Calcified Tissue International

, Volume 54, Issue 5, pp 426–430 | Cite as

The proto-oncogene C-raf-1 is highly expressed only in the hypertrophic zone of the growth plate

  • Y. Kaneko
  • H. Tanzawa
  • K. Sato
Molecular and Cellular Biology


Proto-oncogene c-raf-1, the cellular homologue of the acutely transforming oncogene v-raf, has a central role in the signal tranduction pathways. The growth plate, due to its non-overlapping zones of chondrocyte maturation, provides a physiological in situ model for investigating the role of c-raf-1 in proliferation and differentiation of chondrocytes. In this study, Northern blotting was first performed to examine the expression of mRNA for c-raf-1 in the embryonic chick tibial growth plate. It revealed that the normal levels of c-raf-1 mRNA were associated with the whole growth plate. We then investigated the localization of c-raf-1 mRNA and c-raf-1 protein in the growth plate by in situ hybridization and immunohistochemistry in order to determine whether c-raf-1 is involved in chondrocyte maturation. Our results showed that c-raf-1 mRNA and c-raf-1 protein were detected only in the hypertrophic zone. The data suggest involvement of this proto-oncogene in chondrocyte differentiation and/or hypertrophy rather than in proliferation.

Key words

Proto-oncogene C-raf-1 Growth plate Chondrocyte Differentiation 


Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.


  1. 1.
    Bishop JM (1983) Cellular oncogenes and retroviruses. Annu Rev Biochem 52:301–354Google Scholar
  2. 2.
    Der CJ, Cooper GM (1983) Altered gene products are associated with activation of cellular rats genes in human lung and colon carcinomas. Cell 32:201–208Google Scholar
  3. 3.
    Chang EH, Furth ME, Scolnick EM, Lowry DR (1982) Tumorigenesis of mammalian cells induced by a normal human gene homologous to the oncogene of Harvey murine sarcoma virus. Nature 297:479–484Google Scholar
  4. 4.
    Cooper GM (1982) Cellular transforming genes. Science 217: 801–806Google Scholar
  5. 5.
    Heidecker G, Huleihel M, Cleveland JL, Kolch W, Beck TW, Lloyd P, Pawson T, Rapp UR (1990) Mutational activation of c-raf-1 and definition of the minimal transforming sequence. Mol Cell Biol 10:2503–2512Google Scholar
  6. 6.
    Studzinski GP (1989) Oncogenes, growth, and the cell cycle: an overview. Cell Tissue Kinet 22:405–424Google Scholar
  7. 7.
    Littlewood TD, Evan GI (1990) The role of myc oncogenes in cell growth and differentiation. Adv Rent Res 4:67–79Google Scholar
  8. 8.
    Marshall GJ (1986) Oncogenes. J Cell Sci (suppl) 4:417–430Google Scholar
  9. 9.
    Bonner TI, Oppermann H, Seeburg P, Kerby SB, Gunnell MA, Young AC, Rapp UR (1986) The complete coding sequence of the human raf oncogene and the corresponding structure of the c-raf-1 gene. Nucleic Acids Res 14:1009–1015Google Scholar
  10. 10.
    Rapp UR, Heidecker G, Huleihel M, Cleveland JL, Choi WC, Pawson T, Ihle JN, Anderson WB (1988) Raf family serine threonine protein kinases in mitogen signal transduction. Cold Spring Harb Symp Quant Biol 53:173–184Google Scholar
  11. 11.
    Morrison DK, Kaplan DR, Rapp U, Roberts TM (1988) Signal transduction from membrane to cytoplasm: growth factors and membrane-bound oncogene products increase Raf-1 phosphorylation and associated protein kinase activity. Proc Natl Acad Sci USA 85:8855–8859Google Scholar
  12. 12.
    Olah Z, Komoly S, Nagashima N, Joo F, Rapp JR, Anderson WB (1991) Cerebral ischemia induces transient intracellular redistribution and intranuclear translocation of the raf protooncogene product in hippocampal pyramidal cells. Exp Brain Res 84:403–410Google Scholar
  13. 13.
    Baccarini M, Sabatini DM, App H, Rapp UR, Stanley ER (1990) Colony-stimulating factor-1 (CSF-1) stimulates temperature-dependent phosphorylation and activation of the RAF-1 protooncogene product. EMBO J 9:3649–3657Google Scholar
  14. 14.
    Blackshear PJ, Haupt DM, App H, Rapp UR (1990) Insulin activates the Raf-1 protein kinase. J Biol Chem 265:12131–12134Google Scholar
  15. 15.
    Kovacina KS, Yonezawa K, Brautigan DL, Tonks NK, Rapp UR, Roth RA (1990) Insulin activates the kinase activity of the Raf-1 proto-oncogene by increasing its serine phosphorylation. J Biol Chem 265:12115–12118Google Scholar
  16. 16.
    Turner B, Rapp U, App H, Greene M, Dobashi K, Reed J (1991) Interleukin 2 induces tyrosine phosphorylation and activation of p72–74 Raf-1 kinase in a T cell line. Proc Natl Acad Sci USA 88:1227–1231Google Scholar
  17. 17.
    Carroll MP, Clark Lewis I, Rapp UR, May WS (1990) Interleukin-3 and granulocyte macrophage colony-stimulating factor mediate rapid phosphorylation and activation of cytosolic c-raf. J Biol Chem 265:19812–19817Google Scholar
  18. 18.
    Kember NF, Walker KVR (1971) Control of bone growth in rats. Nature 229:428–429Google Scholar
  19. 19.
    Hunziker EB, Schenk RK (1989) Physiological mechanisms adopted by chondrocytes in regulating longitudinal bone growth in rats. J Physiol 414:55–71Google Scholar
  20. 20.
    Shirasawa H, Tomita Y, Sekiya S, Takamizawa H, Simizu B (1987) Intergration and transcription of human papillomavirus type 16 and 18 sequences in cell lines derived from cervical carcinomas. J Gen Virol 68:583–591Google Scholar
  21. 21.
    Tokunaga K, Taniguchi H, Yoda K, Shimizu M, Sakiyama S (1986) Nucleotide sequence of a full length cDNA for mouse cytoskeletal β-actin mRNA. Nucleic Acid Res 14:2829–2830Google Scholar
  22. 22.
    Hoefler H, Childers H, Montminy MR, Lechan RM, Goodman RH, Wolfe HJ (1986) In situ hybridization of methods for the detection of somatostatin mRNA in tissue sections using antisense RNA probes. Histochem J 18:597–604Google Scholar
  23. 23.
    Becker J, Schuppan D, Benzian H, Bals TH, Hahn EG, Cantaluppi CH, Reichart P (1986) Immunohistochemical distribution of collagens type IV, V and VI and of pro-collagens type I and III in human alveolar bone and dentin. J Histochem Cytochem 34:1417–1429Google Scholar
  24. 24.
    Morrison DK (1990) The Raf-1 kinase as a transducer of mitogenic signals. Cancer Cells 2:377–382Google Scholar
  25. 25.
    Heidecker G, Huleihel M, Cleveland IL, Kolch W, Beck TW, Lloyd P, Pawson T, Rapp UR (1990) Mutational activation of c-raf-1 and definition of the minimal transforming sequence. Mol Cell Biol 10:2503–2512Google Scholar
  26. 26.
    Nishida Y, Hata M, Ayaki T, Ryo H, Yamagata M, Shimizu K, Nishizuka Y (1988) Proliferation of both somatic and germ cells is affected in the Drosophila mutants of raf proto-oncogene. EMBO J 7:775–781Google Scholar
  27. 27.
    App H, Hazan R, Zilberstein A, Ullrich A, Schlessinger J, Rapp UR (1991) Epidermal growth factor (EGF) stimulates association and kinase activity of Raf-1 with the EGF receptor. Mol Cell Biol 11:913–919Google Scholar
  28. 28.
    Siegel JN, Klausner RD, Rapp UP, Samelson LE (1990) T cell antigen receptor engagement stimulates c-raf phosphorylation and induces c-raf associated kinase activity via a protein kinase C dependent pathway. J Biol Chem 265:18472–18480Google Scholar
  29. 29.
    Zmuidzinas A, Gould GW, Yager JD (1989) Expression of c-raf-1 and A-raf-1 during differentiation of 3T3-L1 preadipocyte fibroblasts into adipocytes. Biochem Biophys Res Commun 162:1180–1187Google Scholar
  30. 30.
    Ramp U, Gerharz CD, Doehmer J, Oster O, Gabbert HE (1992) Increase in proto oncogene raf expression precedes differentiation induction in different clonal rhabdomyosarcoma subpopulations. Anticancer Res 12:537–546Google Scholar

Copyright information

© Springer-Verlag New York Inc. 1994

Authors and Affiliations

  • Y. Kaneko
    • 1
  • H. Tanzawa
    • 1
  • K. Sato
    • 1
  1. 1.Department of Oral Surgery, School of MedicineChiba UniversityChibaJapan

Personalised recommendations