Abstract
By making site-directed mutations in the avian retroviral oncogene v-rel, we created two temperature-sensitive (ts) transforming mutants; these changes were analogous to mutations previously shown to confer a ts function onto the Dorsal protein of Drosophila melanogaster. Chicken spleen cells infected with the ts v-rel mutants formed colonies in agar at 36.5 degrees C but not at 41.5 degrees C. In addition, spleen cells derived from the ts v-rel-transformed colonies could be propagated in liquid culture at 36.5 degrees C but rapidly senesced at 41.5 degrees C. Both mutant v-Rel proteins were also ts for DNA binding in vitro. These mutants may be valuable for identifying genes directly regulated by v-rel.
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- Baeuerle P. A. The inducible transcription activator NF-kappa B: regulation by distinct protein subunits. Biochim Biophys Acta. 1991 Apr 16;1072(1):63–80. doi: 10.1016/0304-419x(91)90007-8. [DOI] [PubMed] [Google Scholar]
- Ballard D. W., Walker W. H., Doerre S., Sista P., Molitor J. A., Dixon E. P., Peffer N. J., Hannink M., Greene W. C. The v-rel oncogene encodes a kappa B enhancer binding protein that inhibits NF-kappa B function. Cell. 1990 Nov 16;63(4):803–814. doi: 10.1016/0092-8674(90)90146-6. [DOI] [PubMed] [Google Scholar]
- Boehmelt G., Walker A., Kabrun N., Mellitzer G., Beug H., Zenke M., Enrietto P. J. Hormone-regulated v-rel estrogen receptor fusion protein: reversible induction of cell transformation and cellular gene expression. EMBO J. 1992 Dec;11(12):4641–4652. doi: 10.1002/j.1460-2075.1992.tb05566.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Bose H. R., Jr The Rel family: models for transcriptional regulation and oncogenic transformation. Biochim Biophys Acta. 1992 Sep 14;1114(1):1–17. doi: 10.1016/0304-419x(92)90002-g. [DOI] [PubMed] [Google Scholar]
- Capobianco A. J., Gilmore T. D. A conditional mutant of vRel containing sequences from the human estrogen receptor. Virology. 1993 Mar;193(1):160–170. doi: 10.1006/viro.1993.1112. [DOI] [PubMed] [Google Scholar]
- Capobianco A. J., Gilmore T. D. Repression of the chicken c-rel promoter by vRel in chicken embryo fibroblasts is not mediated through a consensus NF-kappa B binding site. Oncogene. 1991 Dec;6(12):2203–2210. [PubMed] [Google Scholar]
- Capobianco A. J., Simmons D. L., Gilmore T. D. Cloning and expression of a chicken c-rel cDNA: unlike p59v-rel, p68c-rel is a cytoplasmic protein in chicken embryo fibroblasts. Oncogene. 1990 Mar;5(3):257–265. [PubMed] [Google Scholar]
- Chen I. S., Temin H. M. Substitution of 5' helper virus sequences into non-rel portion of reticuloendotheliosis virus strain T suppresses transformation of chicken spleen cells. Cell. 1982 Nov;31(1):111–120. doi: 10.1016/0092-8674(82)90410-x. [DOI] [PubMed] [Google Scholar]
- Gilmore T. D. Malignant transformation by mutant Rel proteins. Trends Genet. 1991 Oct;7(10):318–322. doi: 10.1016/0168-9525(91)90421-l. [DOI] [PubMed] [Google Scholar]
- Gilmore T. D. NF-kappa B, KBF1, dorsal, and related matters. Cell. 1990 Sep 7;62(5):841–843. doi: 10.1016/0092-8674(90)90257-f. [DOI] [PubMed] [Google Scholar]
- Gilmore T. D. Role of rel family genes in normal and malignant lymphoid cell growth. Cancer Surv. 1992;15:69–87. [PubMed] [Google Scholar]
- Gilmore T. D., Temin H. M. Different localization of the product of the v-rel oncogene in chicken fibroblasts and spleen cells correlates with transformation by REV-T. Cell. 1986 Mar 14;44(5):791–800. doi: 10.1016/0092-8674(86)90845-7. [DOI] [PubMed] [Google Scholar]
- Gilmore T. D., Temin H. M. v-rel oncoproteins in the nucleus and in the cytoplasm transform chicken spleen cells. J Virol. 1988 Mar;62(3):703–714. doi: 10.1128/jvi.62.3.703-714.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Gélinas C., Temin H. M. The v-rel oncogene encodes a cell-specific transcriptional activator of certain promoters. Oncogene. 1988 Oct;3(4):349–355. [PubMed] [Google Scholar]
- Hirt B. Selective extraction of polyoma DNA from infected mouse cell cultures. J Mol Biol. 1967 Jun 14;26(2):365–369. doi: 10.1016/0022-2836(67)90307-5. [DOI] [PubMed] [Google Scholar]
- Hoelzer J. D., Lewis R. B., Wasmuth C. R., Bose H. R., Jr Hematopoietic cell transformation by reticuloendotheliosis virus: characterization of the genetic defect. Virology. 1980 Jan 30;100(2):462–474. doi: 10.1016/0042-6822(80)90536-x. [DOI] [PubMed] [Google Scholar]
- Isoda K., Roth S., Nüsslein-Volhard C. The functional domains of the Drosophila morphogen dorsal: evidence from the analysis of mutants. Genes Dev. 1992 Apr;6(4):619–630. doi: 10.1101/gad.6.4.619. [DOI] [PubMed] [Google Scholar]
- Kabrun N., Hodgson J. W., Doemer M., Mak G., Franza B. R., Jr, Enrietto P. J. Interaction of the v-rel protein with an NF-kappa B DNA binding site. Proc Natl Acad Sci U S A. 1991 Mar 1;88(5):1783–1787. doi: 10.1073/pnas.88.5.1783. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Kamens J., Richardson P., Mosialos G., Brent R., Gilmore T. Oncogenic transformation by vrel requires an amino-terminal activation domain. Mol Cell Biol. 1990 Jun;10(6):2840–2847. doi: 10.1128/mcb.10.6.2840. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Kochel T., Rice N. R. v-rel- and c-rel-protein complexes bind to the NF-kappa B site in vitro. Oncogene. 1992 Mar;7(3):567–572. [PubMed] [Google Scholar]
- Kumar S., Rabson A. B., Gélinas C. The RxxRxRxxC motif conserved in all Rel/kappa B proteins is essential for the DNA-binding activity and redox regulation of the v-Rel oncoprotein. Mol Cell Biol. 1992 Jul;12(7):3094–3106. doi: 10.1128/mcb.12.7.3094. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Kunkel T. A. Rapid and efficient site-specific mutagenesis without phenotypic selection. Proc Natl Acad Sci U S A. 1985 Jan;82(2):488–492. doi: 10.1073/pnas.82.2.488. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Matthews J. R., Kaszubska W., Turcatti G., Wells T. N., Hay R. T. Role of cysteine62 in DNA recognition by the P50 subunit of NF-kappa B. Nucleic Acids Res. 1993 Apr 25;21(8):1727–1734. doi: 10.1093/nar/21.8.1727. [DOI] [PMC free article] [PubMed] [Google Scholar]
- McDonnell P. C., Kumar S., Rabson A. B., Gélinas C. Transcriptional activity of rel family proteins. Oncogene. 1992 Jan;7(1):163–170. [PubMed] [Google Scholar]
- Mosialos G., Gilmore T. D. v-Rel and c-Rel are differentially affected by mutations at a consensus protein kinase recognition sequence. Oncogene. 1993 Mar;8(3):721–730. [PubMed] [Google Scholar]
- Mosialos G., Hamer P., Capobianco A. J., Laursen R. A., Gilmore T. D. A protein kinase-A recognition sequence is structurally linked to transformation by p59v-rel and cytoplasmic retention of p68c-rel. Mol Cell Biol. 1991 Dec;11(12):5867–5877. doi: 10.1128/mcb.11.12.5867. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Richardson P. M., Gilmore T. D. vRel is an inactive member of the Rel family of transcriptional activating proteins. J Virol. 1991 Jun;65(6):3122–3130. doi: 10.1128/jvi.65.6.3122-3130.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Sarkar S., Gilmore T. D. Transformation by the vRel oncoprotein requires sequences carboxy-terminal to the Rel homology domain. Oncogene. 1993 Aug;8(8):2245–2252. [PubMed] [Google Scholar]
- Sif S., Capobianco A. J., Gilmore T. D. The v-Rel oncoprotein increases expression from Sp1 site-containing promoters in chicken embryo fibroblasts. Oncogene. 1993 Sep;8(9):2501–2509. [PubMed] [Google Scholar]
- Walker W. H., Stein B., Ganchi P. A., Hoffman J. A., Kaufman P. A., Ballard D. W., Hannink M., Greene W. C. The v-rel oncogene: insights into the mechanism of transcriptional activation, repression, and transformation. J Virol. 1992 Aug;66(8):5018–5029. doi: 10.1128/jvi.66.8.5018-5029.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Watanabe S., Temin H. M. Construction of a helper cell line for avian reticuloendotheliosis virus cloning vectors. Mol Cell Biol. 1983 Dec;3(12):2241–2249. doi: 10.1128/mcb.3.12.2241. [DOI] [PMC free article] [PubMed] [Google Scholar]