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. 1989 Jan;9(1):135–143. doi: 10.1128/mcb.9.1.135

Nerve growth factor regulates gene expression by several distinct mechanisms.

K O Cho 1, W C Skarnes 1, B Minsk 1, S Palmieri 1, L Jackson-Grusby 1, J A Wagner 1
PMCID: PMC362154  PMID: 2538715

Abstract

To help elucidate the mechanisms by which nerve growth factor (NGF) regulates gene expression, we have identified and studied four genes (a-2, d-2, d-4, and d-5) that are positively regulated by NGF in PC12 cells, including one (d-2) which has previously been identified as a putative transcription factor (NGF I-A). Three of these genes, including d-2, were induced very rapidly at the transcriptional level, but the relative time courses of transcription and mRNA accumulation of each of these three genes were distinct. The fourth gene (d-4) displayed no apparent increase in transcription that corresponded to the increase in its mRNA, suggesting that NGF may regulate its expression at a posttranscriptional level. Thus, NGF positively regulates gene expression by more than one mechanism. These genes could also be distinguished on the basis of their response to cyclic AMP. The expression of d-2 and a-2 was increased by cholera toxin and further augmented by NGF; however, cholera toxin not only failed to increase the levels of d-5 and d-4 mRNA but also actually inhibited the NGF-dependent increase. The expression of each of these genes, including d-2 (NGF I-A), was also increased by fibroblast growth factor, epidermal growth factor (EGF), phorbol myristate acetate, and in some cases insulin, showing that the regulation of these genes is not unique to NGF. Because each of these genes was expressed in response to phorbol myristate acetate and EGF, their expression may be necessary but is certainly not sufficient for neurite formation. The protein kinase inhibitor K-252a prevented the NGF-associated, but not the acidic FGF-associated, induction of d-2 and d-5 gene expression, suggesting that these two growth factors may regulate gene expression via different cellular pathways. The study of the regulation of the expression of these and other NGF-inducible genes should valuable new information concerning how NGF and other growth factors cause neural differentiation.

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Selected References

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

  1. Benton W. D., Davis R. W. Screening lambdagt recombinant clones by hybridization to single plaques in situ. Science. 1977 Apr 8;196(4286):180–182. doi: 10.1126/science.322279. [DOI] [PubMed] [Google Scholar]
  2. Burstein D. E., Blumberg P. M., Greene L. A. Nerve growth factor-induced neuronal differentiation of PC12 pheochromocytoma cells: lack of inhibition by a tumor promoter. Brain Res. 1982 Sep 9;247(1):115–119. doi: 10.1016/0006-8993(82)91033-2. [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. Chirgwin J. M., Przybyla A. E., MacDonald R. J., Rutter W. J. Isolation of biologically active ribonucleic acid from sources enriched in ribonuclease. Biochemistry. 1979 Nov 27;18(24):5294–5299. doi: 10.1021/bi00591a005. [DOI] [PubMed] [Google Scholar]
  5. Comb M., Birnberg N. C., Seasholtz A., Herbert E., Goodman H. M. A cyclic AMP- and phorbol ester-inducible DNA element. 1986 Sep 25-Oct 1Nature. 323(6086):353–356. doi: 10.1038/323353a0. [DOI] [PubMed] [Google Scholar]
  6. Contreras M. L., Guroff G. Calcium-dependent nerve growth factor-stimulated hydrolysis of phosphoinositides in PC12 cells. J Neurochem. 1987 May;48(5):1466–1472. doi: 10.1111/j.1471-4159.1987.tb05687.x. [DOI] [PubMed] [Google Scholar]
  7. Cremins J., Wagner J. A., Halegoua S. Nerve growth factor action is mediated by cyclic AMP- and Ca+2/phospholipid-dependent protein kinases. J Cell Biol. 1986 Sep;103(3):887–893. doi: 10.1083/jcb.103.3.887. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. 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]
  9. D'Amore P. A., Klagsbrun M. Endothelial cell mitogens derived from retina and hypothalamus: biochemical and biological similarities. J Cell Biol. 1984 Oct;99(4 Pt 1):1545–1549. doi: 10.1083/jcb.99.4.1545. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. 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]
  11. Doherty P., Mann D. A., Walsh F. S. Cholera toxin and dibutyryl cyclic AMP inhibit the expression of neurofilament protein induced by nerve growth factor in cultures of naive and primed PC12 cells. J Neurochem. 1987 Dec;49(6):1676–1687. doi: 10.1111/j.1471-4159.1987.tb02425.x. [DOI] [PubMed] [Google Scholar]
  12. Drubin D. G., Feinstein S. C., Shooter E. M., Kirschner M. W. Nerve growth factor-induced neurite outgrowth in PC12 cells involves the coordinate induction of microtubule assembly and assembly-promoting factors. J Cell Biol. 1985 Nov;101(5 Pt 1):1799–1807. doi: 10.1083/jcb.101.5.1799. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Feinberg A. P., Vogelstein B. A technique for radiolabeling DNA restriction endonuclease fragments to high specific activity. Anal Biochem. 1983 Jul 1;132(1):6–13. doi: 10.1016/0003-2697(83)90418-9. [DOI] [PubMed] [Google Scholar]
  14. 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]
  15. Greenberg M. E., Siegfried Z., Ziff E. B. Mutation of the c-fos gene dyad symmetry element inhibits serum inducibility of transcription in vivo and the nuclear regulatory factor binding in vitro. Mol Cell Biol. 1987 Mar;7(3):1217–1225. doi: 10.1128/mcb.7.3.1217. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Greene L. A., Drexler S. A., Connolly J. L., Rukenstein A., Green S. H. Selective inhibition of responses to nerve growth factor and of microtubule-associated protein phosphorylation by activators of adenylate cyclase. J Cell Biol. 1986 Nov;103(5):1967–1978. doi: 10.1083/jcb.103.5.1967. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. 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]
  18. Gubler U., Hoffman B. J. A simple and very efficient method for generating cDNA libraries. Gene. 1983 Nov;25(2-3):263–269. doi: 10.1016/0378-1119(83)90230-5. [DOI] [PubMed] [Google Scholar]
  19. Gunning P. W., Landreth G. E., Bothwell M. A., Shooter E. M. Differential and synergistic actions of nerve growth factor and cyclic AMP in PC12 cells. J Cell Biol. 1981 May;89(2):240–245. doi: 10.1083/jcb.89.2.240. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. 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]
  21. Hama T., Huang K. P., Guroff G. Protein kinase C as a component of a nerve growth factor-sensitive phosphorylation system in PC12 cells. Proc Natl Acad Sci U S A. 1986 Apr;83(8):2353–2357. doi: 10.1073/pnas.83.8.2353. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Huff K. R., Guroff G. Nerve growth factor-induced reduction in epidermal growth factor responsiveness and epidermal growth factor receptors in PC12 cells: an aspect of cell differentiation. Biochem Biophys Res Commun. 1979 Jul 12;89(1):175–180. doi: 10.1016/0006-291x(79)90960-4. [DOI] [PubMed] [Google Scholar]
  23. Huffaker T., Corcoran T., Wagner J. A. Adenosine inhibits cell division and promotes neurite extension in PC12 cells. J Cell Physiol. 1984 Aug;120(2):188–196. doi: 10.1002/jcp.1041200212. [DOI] [PubMed] [Google Scholar]
  24. Karns L. R., Ng S. C., Freeman J. A., Fishman M. C. Cloning of complementary DNA for GAP-43, a neuronal growth-related protein. Science. 1987 May 1;236(4801):597–600. doi: 10.1126/science.2437653. [DOI] [PubMed] [Google Scholar]
  25. Koizumi S., Contreras M. L., Matsuda Y., Hama T., Lazarovici P., Guroff G. K-252a: a specific inhibitor of the action of nerve growth factor on PC 12 cells. J Neurosci. 1988 Feb;8(2):715–721. doi: 10.1523/JNEUROSCI.08-02-00715.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Kruijer W., Schubert D., Verma I. M. Induction of the proto-oncogene fos by nerve growth factor. Proc Natl Acad Sci U S A. 1985 Nov;82(21):7330–7334. doi: 10.1073/pnas.82.21.7330. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Leonard D. G., Ziff E. B., Greene L. A. Identification and characterization of mRNAs regulated by nerve growth factor in PC12 cells. Mol Cell Biol. 1987 Sep;7(9):3156–3167. doi: 10.1128/mcb.7.9.3156. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Levi-Montalcini R. The nerve growth factor: its mode of action on sensory and sympathetic nerve cells. Harvey Lect. 1966;60:217–259. [PubMed] [Google Scholar]
  29. Levi A., Eldridge J. D., Paterson B. M. Molecular cloning of a gene sequence regulated by nerve growth factor. Science. 1985 Jul 26;229(4711):393–395. doi: 10.1126/science.3839317. [DOI] [PubMed] [Google Scholar]
  30. Lindenbaum M. H., Carbonetto S., Grosveld F., Flavell D., Mushynski W. E. Transcriptional and post-transcriptional effects of nerve growth factor on expression of the three neurofilament subunits in PC-12 cells. J Biol Chem. 1988 Apr 25;263(12):5662–5667. [PubMed] [Google Scholar]
  31. Masiakowski P., Shooter E. M. Nerve growth factor induces the genes for two proteins related to a family of calcium-binding proteins in PC12 cells. Proc Natl Acad Sci U S A. 1988 Feb;85(4):1277–1281. doi: 10.1073/pnas.85.4.1277. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. 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]
  33. 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]
  34. 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]
  35. Prywes R., Roeder R. G. Inducible binding of a factor to the c-fos enhancer. Cell. 1986 Dec 5;47(5):777–784. doi: 10.1016/0092-8674(86)90520-9. [DOI] [PubMed] [Google Scholar]
  36. Richter-Landsberg C., Jastorff B. The role of cAMP in nerve growth factor-promoted neurite outgrowth in PC12 cells. J Cell Biol. 1986 Mar;102(3):821–829. doi: 10.1083/jcb.102.3.821. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. Sajovic P., Ennulat D. J., Shelanski M. L., Greene L. A. Isolation of NILE glycoprotein-related cDNA probes. J Neurochem. 1987 Sep;49(3):756–763. doi: 10.1111/j.1471-4159.1987.tb00958.x. [DOI] [PubMed] [Google Scholar]
  38. 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]
  39. Stein R., Orit S., Anderson D. J. The induction of a neural-specific gene, SCG10, by nerve growth factor in PC12 cells is transcriptional, protein synthesis dependent, and glucocorticoid inhibitable. Dev Biol. 1988 Jun;127(2):316–325. doi: 10.1016/0012-1606(88)90318-1. [DOI] [PubMed] [Google Scholar]
  40. Stumpo D. J., Stewart T. N., Gilman M. Z., Blackshear P. J. Identification of c-fos sequences involved in induction by insulin and phorbol esters. J Biol Chem. 1988 Feb 5;263(4):1611–1614. [PubMed] [Google Scholar]
  41. Suissa M. Spectrophotometric quantitation of silver grains eluted from autoradiograms. Anal Biochem. 1983 Sep;133(2):511–514. doi: 10.1016/0003-2697(83)90117-3. [DOI] [PubMed] [Google Scholar]
  42. 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]
  43. Togari A., Baker D., Dickens G., Guroff G. The neurite-promoting effect of fibroblast growth factor on PC12 cells. Biochem Biophys Res Commun. 1983 Aug 12;114(3):1189–1193. doi: 10.1016/0006-291x(83)90688-5. [DOI] [PubMed] [Google Scholar]
  44. Traynor A. E., Schubert D., Allen W. R. Alterations of lipid metabolism in response to nerve growth factor. J Neurochem. 1982 Dec;39(6):1677–1683. doi: 10.1111/j.1471-4159.1982.tb08002.x. [DOI] [PubMed] [Google Scholar]
  45. 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]
  46. Van Buskirk R., Corcoran T., Wagner J. A. Clonal variants of PC12 pheochromocytoma cells with defects in cAMP-dependent protein kinases induce ornithine decarboxylase in response to nerve growth factor but not to adenosine agonists. Mol Cell Biol. 1985 Aug;5(8):1984–1992. doi: 10.1128/mcb.5.8.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  47. Wagner J. A., D'Amore P. A. Neurite outgrowth induced by an endothelial cell mitogen isolated from retina. J Cell Biol. 1986 Oct;103(4):1363–1367. doi: 10.1083/jcb.103.4.1363. [DOI] [PMC free article] [PubMed] [Google Scholar]
  48. Williams L. R., Varon S., Peterson G. M., Wictorin K., Fischer W., Bjorklund A., Gage F. H. Continuous infusion of nerve growth factor prevents basal forebrain neuronal death after fimbria fornix transection. Proc Natl Acad Sci U S A. 1986 Dec;83(23):9231–9235. doi: 10.1073/pnas.83.23.9231. [DOI] [PMC free article] [PubMed] [Google Scholar]
  49. Yanisch-Perron C., Vieira J., Messing J. Improved M13 phage cloning vectors and host strains: nucleotide sequences of the M13mp18 and pUC19 vectors. Gene. 1985;33(1):103–119. doi: 10.1016/0378-1119(85)90120-9. [DOI] [PubMed] [Google Scholar]
  50. Yu M. W., Hori S., Tolson N., Huff K., Guroff G. Increased phosphorylation of a specific nuclear protein in rat superior cervical ganglia in response to nerve growth factor. Biochem Biophys Res Commun. 1978 Apr 14;81(3):941–946. doi: 10.1016/0006-291x(78)91442-0. [DOI] [PubMed] [Google Scholar]

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