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. 1988 May;8(5):2251–2256. doi: 10.1128/mcb.8.5.2251

Detection of fos protein during osteogenesis by monoclonal antibodies.

P De Togni 1, H Niman 1, V Raymond 1, P Sawchenko 1, I M Verma 1
PMCID: PMC363412  PMID: 3133556

Abstract

We have generated monoclonal antibodies by using a synthetic peptide corresponding to amino acid positions 4 to 17 of the human fos protein. The antibodies detected both v- and c-fos proteins by immunoprecipitation, immunoblotting, and indirect immunofluorescence. The monoclonal antibodies not only identified the fos protein complex with the cellular 39-kilodalton protein, but also recognized the modified forms of the mouse, rat, and human fos proteins. In day-17 rat embryos, nuclear-staining fos protein could be identified in the cartilage by immunohistochemical staining.

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Selected References

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  1. Barber J. R., Verma I. M. Modification of fos proteins: phosphorylation of c-fos, but not v-fos, is stimulated by 12-tetradecanoyl-phorbol-13-acetate and serum. Mol Cell Biol. 1987 Jun;7(6):2201–2211. doi: 10.1128/mcb.7.6.2201. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Chamberlain J. P. Fluorographic detection of radioactivity in polyacrylamide gels with the water-soluble fluor, sodium salicylate. Anal Biochem. 1979 Sep 15;98(1):132–135. doi: 10.1016/0003-2697(79)90716-4. [DOI] [PubMed] [Google Scholar]
  3. 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]
  4. Curran T., Miller A. D., Zokas L., Verma I. M. Viral and cellular fos proteins: a comparative analysis. Cell. 1984 Feb;36(2):259–268. doi: 10.1016/0092-8674(84)90219-8. [DOI] [PubMed] [Google Scholar]
  5. Curran T., Van Beveren C., Verma I. M. Viral and cellular fos proteins are complexed with a 39,000-dalton cellular protein. Mol Cell Biol. 1985 Jan;5(1):167–172. doi: 10.1128/mcb.5.1.167. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Distel R. J., Ro H. S., Rosen B. S., Groves D. L., Spiegelman B. M. Nucleoprotein complexes that regulate gene expression in adipocyte differentiation: direct participation of c-fos. Cell. 1987 Jun 19;49(6):835–844. doi: 10.1016/0092-8674(87)90621-0. [DOI] [PubMed] [Google Scholar]
  7. Dony C., Gruss P. Proto-oncogene c-fos expression in growth regions of fetal bone and mesodermal web tissue. Nature. 1987 Aug 20;328(6132):711–714. doi: 10.1038/328711a0. [DOI] [PubMed] [Google Scholar]
  8. Finkel M. P., Biskis B. O., Jinkins P. B. Virus induction of osteosarcomas in mice. Science. 1966 Feb 11;151(3711):698–701. doi: 10.1126/science.151.3711.698. [DOI] [PubMed] [Google Scholar]
  9. Fujiwara K. T., Ashida K., Nishina H., Iba H., Miyajima N., Nishizawa M., Kawai S. The chicken c-fos gene: cloning and nucleotide sequence analysis. J Virol. 1987 Dec;61(12):4012–4018. doi: 10.1128/jvi.61.12.4012-4018.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Glenney J. Antibody probing of western blots which have been stained with india ink. Anal Biochem. 1986 Aug 1;156(2):315–319. doi: 10.1016/0003-2697(86)90259-9. [DOI] [PubMed] [Google Scholar]
  11. Greenberg M. E., Ziff E. B. Stimulation of 3T3 cells induces transcription of the c-fos proto-oncogene. Nature. 1984 Oct 4;311(5985):433–438. doi: 10.1038/311433a0. [DOI] [PubMed] [Google Scholar]
  12. Holt J. T., Gopal T. V., Moulton A. D., Nienhuis A. W. Inducible production of c-fos antisense RNA inhibits 3T3 cell proliferation. Proc Natl Acad Sci U S A. 1986 Jul;83(13):4794–4798. doi: 10.1073/pnas.83.13.4794. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Houghten R. A. General method for the rapid solid-phase synthesis of large numbers of peptides: specificity of antigen-antibody interaction at the level of individual amino acids. Proc Natl Acad Sci U S A. 1985 Aug;82(15):5131–5135. doi: 10.1073/pnas.82.15.5131. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Kruijer W., Cooper J. A., Hunter T., Verma I. M. Platelet-derived growth factor induces rapid but transient expression of the c-fos gene and protein. Nature. 1984 Dec 20;312(5996):711–716. doi: 10.1038/312711a0. [DOI] [PubMed] [Google Scholar]
  15. 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]
  16. Lerner R. A., Green N., Alexander H., Liu F. T., Sutcliffe J. G., Shinnick T. M. Chemically synthesized peptides predicted from the nucleotide sequence of the hepatitis B virus genome elicit antibodies reactive with the native envelope protein of Dane particles. Proc Natl Acad Sci U S A. 1981 Jun;78(6):3403–3407. doi: 10.1073/pnas.78.6.3403. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Müller R. Cellular and viral fos genes: structure, regulation of expression and biological properties of their encoded products. Biochim Biophys Acta. 1986;823(3):207–225. doi: 10.1016/0304-419x(86)90003-x. [DOI] [PubMed] [Google Scholar]
  18. Müller R., Verma I. M. Expression of cellular oncogenes. Curr Top Microbiol Immunol. 1984;112:73–115. doi: 10.1007/978-3-642-69677-0_4. [DOI] [PubMed] [Google Scholar]
  19. Niman H. L., Elder J. H. Molecular dissection of Rauscher virus gp70 by using monoclonal antibodies: localization of acquired sequences of related envelope gene recombinants. Proc Natl Acad Sci U S A. 1980 Aug;77(8):4524–4528. doi: 10.1073/pnas.77.8.4524. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Niman H. L., Thompson A. M., Yu A., Markman M., Willems J. J., Herwig K. R., Habib N. A., Wood C. B., Houghten R. A., Lerner R. A. Anti-peptide antibodies detect oncogene-related proteins in urine. Proc Natl Acad Sci U S A. 1985 Dec;82(23):7924–7928. doi: 10.1073/pnas.82.23.7924. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Nishikura K., Murray J. M. Antisense RNA of proto-oncogene c-fos blocks renewed growth of quiescent 3T3 cells. Mol Cell Biol. 1987 Feb;7(2):639–649. doi: 10.1128/mcb.7.2.639. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Rüther U., Garber C., Komitowski D., Müller R., Wagner E. F. Deregulated c-fos expression interferes with normal bone development in transgenic mice. 1987 Jan 29-Feb 4Nature. 325(6103):412–416. doi: 10.1038/325412a0. [DOI] [PubMed] [Google Scholar]
  23. Van Beveren C., van Straaten F., Curran T., Müller R., Verma I. M. Analysis of FBJ-MuSV provirus and c-fos (mouse) gene reveals that viral and cellular fos gene products have different carboxy termini. Cell. 1983 Apr;32(4):1241–1255. doi: 10.1016/0092-8674(83)90306-9. [DOI] [PubMed] [Google Scholar]
  24. Verma I. M., Graham W. R. The fos oncogene. Adv Cancer Res. 1987;49:29–52. doi: 10.1016/s0065-230x(08)60793-9. [DOI] [PubMed] [Google Scholar]
  25. Verma I. M., Sassone-Corsi P. Proto-oncogene fos: complex but versatile regulation. Cell. 1987 Nov 20;51(4):513–514. doi: 10.1016/0092-8674(87)90115-2. [DOI] [PubMed] [Google Scholar]
  26. van Straaten F., Müller R., Curran T., Van Beveren C., Verma I. M. Complete nucleotide sequence of a human c-onc gene: deduced amino acid sequence of the human c-fos protein. Proc Natl Acad Sci U S A. 1983 Jun;80(11):3183–3187. doi: 10.1073/pnas.80.11.3183. [DOI] [PMC free article] [PubMed] [Google Scholar]

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