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. 1987 Jun 25;15(12):4809–4820. doi: 10.1093/nar/15.12.4809

Activation of human c-raf-1 by replacing the N-terminal region with different sequences.

T Tahira, M Ochiai, K Hayashi, M Nagao, T Sugimura
PMCID: PMC305919  PMID: 2955285

Abstract

Two transformants of NIH 3T3 cells, obtained by the transfection of human colon cancer and normal colon DNAs, contained activated c-raf-1. In both the activated c-raf-1, the 5' half of the c-raf-1 sequence was replaced by sequences other than c-raf-1 as a result of recombinations which occurred at the intron between exons 7 and 8. It was suggested, however, that these recombinations, which conferred the transforming activity on the c-raf-1, occurred during the transfection. In one case analyzed, characteristic sequences were found near the breakpoint and these may be involved in the recombination. It was found, upon analysing the structure of the cDNA derived from one of the activated c-raf-1, that fused mRNA had been transcribed from the recombined gene comprising the non-raf gene and c-raf-1. The mRNA possibly encodes a fused protein. One cDNA clone was derived from alternatively spliced mRNA, although its physiological role is unclear. On comparing the structure of the two human activated c-raf-1 and the rat activated c-raf which we have reported previously, it was revealed that, in these three cases, the sequences joined to the truncated c-raf(-1)1 were different. It was suggested from data which we and others have previously reported that various sequences could be capable of activating c-raf(-1) by replacing its 5' half.

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Selected References

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  1. Birchmeier C., Birnbaum D., Waitches G., Fasano O., Wigler M. Characterization of an activated human ros gene. Mol Cell Biol. 1986 Sep;6(9):3109–3116. doi: 10.1128/mcb.6.9.3109. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Bonner T. I., Oppermann H., Seeburg P., Kerby S. B., Gunnell M. A., Young A. C., Rapp U. R. The complete coding sequence of the human raf oncogene and the corresponding structure of the c-raf-1 gene. Nucleic Acids Res. 1986 Jan 24;14(2):1009–1015. doi: 10.1093/nar/14.2.1009. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Fritsch E. F., Lawn R. M., Maniatis T. Molecular cloning and characterization of the human beta-like globin gene cluster. Cell. 1980 Apr;19(4):959–972. doi: 10.1016/0092-8674(80)90087-2. [DOI] [PubMed] [Google Scholar]
  4. 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]
  5. 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]
  6. Ishikawa F., Takaku F., Hayashi K., Nagao M., Sugimura T. Activation of rat c-raf during transfection of hepatocellular carcinoma DNA. Proc Natl Acad Sci U S A. 1986 May;83(10):3209–3212. doi: 10.1073/pnas.83.10.3209. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Ishikawa F., Takaku F., Nagao M., Sugimura T. Cysteine-rich regions conserved in amino-terminal halves of raf gene family products and protein kinase C. Jpn J Cancer Res. 1986 Dec;77(12):1183–1187. [PubMed] [Google Scholar]
  8. Ishikawa F., Takaku F., Nagao M., Sugimura T. Rat c-raf oncogene activation by a rearrangement that produces a fused protein. Mol Cell Biol. 1987 Mar;7(3):1226–1232. doi: 10.1128/mcb.7.3.1226. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. 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]
  10. Jelinek W. R., Toomey T. P., Leinwand L., Duncan C. H., Biro P. A., Choudary P. V., Weissman S. M., Rubin C. M., Houck C. M., Deininger P. L. Ubiquitous, interspersed repeated sequences in mammalian genomes. Proc Natl Acad Sci U S A. 1980 Mar;77(3):1398–1402. doi: 10.1073/pnas.77.3.1398. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Kanehisa M., Klein P., Greif P., DeLisi C. Computer analysis and structure prediction of nucleic acids and proteins. Nucleic Acids Res. 1984 Jan 11;12(1 Pt 1):417–428. doi: 10.1093/nar/12.1part1.417. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Kyte J., Doolittle R. F. A simple method for displaying the hydropathic character of a protein. J Mol Biol. 1982 May 5;157(1):105–132. doi: 10.1016/0022-2836(82)90515-0. [DOI] [PubMed] [Google Scholar]
  13. Lathe R. Synthetic oligonucleotide probes deduced from amino acid sequence data. Theoretical and practical considerations. J Mol Biol. 1985 May 5;183(1):1–12. doi: 10.1016/0022-2836(85)90276-1. [DOI] [PubMed] [Google Scholar]
  14. 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]
  15. Mizusawa S., Nishimura S., Seela F. Improvement of the dideoxy chain termination method of DNA sequencing by use of deoxy-7-deazaguanosine triphosphate in place of dGTP. Nucleic Acids Res. 1986 Feb 11;14(3):1319–1324. doi: 10.1093/nar/14.3.1319. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Moelling K., Heimann B., Beimling P., Rapp U. R., Sander T. Serine- and threonine-specific protein kinase activities of purified gag-mil and gag-raf proteins. Nature. 1984 Dec 6;312(5994):558–561. doi: 10.1038/312558a0. [DOI] [PubMed] [Google Scholar]
  17. Mount S. M. A catalogue of splice junction sequences. Nucleic Acids Res. 1982 Jan 22;10(2):459–472. doi: 10.1093/nar/10.2.459. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. 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]
  19. 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]
  20. 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]
  21. 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]
  22. 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]
  23. 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]
  24. 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]
  25. 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]
  26. Young D., Waitches G., Birchmeier C., Fasano O., Wigler M. Isolation and characterization of a new cellular oncogene encoding a protein with multiple potential transmembrane domains. Cell. 1986 Jun 6;45(5):711–719. doi: 10.1016/0092-8674(86)90785-3. [DOI] [PubMed] [Google Scholar]

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