 1991 Sep;114(5):1037–1048. doi: 10.1083/jcb.114.5.1037. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Mahadevan L. C., Wills A. J., Hirst E. A., Rathjen P. D., Heath J. K. 2-Aminopurine abolishes EGF- and TPA-stimulated pp33 phosphorylation and c-fos induction without affecting the activation of protein kinase C. Oncogene. 1990 Mar;5(3):327–335. [PubMed] [Google Scholar]
  24. Matsuda Y., Guroff G. Purification and mechanism of activation of a nerve growth factor-sensitive S6 kinase from PC12 cells. J Biol Chem. 1987 Feb 25;262(6):2832–2844. [PubMed] [Google Scholar]
  25. Milbrandt J. A nerve growth factor-induced gene encodes a possible transcriptional regulatory factor. Science. 1987 Nov 6;238(4828):797–799. doi: 10.1126/science.3672127. [DOI] [PubMed] [Google Scholar]
  26. Milbrandt J. Nerve growth factor induces a gene homologous to the glucocorticoid receptor gene. Neuron. 1988 May;1(3):183–188. doi: 10.1016/0896-6273(88)90138-9. [DOI] [PubMed] [Google Scholar]
  27. Milbrandt J. Nerve growth factor rapidly induces c-fos mRNA in PC12 rat pheochromocytoma cells. Proc Natl Acad Sci U S A. 1986 Jul;83(13):4789–4793. doi: 10.1073/pnas.83.13.4789. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Mobley W. C., Schenker A., Shooter E. M. Characterization and isolation of proteolytically modified nerve growth factor. Biochemistry. 1976 Dec 14;15(25):5543–5552. doi: 10.1021/bi00670a019. [DOI] [PubMed] [Google Scholar]
  29. Piechaczyk M., Blanchard J. M., Marty L., Dani C., Panabieres F., El Sabouty S., Fort P., Jeanteur P. Post-transcriptional regulation of glyceraldehyde-3-phosphate-dehydrogenase gene expression in rat tissues. Nucleic Acids Res. 1984 Sep 25;12(18):6951–6963. doi: 10.1093/nar/12.18.6951. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Rowland E. A., Müller T. H., Goldstein M., Greene L. A. Cell-free detection and characterization of a novel nerve growth factor-activated protein kinase in PC12 cells. J Biol Chem. 1987 Jun 5;262(16):7504–7513. [PubMed] [Google Scholar]
  31. Sheng M., Thompson M. A., Greenberg M. E. CREB: a Ca(2+)-regulated transcription factor phosphorylated by calmodulin-dependent kinases. Science. 1991 Jun 7;252(5011):1427–1430. doi: 10.1126/science.1646483. [DOI] [PubMed] [Google Scholar]
  32. Tsao H., Aletta J. M., Greene L. A. Nerve growth factor and fibroblast growth factor selectively activate a protein kinase that phosphorylates high molecular weight microtubule-associated proteins. Detection, partial purification, and characterization in PC12 cells. J Biol Chem. 1990 Sep 15;265(26):15471–15480. [PubMed] [Google Scholar]
  33. Varnum B. C., Lim R. W., Sukhatme V. P., Herschman H. R. Nucleotide sequence of a cDNA encoding TIS11, a message induced in Swiss 3T3 cells by the tumor promoter tetradecanoyl phorbol acetate. Oncogene. 1989 Jan;4(1):119–120. [PubMed] [Google Scholar]
  34. Volonté C., Greene L. A. 6-Methylmercaptopurine riboside is a potent and selective inhibitor of nerve growth factor-activated protein kinase N. J Neurochem. 1992 Feb;58(2):700–708. doi: 10.1111/j.1471-4159.1992.tb09774.x. [DOI] [PubMed] [Google Scholar]
  35. Volonté C., Greene L. A. Induction of ornithine decarboxylase by nerve growth factor in PC12 cells: dissection by purine analogues. J Biol Chem. 1990 Jul 5;265(19):11050–11055. [PubMed] [Google Scholar]
  36. Volonté C., Rukenstein A., Loeb D. M., Greene L. A. Differential inhibition of nerve growth factor responses by purine analogues: correlation with inhibition of a nerve growth factor-activated protein kinase. J Cell Biol. 1989 Nov;109(5):2395–2403. doi: 10.1083/jcb.109.5.2395. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. Wu B. Y., Fodor E. J., Edwards R. H., Rutter W. J. Nerve growth factor induces the proto-oncogene c-jun in PC12 cells. J Biol Chem. 1989 May 25;264(15):9000–9003. [PubMed] [Google Scholar]
  38. Zinn K., Keller A., Whittemore L. A., Maniatis T. 2-Aminopurine selectively inhibits the induction of beta-interferon, c-fos, and c-myc gene expression. Science. 1988 Apr 8;240(4849):210–213. doi: 10.1126/science.3281258. [DOI] [PubMed] [Google Scholar]

Articles from Molecular Biology of the Cell are provided here courtesy of American Society for Cell Biology

RESOURCES