Abstract
The transforming protein encoded by the v-rel oncogene of the highly oncogenic avian retrovirus reticuloendotheliosis virus strain T (Rev-T) is a 59,000-dalton protein, p59v-rel. The mechanism by which p59v-rel induces transformation of early lymphoid cells is unknown. As a step towards understanding the mechanism of v-rel-induced transformation, we sought to establish the subcellular site of action of p59v-rel. In this report, we show that p59v-rel contains sequences that are necessary for its efficient localization in the nucleus of infected chicken embryo fibroblasts. These v-rel sequences when added to the normally cytoplasmic protein, beta-galactosidase, directed that protein to the nucleus. A mutation in the v-rel nuclear-localizing sequence did not affect the transforming function, although it did alter the nuclear-localizing function. The addition of a supplemental nuclear-localizing sequence from simian virus 40 large T-antigen to v-rel resulted in the expression of a transforming rel protein which was located exclusively in the nucleus of transformed spleen cells, in contrast to wild-type p59v-rel, which was largely cytoplasmic in transformed spleen cells. Our results support the hypothesis that v-rel encodes a protein which can act either in the nucleus or in the cytoplasm to transform spleen cells.
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- Alitalo K., Ramsay G., Bishop J. M., Pfeifer S. O., Colby W. W., Levinson A. D. Identification of nuclear proteins encoded by viral and cellular myc oncogenes. Nature. 1983 Nov 17;306(5940):274–277. doi: 10.1038/306274a0. [DOI] [PubMed] [Google Scholar]
- Barkas A. E., Brodeur D., Stavnezer E. Polyproteins containing a domain encoded by the V-SKI oncogene are located in the nuclei of SKV-transformed cells. Virology. 1986 May;151(1):131–138. doi: 10.1016/0042-6822(86)90111-x. [DOI] [PubMed] [Google Scholar]
- Casadaban M. J., Chou J., Cohen S. N. In vitro gene fusions that join an enzymatically active beta-galactosidase segment to amino-terminal fragments of exogenous proteins: Escherichia coli plasmid vectors for the detection and cloning of translational initiation signals. J Bacteriol. 1980 Aug;143(2):971–980. doi: 10.1128/jb.143.2.971-980.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Chen H. R., Yeh L. S., Barker W. C. Similarity between oncogenic v-rel protein and regulatory fnr protein. Nucleic Acids Res. 1986 May 12;14(9):3977–3977. doi: 10.1093/nar/14.9.3977. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 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]
- Donner P., Greiser-Wilke I., Moelling K. Nuclear localization and DNA binding of the transforming gene product of avian myelocytomatosis virus. Nature. 1982 Mar 18;296(5854):262–269. doi: 10.1038/296262a0. [DOI] [PubMed] [Google Scholar]
- Emerman M., Temin H. M. Genes with promoters in retrovirus vectors can be independently suppressed by an epigenetic mechanism. Cell. 1984 Dec;39(3 Pt 2):449–467. [PubMed] [Google Scholar]
- Franklin R. B., Maldonado R. L., Bose H. R. Isolation and characterization of reticuloendotheliosis virus transformed bone marrow cells. Intervirology. 1974;3(5-6):342–352. doi: 10.1159/000149771. [DOI] [PubMed] [Google Scholar]
- Garson K., Kang C. Y. Identification of the v-rel protein in REV-T transformed chicken bone marrow cells and expression in Cos1 cells. Biochem Biophys Res Commun. 1986 Jan 29;134(2):716–722. doi: 10.1016/s0006-291x(86)80479-x. [DOI] [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]
- Hall M. N., Hereford L., Herskowitz I. Targeting of E. coli beta-galactosidase to the nucleus in yeast. Cell. 1984 Apr;36(4):1057–1065. doi: 10.1016/0092-8674(84)90055-2. [DOI] [PubMed] [Google Scholar]
- Hann S. R., Abrams H. D., Rohrschneider L. R., Eisenman R. N. Proteins encoded by v-myc and c-myc oncogenes: identification and localization in acute leukemia virus transformants and bursal lymphoma cell lines. Cell. 1983 Oct;34(3):789–798. doi: 10.1016/0092-8674(83)90535-4. [DOI] [PubMed] [Google Scholar]
- Herzog N. K., Bose H. R., Jr Expression of the oncogene of avian reticuloendotheliosis virus in Escherichia coli and identification of the transforming protein in reticuloendotheliosis virus T-transformed cells. Proc Natl Acad Sci U S A. 1986 Feb;83(3):812–816. doi: 10.1073/pnas.83.3.812. [DOI] [PMC free article] [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]
- Kalderon D., Roberts B. L., Richardson W. D., Smith A. E. A short amino acid sequence able to specify nuclear location. Cell. 1984 Dec;39(3 Pt 2):499–509. doi: 10.1016/0092-8674(84)90457-4. [DOI] [PubMed] [Google Scholar]
- Kawai S., Nishizawa M. New procedure for DNA transfection with polycation and dimethyl sulfoxide. Mol Cell Biol. 1984 Jun;4(6):1172–1174. doi: 10.1128/mcb.4.6.1172. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Klempnauer K. H., Symonds G., Evan G. I., Bishop J. M. Subcellular localization of proteins encoded by oncogenes of avian myeloblastosis virus and avian leukemia virus E26 and by chicken c-myb gene. Cell. 1984 Jun;37(2):537–547. doi: 10.1016/0092-8674(84)90384-2. [DOI] [PubMed] [Google Scholar]
- Krippl B., Andrisani O., Jones N., Westphal H., Rosenberg M., Ferguson B. Adenovirus type 12 E1A protein expressed in Escherichia coli is functional upon transfer by microinjection or protoplast fusion into mammalian cells. J Virol. 1986 Aug;59(2):420–427. doi: 10.1128/jvi.59.2.420-427.1986. [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]
- Lanford R. E., Kanda P., Kennedy R. C. Induction of nuclear transport with a synthetic peptide homologous to the SV40 T antigen transport signal. Cell. 1986 Aug 15;46(4):575–582. doi: 10.1016/0092-8674(86)90883-4. [DOI] [PubMed] [Google Scholar]
- McKeon F. D., Tuffanelli D. L., Fukuyama K., Kirschner M. W. Autoimmune response directed against conserved determinants of nuclear envelope proteins in a patient with linear scleroderma. Proc Natl Acad Sci U S A. 1983 Jul;80(14):4374–4378. doi: 10.1073/pnas.80.14.4374. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Miller C. K., Temin H. M. Insertion of several different DNAs in reticuloendotheliosis virus strain T suppresses transformation by reducing the amount of subgenomic mRNA. J Virol. 1986 Apr;58(1):75–80. doi: 10.1128/jvi.58.1.75-80.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Paine P. L., Feldherr C. M. Nucleocytoplasmic exchange of macromolecules. Exp Cell Res. 1972 Sep;74(1):81–98. doi: 10.1016/0014-4827(72)90483-1. [DOI] [PubMed] [Google Scholar]
- Paine P. L. Nucleocytoplasmic movement of fluorescent tracers microinjected into living salivary gland cells. J Cell Biol. 1975 Sep;66(3):652–657. doi: 10.1083/jcb.66.3.652. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Pellman D., Garber E. A., Cross F. R., Hanafusa H. An N-terminal peptide from p60src can direct myristylation and plasma membrane localization when fused to heterologous proteins. 1985 Mar 28-Apr 3Nature. 314(6009):374–377. doi: 10.1038/314374a0. [DOI] [PubMed] [Google Scholar]
- Rice N. R., Copeland T. D., Simek S., Oroszlan S., Gilden R. V. Detection and characterization of the protein encoded by the v-rel oncogene. Virology. 1986 Mar;149(2):217–229. doi: 10.1016/0042-6822(86)90123-6. [DOI] [PubMed] [Google Scholar]
- Richardson W. D., Roberts B. L., Smith A. E. Nuclear location signals in polyoma virus large-T. Cell. 1986 Jan 17;44(1):77–85. doi: 10.1016/0092-8674(86)90486-1. [DOI] [PubMed] [Google Scholar]
- Roberts B. L., Richardson W. D., Smith A. E. The effect of protein context on nuclear location signal function. Cell. 1987 Jul 31;50(3):465–475. doi: 10.1016/0092-8674(87)90500-9. [DOI] [PubMed] [Google Scholar]
- Simek S. L., Stephens R. M., Rice N. R. Localization of the v-rel protein in reticuloendotheliosis virus strain T-transformed lymphoid cells. J Virol. 1986 Jul;59(1):120–126. doi: 10.1128/jvi.59.1.120-126.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Stephens R. M., Rice N. R., Hiebsch R. R., Bose H. R., Jr, Gilden R. V. Nucleotide sequence of v-rel: the oncogene of reticuloendotheliosis virus. Proc Natl Acad Sci U S A. 1983 Oct;80(20):6229–6233. doi: 10.1073/pnas.80.20.6229. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Sylla B. S., Temin H. M. Activation of oncogenicity of the c-rel proto-oncogene. Mol Cell Biol. 1986 Dec;6(12):4709–4716. doi: 10.1128/mcb.6.12.4709. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Vogt P. K., Bos T. J., Doolittle R. F. Homology between the DNA-binding domain of the GCN4 regulatory protein of yeast and the carboxyl-terminal region of a protein coded for by the oncogene jun. Proc Natl Acad Sci U S A. 1987 May;84(10):3316–3319. doi: 10.1073/pnas.84.10.3316. [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]
- Watanabe S., Temin H. M. Encapsidation sequences for spleen necrosis virus, an avian retrovirus, are between the 5' long terminal repeat and the start of the gag gene. Proc Natl Acad Sci U S A. 1982 Oct;79(19):5986–5990. doi: 10.1073/pnas.79.19.5986. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Welsh J. D., Swimmer C., Cocke T., Shenk T. A second domain of simian virus 40 T antigen in which mutations can alter the cellular localization of the antigen. Mol Cell Biol. 1986 Jun;6(6):2207–2212. doi: 10.1128/mcb.6.6.2207. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Wilhelmsen K. C., Eggleton K., Temin H. M. Nucleic acid sequences of the oncogene v-rel in reticuloendotheliosis virus strain T and its cellular homolog, the proto-oncogene c-rel. J Virol. 1984 Oct;52(1):172–182. doi: 10.1128/jvi.52.1.172-182.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Young J. C., Martin G. S. Cellular localization of c-fps gene product NCP98. J Virol. 1984 Dec;52(3):913–918. doi: 10.1128/jvi.52.3.913-918.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]