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. 1988 Jan 1;106(1):181–193. doi: 10.1083/jcb.106.1.181

A nerve growth factor-regulated messenger RNA encodes a new intermediate filament protein

PMCID: PMC2114951  PMID: 3339087

Abstract

Differential screening of a cDNA library from the PC12 rat pheochromocytoma cell line previously revealed a clone, clone 73, whose corresponding mRNA is induced by nerve growth factor (NGF). Induction parallels NGF-stimulated PC12 differentiation from a chromaffinlike phenotype to a sympathetic neuronlike phenotype. We report that DNA sequence analysis reveals that clone 73 mRNA encodes an intermediate filament (IF) protein whose predicted amino acid sequence is distinct from the known sequences of other members of the IF protein family. The sequence has highest homology with desmin and vimentin and includes the highly conserved central alpha-helical rod domain with the characteristic heptad repeat of hydrophobic residues, but has lower homology in the amino-terminal head and carboxyl-terminal tail domains. The head domain contains a large number of serine residues which are potential phosphorylation sites. The expression of clone 73 in vivo in the nervous system of the adult rat was investigated by in situ hybridization of clone 73 probes to tissue sections. The mRNA is expressed at high levels in ganglia of the peripheral nervous system, including the superior cervical ganglion (sympathetic), ciliary ganglion (parasympathetic), and dorsal root ganglion (sensory). In the central nervous system, motor nuclei of cranial nerves III, IV, V, VI, VII, X, and XII as well as ventral horn motor neurons and a restricted set of other central nervous system nuclei express the clone 73 mRNA. Tissues apart from those of the nervous system did not in general express the mRNA, with only very low levels detected in adrenal gland. We discuss the implications of these results for the mechanism of NGF- induced PC12 cell differentiation, the pathways of neuronal development in vivo, and the possible function of the clone 73 IF protein and its relationship to other IF proteins.

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Selected References

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  1. Aletta J. M., Greene L. A. Sequential phosphorylation of chartin microtubule-associated proteins is regulated by the presence of microtubules. J Cell Biol. 1987 Jul;105(1):277–290. doi: 10.1083/jcb.105.1.277. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Anderson D. J., Axel R. A bipotential neuroendocrine precursor whose choice of cell fate is determined by NGF and glucocorticoids. Cell. 1986 Dec 26;47(6):1079–1090. doi: 10.1016/0092-8674(86)90823-8. [DOI] [PubMed] [Google Scholar]
  3. Anderson D. J., Axel R. Molecular probes for the development and plasticity of neural crest derivatives. Cell. 1985 Sep;42(2):649–662. doi: 10.1016/0092-8674(85)90122-9. [DOI] [PubMed] [Google Scholar]
  4. Biggin M. D., Gibson T. J., Hong G. F. Buffer gradient gels and 35S label as an aid to rapid DNA sequence determination. Proc Natl Acad Sci U S A. 1983 Jul;80(13):3963–3965. doi: 10.1073/pnas.80.13.3963. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Bjerre B., Björklund A. The production of catecholamine-containing cells in vitro by young chick embryos: effects of "nerve growth factor" (NGF) and its antiserum. Neurobiology. 1973;3(3):140–161. [PubMed] [Google Scholar]
  6. Collins F., Dawson A. An effect of nerve growth factor on parasympathetic neurite outgrowth. Proc Natl Acad Sci U S A. 1983 Apr;80(7):2091–2094. doi: 10.1073/pnas.80.7.2091. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Cox K. H., DeLeon D. V., Angerer L. M., Angerer R. C. Detection of mrnas in sea urchin embryos by in situ hybridization using asymmetric RNA probes. Dev Biol. 1984 Feb;101(2):485–502. doi: 10.1016/0012-1606(84)90162-3. [DOI] [PubMed] [Google Scholar]
  8. Doupe A. J., Landis S. C., Patterson P. H. Environmental influences in the development of neural crest derivatives: glucocorticoids, growth factors, and chromaffin cell plasticity. J Neurosci. 1985 Aug;5(8):2119–2142. doi: 10.1523/JNEUROSCI.05-08-02119.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Doupe A. J., Patterson P. H., Landis S. C. Small intensely fluorescent cells in culture: role of glucocorticoids and growth factors in their development and interconversions with other neural crest derivatives. J Neurosci. 1985 Aug;5(8):2143–2160. doi: 10.1523/JNEUROSCI.05-08-02143.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Fisher D. Z., Chaudhary N., Blobel G. cDNA sequencing of nuclear lamins A and C reveals primary and secondary structural homology to intermediate filament proteins. Proc Natl Acad Sci U S A. 1986 Sep;83(17):6450–6454. doi: 10.1073/pnas.83.17.6450. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Franke W. W., Grund C., Achtstätter T. Co-expression of cytokeratins and neurofilament proteins in a permanent cell line: cultured rat PC12 cells combine neuronal and epithelial features. J Cell Biol. 1986 Nov;103(5):1933–1943. doi: 10.1083/jcb.103.5.1933. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Korsching S., Auburger G., Heumann R., Scott J., Thoenen H. Levels of nerve growth factor and its mRNA in the central nervous system of the rat correlate with cholinergic innervation. EMBO J. 1985 Jun;4(6):1389–1393. doi: 10.1002/j.1460-2075.1985.tb03791.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Krieg T. M., Schafer M. P., Cheng C. K., Filpula D., Flaherty P., Steinert P. M., Roop D. R. Organization of a type I keratin gene. Evidence for evolution of intermediate filaments from a common ancestral gene. J Biol Chem. 1985 May 25;260(10):5867–5870. [PubMed] [Google Scholar]
  14. Lazarides E. Intermediate filaments: a chemically heterogeneous, developmentally regulated class of proteins. Annu Rev Biochem. 1982;51:219–250. doi: 10.1146/annurev.bi.51.070182.001251. [DOI] [PubMed] [Google Scholar]
  15. 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]
  16. 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]
  17. 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]
  18. Lewis S. A., Balcarek J. M., Krek V., Shelanski M., Cowan N. J. Sequence of a cDNA clone encoding mouse glial fibrillary acidic protein: structural conservation of intermediate filaments. Proc Natl Acad Sci U S A. 1984 May;81(9):2743–2746. doi: 10.1073/pnas.81.9.2743. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Lewis S. A., Cowan N. J. Anomalous placement of introns in a member of the intermediate filament multigene family: an evolutionary conundrum. Mol Cell Biol. 1986 May;6(5):1529–1534. doi: 10.1128/mcb.6.5.1529. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Liem R. K., Yen S. H., Salomon G. D., Shelanski M. L. Intermediate filaments in nervous tissues. J Cell Biol. 1978 Dec;79(3):637–645. doi: 10.1083/jcb.79.3.637. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. McKeon F. D., Kirschner M. W., Caput D. Homologies in both primary and secondary structure between nuclear envelope and intermediate filament proteins. Nature. 1986 Feb 6;319(6053):463–468. doi: 10.1038/319463a0. [DOI] [PubMed] [Google Scholar]
  22. McLachlan A. D., Stewart M. Tropomyosin coiled-coil interactions: evidence for an unstaggered structure. J Mol Biol. 1975 Oct 25;98(2):293–304. doi: 10.1016/s0022-2836(75)80119-7. [DOI] [PubMed] [Google Scholar]
  23. Messing J., Vieira J. A new pair of M13 vectors for selecting either DNA strand of double-digest restriction fragments. Gene. 1982 Oct;19(3):269–276. doi: 10.1016/0378-1119(82)90016-6. [DOI] [PubMed] [Google Scholar]
  24. Mobley W. C., Rutkowski J. L., Tennekoon G. I., Buchanan K., Johnston M. V. Choline acetyltransferase activity in striatum of neonatal rats increased by nerve growth factor. Science. 1985 Jul 19;229(4710):284–287. doi: 10.1126/science.2861660. [DOI] [PubMed] [Google Scholar]
  25. Napolitano E. W., Chin S. S., Colman D. R., Liem R. K. Complete amino acid sequence and in vitro expression of rat NF-M, the middle molecular weight neurofilament protein. J Neurosci. 1987 Aug;7(8):2590–2599. [PMC free article] [PubMed] [Google Scholar]
  26. Parysek L. M., Goldman R. D. Characterization of intermediate filaments in PC12 cells. J Neurosci. 1987 Mar;7(3):781–791. doi: 10.1523/JNEUROSCI.07-03-00781.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Portier M. M., Brachet P., Croizat B., Gros F. Regulation of peripherin in mouse neuroblastoma and rat PC 12 pheochromocytoma cell lines. Dev Neurosci. 1983;6(4-5):215–226. doi: 10.1159/000112348. [DOI] [PubMed] [Google Scholar]
  28. Portier M. M., de Néchaud B., Gros F. Peripherin, a new member of the intermediate filament protein family. Dev Neurosci. 1983;6(6):335–344. doi: 10.1159/000112360. [DOI] [PubMed] [Google Scholar]
  29. Quax W., Egberts W. V., Hendriks W., Quax-Jeuken Y., Bloemendal H. The structure of the vimentin gene. Cell. 1983 Nov;35(1):215–223. doi: 10.1016/0092-8674(83)90224-6. [DOI] [PubMed] [Google Scholar]
  30. Quax W., van den Broek L., Egberts W. V., Ramaekers F., Bloemendal H. Characterization of the hamster desmin gene: expression and formation of desmin filaments in nonmuscle cells after gene transfer. Cell. 1985 Nov;43(1):327–338. doi: 10.1016/0092-8674(85)90038-8. [DOI] [PubMed] [Google Scholar]
  31. Raivich G., Zimmermann A., Sutter A. The spatial and temporal pattern of beta NGF receptor expression in the developing chick embryo. EMBO J. 1985 Mar;4(3):637–644. doi: 10.1002/j.1460-2075.1985.tb03677.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Richardson P. M., Issa V. M., Riopelle R. J. Distribution of neuronal receptors for nerve growth factor in the rat. J Neurosci. 1986 Aug;6(8):2312–2321. doi: 10.1523/JNEUROSCI.06-08-02312.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Rydel R. E., Greene L. A. Acidic and basic fibroblast growth factors promote stable neurite outgrowth and neuronal differentiation in cultures of PC12 cells. J Neurosci. 1987 Nov;7(11):3639–3653. doi: 10.1523/JNEUROSCI.07-11-03639.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Sanger F., Coulson A. R., Barrell B. G., Smith A. J., Roe B. A. Cloning in single-stranded bacteriophage as an aid to rapid DNA sequencing. J Mol Biol. 1980 Oct 25;143(2):161–178. doi: 10.1016/0022-2836(80)90196-5. [DOI] [PubMed] [Google Scholar]
  35. Sanger F., Coulson A. R. The use of thin acrylamide gels for DNA sequencing. FEBS Lett. 1978 Mar 1;87(1):107–110. doi: 10.1016/0014-5793(78)80145-8. [DOI] [PubMed] [Google Scholar]
  36. Sanger F., Nicklen S., Coulson A. R. DNA sequencing with chain-terminating inhibitors. Proc Natl Acad Sci U S A. 1977 Dec;74(12):5463–5467. doi: 10.1073/pnas.74.12.5463. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. Steinert P. M., Parry D. A., Idler W. W., Johnson L. D., Steven A. C., Roop D. R. Amino acid sequences of mouse and human epidermal type II keratins of Mr 67,000 provide a systematic basis for the structural and functional diversity of the end domains of keratin intermediate filament subunits. J Biol Chem. 1985 Jun 10;260(11):7142–7149. [PubMed] [Google Scholar]
  38. Steinert P. M., Steven A. C., Roop D. R. The molecular biology of intermediate filaments. Cell. 1985 Sep;42(2):411–420. doi: 10.1016/0092-8674(85)90098-4. [DOI] [PubMed] [Google Scholar]
  39. 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]
  40. Togari A., Dickens G., Kuzuya H., Guroff G. The effect of fibroblast growth factor on PC12 cells. J Neurosci. 1985 Feb;5(2):307–316. doi: 10.1523/JNEUROSCI.05-02-00307.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  41. Trojanowski J. Q., Walkenstein N., Lee V. M. Expression of neurofilament subunits in neurons of the central and peripheral nervous system: an immunohistochemical study with monoclonal antibodies. J Neurosci. 1986 Mar;6(3):650–660. doi: 10.1523/JNEUROSCI.06-03-00650.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  42. Weber K., Geisler N. Intermediate filaments: structural conservation and divergence. Ann N Y Acad Sci. 1985;455:126–143. doi: 10.1111/j.1749-6632.1985.tb50408.x. [DOI] [PubMed] [Google Scholar]

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