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. 1986 May;83(10):3209–3212. doi: 10.1073/pnas.83.10.3209

Activation of rat c-raf during transfection of hepatocellular carcinoma DNA.

F Ishikawa, F Takaku, K Hayashi, M Nagao, T Sugimura
PMCID: PMC323482  PMID: 3010283

Abstract

Rat c-raf was found to be activated in a transformant obtained with DNA of a hepatocellular carcinoma induced by 2-amino-3-methylimidazo[4,5-f]quinoline. This activated c-raf was cloned in a cosmid vector and actively transforming clones were obtained. Comparison of the restriction maps of this activated c-raf and cloned normal rat c-raf revealed a recombination in the 5'-terminal region of the activated form of this gene. The recombined DNA was shown to be actively transcribed and possibly to form a fused mRNA with c-raf, which is slightly smaller than normal c-raf mRNA. Since this recombination was not detected in the original tumor by Southern blot analysis, it presumably occurred during transfection.

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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. Bonner T. I., Kerby S. B., Sutrave P., Gunnell M. A., Mark G., Rapp U. R. Structure and biological activity of human homologs of the raf/mil oncogene. Mol Cell Biol. 1985 Jun;5(6):1400–1407. doi: 10.1128/mcb.5.6.1400. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Davies J., Jimenez A. A new selective agent for eukaryotic cloning vectors. Am J Trop Med Hyg. 1980 Sep;29(5 Suppl):1089–1092. doi: 10.4269/ajtmh.1980.29.1089. [DOI] [PubMed] [Google Scholar]
  4. Felton J. S., Knize M. G., Wood C., Wuebbles B. J., Healy S. K., Stuermer D. H., Bjeldanes L. F., Kimble B. J., Hatch F. T. Isolation and characterization of new mutagens from fried ground beef. Carcinogenesis. 1984 Jan;5(1):95–102. doi: 10.1093/carcin/5.1.95. [DOI] [PubMed] [Google Scholar]
  5. Feramisco J. R., Smart J. E., Burridge K., Helfman D. M., Thomas G. P. Co-existence of vinculin and a vinculin-like protein of higher molecular weight in smooth muscle. J Biol Chem. 1982 Sep 25;257(18):11024–11031. [PubMed] [Google Scholar]
  6. Frischauf A. M., Lehrach H., Poustka A., Murray N. Lambda replacement vectors carrying polylinker sequences. J Mol Biol. 1983 Nov 15;170(4):827–842. doi: 10.1016/s0022-2836(83)80190-9. [DOI] [PubMed] [Google Scholar]
  7. Fukui M., Yamamoto T., Kawai S., Maruo K., Toyoshima K. Detection of a raf-related and two other transforming DNA sequences in human tumors maintained in nude mice. Proc Natl Acad Sci U S A. 1985 Sep;82(17):5954–5958. doi: 10.1073/pnas.82.17.5954. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Hanahan D., Meselson M. Plasmid screening at high colony density. Gene. 1980 Jun;10(1):63–67. doi: 10.1016/0378-1119(80)90144-4. [DOI] [PubMed] [Google Scholar]
  9. Hayatsu H., Matsui Y., Ohara Y., Oka T., Hayatsu T. Characterization of mutagenic fractions in beef extract and in cooked ground beef. Use of blue-cotton for efficient extraction. Gan. 1983 Aug;74(4):472–482. [PubMed] [Google Scholar]
  10. Ishikawa F., Takaku F., Nagao M., Ochiai M., Hayashi K., Takayama S., Sugimura T. Activated oncogenes in a rat hepatocellular carcinoma induced by 2-amino-3-methylimidazo[4,5-f]quinoline. Jpn J Cancer Res. 1985 Jun;76(6):425–428. [PubMed] [Google Scholar]
  11. Ishikawa F., Takaku F., Ochiai M., Hayashi K., Hirohashi S., Terada M., Takayama S., Nagao M., Sugimura T. Activated c-raf gene in a rat hepatocellular carcinoma induced by 2-amino-3-methylimidazo[4,5-f]quinoline. Biochem Biophys Res Commun. 1985 Oct 15;132(1):186–192. doi: 10.1016/0006-291x(85)91005-8. [DOI] [PubMed] [Google Scholar]
  12. Lau Y. F., Kan Y. W. Versatile cosmid vectors for the isolation, expression, and rescue of gene sequences: studies with the human alpha-globin gene cluster. Proc Natl Acad Sci U S A. 1983 Sep;80(17):5225–5229. doi: 10.1073/pnas.80.17.5225. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Mölders H., Defesche J., Müller D., Bonner T. I., Rapp U. R., Müller R. Integration of transfected LTR sequences into the c-raf proto-oncogene: activation by promoter insertion. EMBO J. 1985 Mar;4(3):693–698. doi: 10.1002/j.1460-2075.1985.tb03685.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Ohgaki H., Kusama K., Matsukura N., Morino K., Hasegawa H., Sato S., Takayama S., Sugimura T. Carcinogenicity in mice of a mutagenic compound, 2-amino-3-methylimidazo[4,5-f]quinoline, from broiled sardine, cooked beef and beef extract. Carcinogenesis. 1984 Jul;5(7):921–924. doi: 10.1093/carcin/5.7.921. [DOI] [PubMed] [Google Scholar]
  15. Pellicer A., Wigler M., Axel R., Silverstein S. The transfer and stable integration of the HSV thymidine kinase gene into mouse cells. Cell. 1978 May;14(1):133–141. doi: 10.1016/0092-8674(78)90308-2. [DOI] [PubMed] [Google Scholar]
  16. Rapp U. R., Goldsborough M. D., Mark G. E., Bonner T. I., Groffen J., Reynolds F. H., Jr, Stephenson J. R. Structure and biological activity of v-raf, a unique oncogene transduced by a retrovirus. Proc Natl Acad Sci U S A. 1983 Jul;80(14):4218–4222. doi: 10.1073/pnas.80.14.4218. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Rapp U. R., Reynolds F. H., Jr, Stephenson J. R. New mammalian transforming retrovirus: demonstration of a polyprotein gene product. J Virol. 1983 Mar;45(3):914–924. doi: 10.1128/jvi.45.3.914-924.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Reddy E. P., Reynolds R. K., Santos E., Barbacid M. A point mutation is responsible for the acquisition of transforming properties by the T24 human bladder carcinoma oncogene. Nature. 1982 Nov 11;300(5888):149–152. doi: 10.1038/300149a0. [DOI] [PubMed] [Google Scholar]
  19. Shimizu K., Nakatsu Y., Sekiguchi M., Hokamura K., Tanaka K., Terada M., Sugimura T. Molecular cloning of an activated human oncogene, homologous to v-raf, from primary stomach cancer. Proc Natl Acad Sci U S A. 1985 Sep;82(17):5641–5645. doi: 10.1073/pnas.82.17.5641. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Southern E. M. Detection of specific sequences among DNA fragments separated by gel electrophoresis. J Mol Biol. 1975 Nov 5;98(3):503–517. doi: 10.1016/s0022-2836(75)80083-0. [DOI] [PubMed] [Google Scholar]
  21. Sugimura T. Carcinogenicity of mutagenic heterocyclic amines formed during the cooking process. Mutat Res. 1985 Jun-Jul;150(1-2):33–41. doi: 10.1016/0027-5107(85)90098-3. [DOI] [PubMed] [Google Scholar]
  22. Sukumar S., Notario V., Martin-Zanca D., Barbacid M. Induction of mammary carcinomas in rats by nitroso-methylurea involves malignant activation of H-ras-1 locus by single point mutations. Nature. 1983 Dec 15;306(5944):658–661. doi: 10.1038/306658a0. [DOI] [PubMed] [Google Scholar]
  23. Tabin C. J., Bradley S. M., Bargmann C. I., Weinberg R. A., Papageorge A. G., Scolnick E. M., Dhar R., Lowy D. R., Chang E. H. Mechanism of activation of a human oncogene. Nature. 1982 Nov 11;300(5888):143–149. doi: 10.1038/300143a0. [DOI] [PubMed] [Google Scholar]
  24. Takahashi M., Ritz J., Cooper G. M. Activation of a novel human transforming gene, ret, by DNA rearrangement. Cell. 1985 Sep;42(2):581–588. doi: 10.1016/0092-8674(85)90115-1. [DOI] [PubMed] [Google Scholar]
  25. Takayama S., Nakatsuru Y., Masuda M., Ohgaki H., Sato S., Sugimura T. Demonstration of carcinogenicity in F344 rats of 2-amino-3-methyl-imidazo[4,5-f]quinoline from broiled sardine, fried beef and beef extract. Gan. 1984 Jun;75(6):467–470. [PubMed] [Google Scholar]
  26. Wigler M., Silverstein S., Lee L. S., Pellicer A., Cheng Y. c., Axel R. Transfer of purified herpes virus thymidine kinase gene to cultured mouse cells. Cell. 1977 May;11(1):223–232. doi: 10.1016/0092-8674(77)90333-6. [DOI] [PubMed] [Google Scholar]

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