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. 1996 Nov;70(11):8089–8097. doi: 10.1128/jvi.70.11.8089-8097.1996

Transforming activity of retroviral genomes encoding Gag-Axl fusion proteins.

Q K Zhang 1, S Boast 1, K de los Santos 1, M Begemann 1, S P Goff 1
PMCID: PMC190883  PMID: 8892934

Abstract

Retroviral genomes encoding a portion of the Moloney murine leukemia virus Gag protein fused to portions of the murine axl cDNA were constructed so as to mimic naturally occurring transforming viruses. Virus MA1 retained 5 amino acids of the extracellular domain and the complete transmembrane and intracellular domains of Axl; virus MA2 retained only the intracellular Axl sequences beginning 33 amino acids downstream of the transmembrane region. Although both viruses could transform NIH 3T3 cells, they induced different morphological changes. MA1 transformants became elongated and assumed a cross-hatched pattern, while MA2 transformants were round and very refractile and grew to high density. Gag-Axl and Glyco-Gag-Axl proteins were detected in both types of transformed cells and were predominantly localized to the cytoplasmic compartment. When cell-free v-axl virus supernatants were introduced into wild-type BALB/c neonates, Rag-2-deficient mice, or c-myc transgenic mice, they did not cause tumors in a 3-month period. However, MA2-transformed NIH 3T3 cells, but not MA1 or control cells, could establish sarcomas by subcutaneous or intraperitoneal injection into BALB/c neonates. These results show that the transforming potential of the axl gene can be activated by truncation of the extracellular domain of the receptor and fusion of the remaining sequence to the gag gene.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Adams J. M., Harris A. W., Pinkert C. A., Corcoran L. M., Alexander W. S., Cory S., Palmiter R. D., Brinster R. L. The c-myc oncogene driven by immunoglobulin enhancers induces lymphoid malignancy in transgenic mice. Nature. 1985 Dec 12;318(6046):533–538. doi: 10.1038/318533a0. [DOI] [PubMed] [Google Scholar]
  2. Alexander W. S., Adams J. M., Cory S. Oncogene cooperation in lymphocyte transformation: malignant conversion of E mu-myc transgenic pre-B cells in vitro is enhanced by v-H-ras or v-raf but not v-abl. Mol Cell Biol. 1989 Jan;9(1):67–73. doi: 10.1128/mcb.9.1.67. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Alin K., Goff S. P. Mutational analysis of interactions between the Gag precursor proteins of murine leukemia viruses. Virology. 1996 Feb 15;216(2):418–424. doi: 10.1006/viro.1996.0078. [DOI] [PubMed] [Google Scholar]
  4. Anderson S. J., Gonda M. A., Rettenmier C. W., Sherr C. J. Subcellular localization of glycoproteins encoded by the viral oncogene v-fms. J Virol. 1984 Sep;51(3):730–741. doi: 10.1128/jvi.51.3.730-741.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Bassiri M., Privalsky M. L. Transmembrane domain of the AEV erb B oncogene protein is not required for partial manifestation of the transformed phenotype. Virology. 1987 Jul;159(1):20–30. doi: 10.1016/0042-6822(87)90343-6. [DOI] [PubMed] [Google Scholar]
  6. Bellacosa A., Testa J. R., Staal S. P., Tsichlis P. N. A retroviral oncogene, akt, encoding a serine-threonine kinase containing an SH2-like region. Science. 1991 Oct 11;254(5029):274–277. doi: 10.1126/science.254.5029.274. [DOI] [PubMed] [Google Scholar]
  7. Berlioz C., Darlix J. L. An internal ribosomal entry mechanism promotes translation of murine leukemia virus gag polyprotein precursors. J Virol. 1995 Apr;69(4):2214–2222. doi: 10.1128/jvi.69.4.2214-2222.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Berns A. Tumorigenesis in transgenic mice: identification and characterization of synergizing oncogenes. J Cell Biochem. 1991 Oct;47(2):130–135. doi: 10.1002/jcb.240470206. [DOI] [PubMed] [Google Scholar]
  9. Colicelli J., Goff S. P. Sequence and spacing requirements of a retrovirus integration site. J Mol Biol. 1988 Jan 5;199(1):47–59. doi: 10.1016/0022-2836(88)90378-6. [DOI] [PubMed] [Google Scholar]
  10. Coulier F., Martin-Zanca D., Ernst M., Barbacid M. Mechanism of activation of the human trk oncogene. Mol Cell Biol. 1989 Jan;9(1):15–23. doi: 10.1128/mcb.9.1.15. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Downward J., Yarden Y., Mayes E., Scrace G., Totty N., Stockwell P., Ullrich A., Schlessinger J., Waterfield M. D. Close similarity of epidermal growth factor receptor and v-erb-B oncogene protein sequences. Nature. 1984 Feb 9;307(5951):521–527. doi: 10.1038/307521a0. [DOI] [PubMed] [Google Scholar]
  12. Fantl W. J., Johnson D. E., Williams L. T. Signalling by receptor tyrosine kinases. Annu Rev Biochem. 1993;62:453–481. doi: 10.1146/annurev.bi.62.070193.002321. [DOI] [PubMed] [Google Scholar]
  13. Faust M., Ebensperger C., Schulz A. S., Schleithoff L., Hameister H., Bartram C. R., Janssen J. W. The murine ufo receptor: molecular cloning, chromosomal localization and in situ expression analysis. Oncogene. 1992 Jul;7(7):1287–1293. [PubMed] [Google Scholar]
  14. Fridell Y. W., Jin Y., Quilliam L. A., Burchert A., McCloskey P., Spizz G., Varnum B., Der C., Liu E. T. Differential activation of the Ras/extracellular-signal-regulated protein kinase pathway is responsible for the biological consequences induced by the Axl receptor tyrosine kinase. Mol Cell Biol. 1996 Jan;16(1):135–145. doi: 10.1128/mcb.16.1.135. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Golub T. R., Barker G. F., Lovett M., Gilliland D. G. Fusion of PDGF receptor beta to a novel ets-like gene, tel, in chronic myelomonocytic leukemia with t(5;12) chromosomal translocation. Cell. 1994 Apr 22;77(2):307–316. doi: 10.1016/0092-8674(94)90322-0. [DOI] [PubMed] [Google Scholar]
  16. Greco A., Pierotti M. A., Bongarzone I., Pagliardini S., Lanzi C., Della Porta G. TRK-T1 is a novel oncogene formed by the fusion of TPR and TRK genes in human papillary thyroid carcinomas. Oncogene. 1992 Feb;7(2):237–242. [PubMed] [Google Scholar]
  17. Harris A. W., Pinkert C. A., Crawford M., Langdon W. Y., Brinster R. L., Adams J. M. The E mu-myc transgenic mouse. A model for high-incidence spontaneous lymphoma and leukemia of early B cells. J Exp Med. 1988 Feb 1;167(2):353–371. doi: 10.1084/jem.167.2.353. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Janssen J. W., Schulz A. S., Steenvoorden A. C., Schmidberger M., Strehl S., Ambros P. F., Bartram C. R. A novel putative tyrosine kinase receptor with oncogenic potential. Oncogene. 1991 Nov;6(11):2113–2120. [PubMed] [Google Scholar]
  19. Jia R., Mayer B. J., Hanafusa T., Hanafusa H. A novel oncogene, v-ryk, encoding a truncated receptor tyrosine kinase is transduced into the RPL30 virus without loss of viral sequences. J Virol. 1992 Oct;66(10):5975–5987. doi: 10.1128/jvi.66.10.5975-5987.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Langdon W. Y., Harris A. W., Cory S. Acceleration of B-lymphoid tumorigenesis in E mu-myc transgenic mice by v-H-ras and v-raf but not v-abl. Oncogene Res. 1989;4(4):253–258. [PubMed] [Google Scholar]
  21. Lazo P. A., Tsichlis P. N. Biology and pathogenesis of retroviruses. Semin Oncol. 1990 Jun;17(3):269–294. [PubMed] [Google Scholar]
  22. Manger R., Najita L., Nichols E. J., Hakomori S., Rohrschneider L. Cell surface expression of the McDonough strain of feline sarcoma virus fms gene product (gp 140fms). Cell. 1984 Dec;39(2 Pt 1):327–337. doi: 10.1016/0092-8674(84)90011-4. [DOI] [PubMed] [Google Scholar]
  23. Mark G. E., Rapp U. R. Primary structure of v-raf: relatedness to the src family of oncogenes. Science. 1984 Apr 20;224(4646):285–289. doi: 10.1126/science.6324342. [DOI] [PubMed] [Google Scholar]
  24. McCutchan J. H., Pagano J. S. Enchancement of the infectivity of simian virus 40 deoxyribonucleic acid with diethylaminoethyl-dextran. J Natl Cancer Inst. 1968 Aug;41(2):351–357. [PubMed] [Google Scholar]
  25. McWhirter J. R., Galasso D. L., Wang J. Y. A coiled-coil oligomerization domain of Bcr is essential for the transforming function of Bcr-Abl oncoproteins. Mol Cell Biol. 1993 Dec;13(12):7587–7595. doi: 10.1128/mcb.13.12.7587. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Neubauer A., Fiebeler A., Graham D. K., O'Bryan J. P., Schmidt C. A., Barckow P., Serke S., Siegert W., Snodgrass H. R., Huhn D. Expression of axl, a transforming receptor tyrosine kinase, in normal and malignant hematopoiesis. Blood. 1994 Sep 15;84(6):1931–1941. [PubMed] [Google Scholar]
  27. Nichols E. J., Manger R., Hakomori S., Herscovics A., Rohrschneider L. R. Transformation by the v-fms oncogene product: role of glycosylational processing and cell surface expression. Mol Cell Biol. 1985 Dec;5(12):3467–3475. doi: 10.1128/mcb.5.12.3467. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. O'Bryan J. P., Frye R. A., Cogswell P. C., Neubauer A., Kitch B., Prokop C., Espinosa R., 3rd, Le Beau M. M., Earp H. S., Liu E. T. axl, a transforming gene isolated from primary human myeloid leukemia cells, encodes a novel receptor tyrosine kinase. Mol Cell Biol. 1991 Oct;11(10):5016–5031. doi: 10.1128/mcb.11.10.5016. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Ohashi K., Nagata K., Toshima J., Nakano T., Arita H., Tsuda H., Suzuki K., Mizuno K. Stimulation of sky receptor tyrosine kinase by the product of growth arrest-specific gene 6. J Biol Chem. 1995 Sep 29;270(39):22681–22684. doi: 10.1074/jbc.270.39.22681. [DOI] [PubMed] [Google Scholar]
  30. Poon B., Dixon D., Ellis L., Roth R. A., Rutter W. J., Wang L. H. Molecular basis of the activation of the tumorigenic potential of Gag-insulin receptor chimeras. Proc Natl Acad Sci U S A. 1991 Feb 1;88(3):877–881. doi: 10.1073/pnas.88.3.877. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Privalsky M. L., Bishop J. M. Subcellular localization of the v-erb-B protein, the product of a transforming gene of avian erythroblastosis virus. Virology. 1984 Jun;135(2):356–368. doi: 10.1016/0042-6822(84)90192-2. [DOI] [PubMed] [Google Scholar]
  32. Prywes R., Hoag J., Rosenberg N., Baltimore D. Protein stabilization explains the gag requirement for transformation of lymphoid cells by Abelson murine leukemia virus. J Virol. 1985 Apr;54(1):123–132. doi: 10.1128/jvi.54.1.123-132.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Reddy E. P., Smith M. J., Srinivasan A. Nucleotide sequence of Abelson murine leukemia virus genome: structural similarity of its transforming gene product to other onc gene products with tyrosine-specific kinase activity. Proc Natl Acad Sci U S A. 1983 Jun;80(12):3623–3627. doi: 10.1073/pnas.80.12.3623. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Rettenmier C. W., Roussel M. F., Quinn C. O., Kitchingman G. R., Look A. T., Sherr C. J. Transmembrane orientation of glycoproteins encoded by the v-fms oncogene. Cell. 1985 Apr;40(4):971–981. doi: 10.1016/0092-8674(85)90357-5. [DOI] [PubMed] [Google Scholar]
  35. Rodrigues G. A., Park M. Dimerization mediated through a leucine zipper activates the oncogenic potential of the met receptor tyrosine kinase. Mol Cell Biol. 1993 Nov;13(11):6711–6722. doi: 10.1128/mcb.13.11.6711. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Schlessinger J., Ullrich A. Growth factor signaling by receptor tyrosine kinases. Neuron. 1992 Sep;9(3):383–391. doi: 10.1016/0896-6273(92)90177-f. [DOI] [PubMed] [Google Scholar]
  37. Schmidt J. A., Beug H., Hayman M. J. Effects of inhibitors of glycoprotein processing on the synthesis and biological activity of the erb B oncogene. EMBO J. 1985 Jan;4(1):105–112. doi: 10.1002/j.1460-2075.1985.tb02323.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  38. Shinkai Y., Rathbun G., Lam K. P., Oltz E. M., Stewart V., Mendelsohn M., Charron J., Datta M., Young F., Stall A. M. RAG-2-deficient mice lack mature lymphocytes owing to inability to initiate V(D)J rearrangement. Cell. 1992 Mar 6;68(5):855–867. doi: 10.1016/0092-8674(92)90029-c. [DOI] [PubMed] [Google Scholar]
  39. Shinnick T. M., Lerner R. A., Sutcliffe J. G. Nucleotide sequence of Moloney murine leukaemia virus. Nature. 1981 Oct 15;293(5833):543–548. doi: 10.1038/293543a0. [DOI] [PubMed] [Google Scholar]
  40. Shu H. K., Chang C. M., Ravi L., Ling L., Castellano C. M., Walter E., Pelley R. J., Kung H. J. Modulation of erbB kinase activity and oncogenic potential by single point mutations in the glycine loop of the catalytic domain. Mol Cell Biol. 1994 Oct;14(10):6868–6878. doi: 10.1128/mcb.14.10.6868. [DOI] [PMC free article] [PubMed] [Google Scholar]
  41. Soman N. R., Correa P., Ruiz B. A., Wogan G. N. The TPR-MET oncogenic rearrangement is present and expressed in human gastric carcinoma and precursor lesions. Proc Natl Acad Sci U S A. 1991 Jun 1;88(11):4892–4896. doi: 10.1073/pnas.88.11.4892. [DOI] [PMC free article] [PubMed] [Google Scholar]
  42. 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]
  43. Stitt T. N., Conn G., Gore M., Lai C., Bruno J., Radziejewski C., Mattsson K., Fisher J., Gies D. R., Jones P. F. The anticoagulation factor protein S and its relative, Gas6, are ligands for the Tyro 3/Axl family of receptor tyrosine kinases. Cell. 1995 Feb 24;80(4):661–670. doi: 10.1016/0092-8674(95)90520-0. [DOI] [PubMed] [Google Scholar]
  44. Taylor I. C., Roy S., Yaswen P., Stampfer M. R., Varmus H. E. Mouse mammary tumors express elevated levels of RNA encoding the murine homology of SKY, a putative receptor tyrosine kinase. J Biol Chem. 1995 Mar 24;270(12):6872–6880. doi: 10.1074/jbc.270.12.6872. [DOI] [PubMed] [Google Scholar]
  45. Towbin H., Staehelin T., Gordon J. Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. Proc Natl Acad Sci U S A. 1979 Sep;76(9):4350–4354. doi: 10.1073/pnas.76.9.4350. [DOI] [PMC free article] [PubMed] [Google Scholar]
  46. Van Beveren C., Enami S., Curran T., Verma I. M. FBR murine osteosarcoma virus. II. Nucleotide sequence of the provirus reveals that the genome contains sequences acquired from two cellular genes. Virology. 1984 May;135(1):229–243. doi: 10.1016/0042-6822(84)90133-8. [DOI] [PubMed] [Google Scholar]
  47. Varnum B. C., Young C., Elliott G., Garcia A., Bartley T. D., Fridell Y. W., Hunt R. W., Trail G., Clogston C., Toso R. J. Axl receptor tyrosine kinase stimulated by the vitamin K-dependent protein encoded by growth-arrest-specific gene 6. Nature. 1995 Feb 16;373(6515):623–626. doi: 10.1038/373623a0. [DOI] [PubMed] [Google Scholar]
  48. Vu T. K., Hung D. T., Wheaton V. I., Coughlin S. R. Molecular cloning of a functional thrombin receptor reveals a novel proteolytic mechanism of receptor activation. Cell. 1991 Mar 22;64(6):1057–1068. doi: 10.1016/0092-8674(91)90261-v. [DOI] [PubMed] [Google Scholar]
  49. Wang L. H., Lin B., Jong S. M., Dixon D., Ellis L., Roth R. A., Rutter W. J. Activation of transforming potential of the human insulin receptor gene. Proc Natl Acad Sci U S A. 1987 Aug;84(16):5725–5729. doi: 10.1073/pnas.84.16.5725. [DOI] [PMC free article] [PubMed] [Google Scholar]
  50. Watanabe S. M., Rosenberg N. E., Witte O. N. A membrane-associated, carbohydrate-modified form of the v-abl protein that cannot be phosphorylated in vivo or in vitro. J Virol. 1984 Sep;51(3):620–627. doi: 10.1128/jvi.51.3.620-627.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  51. Wells A., Bishop J. M. Genetic determinants of neoplastic transformation by the retroviral oncogene v-erbB. Proc Natl Acad Sci U S A. 1988 Oct;85(20):7597–7601. doi: 10.1073/pnas.85.20.7597. [DOI] [PMC free article] [PubMed] [Google Scholar]

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