Abstract
The protein product of the v-myb oncogene of avian myeloblastosis virus, v-Myb, differs from its normal cellular counterpart, c-Myb, by (i) expression under the control of a strong viral long terminal repeat, (ii) truncation of both its amino and carboxyl termini, (iii) replacement of these termini by virally encoded residues, and (iv) substitution of 11 amino acid residues. We had previously shown that neither the virally encoded termini nor the amino acid substitutions are required for transformation by v-Myb. We have now constructed avian retroviruses that express full-length or singly truncated forms of c-Myb and have tested them for the transformation of chicken bone marrow cells. We conclude that truncation of either the amino or carboxyl terminus of c-Myb is sufficient for transformation. In contrast, the overexpression of full-length c-Myb does not result in transformation. We have also shown that the amino acid substitutions of v-Myb by themselves are not sufficient for the activation of c-Myb. Rather, the presence of either the normal amino or carboxyl terminus of c-Myb can suppress transformation when fused to v-Myb. Cells transformed by c-Myb proteins truncated at either their amino or carboxyl terminus appear to be granulated promyelocytes that express the Mim-1 protein. Cells transformed by a doubly truncated c-Myb protein are not granulated but do express the Mim-1 protein, in contrast to monoblasts transformed by v-Myb that neither contain granules nor express Mim-1. These results suggest that various alterations of c-Myb itself may determine the lineage of differentiating hematopoietic cells.
Full text
PDF









Images in this article
Selected References
These references are in PubMed. This may not be the complete list of references from this article.
- Beug H., Blundell P. A., Graf T. Reversibility of differentiation and proliferative capacity in avian myelomonocytic cells transformed by tsE26 leukemia virus. Genes Dev. 1987 May;1(3):277–286. doi: 10.1101/gad.1.3.277. [DOI] [PubMed] [Google Scholar]
- Biedenkapp H., Borgmeyer U., Sippel A. E., Klempnauer K. H. Viral myb oncogene encodes a sequence-specific DNA-binding activity. Nature. 1988 Oct 27;335(6193):835–837. doi: 10.1038/335835a0. [DOI] [PubMed] [Google Scholar]
- Bishop J. M. The molecular genetics of cancer. Science. 1987 Jan 16;235(4786):305–311. doi: 10.1126/science.3541204. [DOI] [PubMed] [Google Scholar]
- Boyle W. J., Lipsick J. S., Baluda M. A. Antibodies to the evolutionarily conserved amino-terminal region of the v-myb-encoded protein detect the c-myb protein in widely divergent metazoan species. Proc Natl Acad Sci U S A. 1986 Jul;83(13):4685–4689. doi: 10.1073/pnas.83.13.4685. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Chen C., Okayama H. High-efficiency transformation of mammalian cells by plasmid DNA. Mol Cell Biol. 1987 Aug;7(8):2745–2752. doi: 10.1128/mcb.7.8.2745. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Chen E. Y., Seeburg P. H. Supercoil sequencing: a fast and simple method for sequencing plasmid DNA. DNA. 1985 Apr;4(2):165–170. doi: 10.1089/dna.1985.4.165. [DOI] [PubMed] [Google Scholar]
- Clarke M. F., Kukowska-Latallo J. F., Westin E., Smith M., Prochownik E. V. Constitutive expression of a c-myb cDNA blocks Friend murine erythroleukemia cell differentiation. Mol Cell Biol. 1988 Feb;8(2):884–892. doi: 10.1128/mcb.8.2.884. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Clurman B. E., Hayward W. S. Multiple proto-oncogene activations in avian leukosis virus-induced lymphomas: evidence for stage-specific events. Mol Cell Biol. 1989 Jun;9(6):2657–2664. doi: 10.1128/mcb.9.6.2657. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Dudek H., Reddy E. P. Identification of two translational products for c-myb. Oncogene. 1989 Sep;4(9):1061–1066. [PubMed] [Google Scholar]
- Durban E. M., Boettiger D. Replicating, differentiated macrophages can serve as in vitro targets for transformation by avian myeloblastosis virus. J Virol. 1981 Jan;37(1):488–492. doi: 10.1128/jvi.37.1.488-492.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Evan G. I., Lewis G. K., Bishop J. M. Isolation of monoclonal antibodies specific for products of avian oncogene myb. Mol Cell Biol. 1984 Dec;4(12):2843–2850. doi: 10.1128/mcb.4.12.2843. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Garcia A., LaMontagne K., Reavis D., Stober-Grässer U., Lipsick J. S. Determinants of sequence-specific DNA-binding by p48v-myb. Oncogene. 1991 Feb;6(2):265–273. [PubMed] [Google Scholar]
- Gerondakis S., Bishop J. M. Structure of the protein encoded by the chicken proto-oncogene c-myb. Mol Cell Biol. 1986 Nov;6(11):3677–3684. doi: 10.1128/mcb.6.11.3677. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Golay J., Introna M., Graf T. A single point mutation in the v-ets oncogene affects both erythroid and myelomonocytic cell differentiation. Cell. 1988 Dec 23;55(6):1147–1158. doi: 10.1016/0092-8674(88)90259-0. [DOI] [PubMed] [Google Scholar]
- Gonda T. J., Buckmaster C., Ramsay R. G. Activation of c-myb by carboxy-terminal truncation: relationship to transformation of murine haemopoietic cells in vitro. EMBO J. 1989 Jun;8(6):1777–1783. doi: 10.1002/j.1460-2075.1989.tb03571.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Gonda T. J., Ramsay R. G., Johnson G. R. Murine myeloid cell lines derived by in vitro infection with recombinant c-myb retroviruses express myb from rearranged vector proviruses. EMBO J. 1989 Jun;8(6):1767–1775. doi: 10.1002/j.1460-2075.1989.tb03570.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Graf T. Two types of target cells for transformation with avian myelocytomatosis virus. Virology. 1973 Aug;54(2):398–413. doi: 10.1016/0042-6822(73)90152-9. [DOI] [PubMed] [Google Scholar]
- Graf T., von Kirchbach A., Beug H. Characterization of the hematopoietic target cells of AEV, MC29 and AMV avian leukemia viruses. Exp Cell Res. 1981 Feb;131(2):331–343. doi: 10.1016/0014-4827(81)90236-6. [DOI] [PubMed] [Google Scholar]
- Hughes S. H., Greenhouse J. J., Petropoulos C. J., Sutrave P. Adaptor plasmids simplify the insertion of foreign DNA into helper-independent retroviral vectors. J Virol. 1987 Oct;61(10):3004–3012. doi: 10.1128/jvi.61.10.3004-3012.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Hughes S., Mellstrom K., Kosik E., Tamanoi F., Brugge J. Mutation of a termination codon affects src initiation. Mol Cell Biol. 1984 Sep;4(9):1738–1746. doi: 10.1128/mcb.4.9.1738. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Ibanez C. E., Lipsick J. S. Structural and functional domains of the myb oncogene: requirements for nuclear transport, myeloid transformation, and colony formation. J Virol. 1988 Jun;62(6):1981–1988. doi: 10.1128/jvi.62.6.1981-1988.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Ibanez C. E., Lipsick J. S. trans activation of gene expression by v-myb. Mol Cell Biol. 1990 May;10(5):2285–2293. doi: 10.1128/mcb.10.5.2285. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Introna M., Golay J., Frampton J., Nakano T., Ness S. A., Graf T. Mutations in v-myb alter the differentiation of myelomonocytic cells transformed by the oncogene. Cell. 1990 Dec 21;63(6):1289–1297. doi: 10.1016/0092-8674(90)90424-d. [DOI] [PubMed] [Google Scholar]
- Kalkbrenner F., Guehmann S., Moelling K. Transcriptional activation by human c-myb and v-myb genes. Oncogene. 1990 May;5(5):657–661. [PubMed] [Google Scholar]
- Kanter M. R., Smith R. E., Hayward W. S. Rapid induction of B-cell lymphomas: insertional activation of c-myb by avian leukosis virus. J Virol. 1988 Apr;62(4):1423–1432. doi: 10.1128/jvi.62.4.1423-1432.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Katzen A. L., Kornberg T. B., Bishop J. M. Isolation of the proto-oncogene c-myb from D. melanogaster. Cell. 1985 Jun;41(2):449–456. doi: 10.1016/s0092-8674(85)80018-0. [DOI] [PubMed] [Google Scholar]
- Klempnauer K. H., Arnold H., Biedenkapp H. Activation of transcription by v-myb: evidence for two different mechanisms. Genes Dev. 1989 Oct;3(10):1582–1589. doi: 10.1101/gad.3.10.1582. [DOI] [PubMed] [Google Scholar]
- Kornfeld S., Beug H., Doederlein G., Graf T. Detection of avian hematopoietic cell surface antigens with monoclonal antibodies to myeloid cells. Their distribution on normal and leukemic cells of various lineages. Exp Cell Res. 1983 Feb;143(2):383–394. doi: 10.1016/0014-4827(83)90065-4. [DOI] [PubMed] [Google Scholar]
- Lane T., Ibanez C., Garcia A., Graf T., Lipsick J. Transformation by v-myb correlates with trans-activation of gene expression. Mol Cell Biol. 1990 Jun;10(6):2591–2598. doi: 10.1128/mcb.10.6.2591. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Leprince D., Gegonne A., Coll J., de Taisne C., Schneeberger A., Lagrou C., Stehelin D. A putative second cell-derived oncogene of the avian leukaemia retrovirus E26. Nature. 1983 Nov 24;306(5941):395–397. doi: 10.1038/306395a0. [DOI] [PubMed] [Google Scholar]
- Leutz A., Beug H., Graf T. Purification and characterization of cMGF, a novel chicken myelomonocytic growth factor. EMBO J. 1984 Dec 20;3(13):3191–3197. doi: 10.1002/j.1460-2075.1984.tb02278.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Lipsick J. S., Ibanez C. E., Baluda M. A. Expression of molecular clones of v-myb in avian and mammalian cells independently of transformation. J Virol. 1986 Aug;59(2):267–275. doi: 10.1128/jvi.59.2.267-275.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Lipsick J. S., Ibanez C. E. env-encoded residues are not required for transformation by p48v-myb. J Virol. 1987 Mar;61(3):933–936. doi: 10.1128/jvi.61.3.933-936.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Lipsick J. S. v-myb does not prevent the expression of c-myb in avian erythroblasts. J Virol. 1987 Oct;61(10):3284–3287. doi: 10.1128/jvi.61.10.3284-3287.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Lüscher B., Christenson E., Litchfield D. W., Krebs E. G., Eisenman R. N. Myb DNA binding inhibited by phosphorylation at a site deleted during oncogenic activation. Nature. 1990 Apr 5;344(6266):517–522. doi: 10.1038/344517a0. [DOI] [PubMed] [Google Scholar]
- McMahon J., Howe K. M., Watson R. J. The induction of Friend erythroleukaemia differentiation is markedly affected by expression of a transfected c-myb cDNA. Oncogene. 1988 Dec;3(6):717–720. [PubMed] [Google Scholar]
- Metz T., Graf T. v-myb and v-ets transform chicken erythroid cells and cooperate both in trans and in cis to induce distinct differentiation phenotypes. Genes Dev. 1991 Mar;5(3):369–380. doi: 10.1101/gad.5.3.369. [DOI] [PubMed] [Google Scholar]
- Mándi Y., Veromaa T., Baranji K., Miczák A., Béládi I., Toivanen P. Granulocyte-specific monoclonal antibody inhibiting cytotoxicity reactions in the chicken. Immunobiology. 1987 May;174(3):292–299. doi: 10.1016/s0171-2985(87)80004-9. [DOI] [PubMed] [Google Scholar]
- Ness S. A., Beug H., Graf T. v-myb dominance over v-myc in doubly transformed chick myelomonocytic cells. Cell. 1987 Oct 9;51(1):41–50. doi: 10.1016/0092-8674(87)90008-0. [DOI] [PubMed] [Google Scholar]
- Ness S. A., Marknell A., Graf T. The v-myb oncogene product binds to and activates the promyelocyte-specific mim-1 gene. Cell. 1989 Dec 22;59(6):1115–1125. doi: 10.1016/0092-8674(89)90767-8. [DOI] [PubMed] [Google Scholar]
- Nunn M. F., Seeburg P. H., Moscovici C., Duesberg P. H. Tripartite structure of the avian erythroblastosis virus E26 transforming gene. Nature. 1983 Nov 24;306(5941):391–395. doi: 10.1038/306391a0. [DOI] [PubMed] [Google Scholar]
- Peters C. W., Sippel A. E., Vingron M., Klempnauer K. H. Drosophila and vertebrate myb proteins share two conserved regions, one of which functions as a DNA-binding domain. EMBO J. 1987 Oct;6(10):3085–3090. doi: 10.1002/j.1460-2075.1987.tb02616.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Pizer E., Humphries E. H. RAV-1 insertional mutagenesis: disruption of the c-myb locus and development of avian B-cell lymphomas. J Virol. 1989 Apr;63(4):1630–1640. doi: 10.1128/jvi.63.4.1630-1640.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Ptashne M. How eukaryotic transcriptional activators work. Nature. 1988 Oct 20;335(6192):683–689. doi: 10.1038/335683a0. [DOI] [PubMed] [Google Scholar]
- Radke K., Beug H., Kornfeld S., Graf T. Transformation of both erythroid and myeloid cells by E26, an avian leukemia virus that contains the myb gene. Cell. 1982 Dec;31(3 Pt 2):643–653. doi: 10.1016/0092-8674(82)90320-8. [DOI] [PubMed] [Google Scholar]
- Ramsay R. G., Ishii S., Nishina Y., Soe G., Gonda T. J. Characterization of alternate and truncated forms of murine c-myb proteins. Oncogene Res. 1989;4(4):259–269. [PubMed] [Google Scholar]
- Rosson D. Effects of 5' and 3' truncations of the myb gene on the transforming ability of avian myeloblastosis virus (AMV). Virology. 1990 Apr;175(2):562–567. doi: 10.1016/0042-6822(90)90441-s. [DOI] [PubMed] [Google Scholar]
- Rosson D., Reddy E. P. Nucleotide sequence of chicken c-myb complementary DNA and implications for myb oncogene activation. Nature. 1986 Feb 13;319(6054):604–606. doi: 10.1038/319604a0. [DOI] [PubMed] [Google Scholar]
- Sakura H., Kanei-Ishii C., Nagase T., Nakagoshi H., Gonda T. J., Ishii S. Delineation of three functional domains of the transcriptional activator encoded by the c-myb protooncogene. Proc Natl Acad Sci U S A. 1989 Aug;86(15):5758–5762. doi: 10.1073/pnas.86.15.5758. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Shen-Ong G. L., Lüscher B., Eisenman R. N. A second c-myb protein is translated from an alternatively spliced mRNA expressed from normal and 5'-disrupted myb loci. Mol Cell Biol. 1989 Dec;9(12):5456–5463. doi: 10.1128/mcb.9.12.5456. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Shen-Ong G. L. The myb oncogene. Biochim Biophys Acta. 1990 Jun 1;1032(1):39–52. doi: 10.1016/0304-419x(90)90011-o. [DOI] [PubMed] [Google Scholar]
- Souza L. M., Strommer J. N., Hillyard R. L., Komaromy M. C., Baluda M. A. Cellular sequences are present in the presumptive avian myeloblastosis virus genome. Proc Natl Acad Sci U S A. 1980 Sep;77(9):5177–5181. doi: 10.1073/pnas.77.9.5177. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Stober-Grässer U., Lipsick J. S. Specific amino acid substitutions are not required for transformation by v-myb of avian myeloblastosis virus. J Virol. 1988 Mar;62(3):1093–1096. doi: 10.1128/jvi.62.3.1093-1096.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Weston K., Bishop J. M. Transcriptional activation by the v-myb oncogene and its cellular progenitor, c-myb. Cell. 1989 Jul 14;58(1):85–93. doi: 10.1016/0092-8674(89)90405-4. [DOI] [PubMed] [Google Scholar]