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. 1991 Dec;65(12):6478–6485. doi: 10.1128/jvi.65.12.6478-6485.1991

Carboxyl-terminal determinants of Abelson protein important for lymphoma induction.

K Parmar 1, R C Huebner 1, N Rosenberg 1
PMCID: PMC250691  PMID: 1942244

Abstract

The carboxyl-terminal region of the Abelson protein is not absolutely required for Abelson virus transformation. However, Abelson virus strains encoding proteins missing portions of this region have a reduced ability to transform lymphoid cells in vitro and in vivo. One such strain, called P90A, is unique in that P90A-injected mice almost always develop tumors containing highly oncogenic variants that encode new forms of Abelson protein. In this work, we have examined the mechanism by which these variants are generated and used the variants to identify carboxyl-terminal protein sequences important for the induction of Abelson disease. Analysis of mice injected with helper-free P90A virus stocks demonstrates that the variants are generated during viral replication in vivo, probably as a consequence of error-prone reverse transcription. The sequence of the P90A viral genome reveals that a 19-base deletion is responsible for synthesis of the truncated Abelson protein. As a consequence of this mutation, 167 carboxyl-terminal amino acids normally found in the wild-type protein have been replaced by 33 amino acids derived from an alternative reading frame. Site-directed mutants show that the combination of the deletion and the P90A carboxyl terminus is required for the generation of variants. Thus, the particular structure of the P90A protein, not the specific residues lost or gained, alters the transforming potential of the Abelson protein. Finally, the sequence of the variants encoding smaller Abelson proteins reveals that as few as 452 v-abl-encoded amino acids are required for rapid induction of Abelson disease.

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Selected References

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

  1. Bassin R. H., Tuttle N., Fischinger P. J. Rapid cell culture assay technic for murine leukaemia viruses. Nature. 1971 Feb 19;229(5286):564–566. doi: 10.1038/229564b0. [DOI] [PubMed] [Google Scholar]
  2. Bergold P. J., Blumenthal J. A., D'Andrea E., Snyder H. W., Lederman L., Silverstone A., Nguyen H., Besmer P. Nucleic acid sequence and oncogenic properties of the HZ2 feline sarcoma virus v-abl insert. J Virol. 1987 Apr;61(4):1193–1202. doi: 10.1128/jvi.61.4.1193-1202.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Canaani E., Robbins K. C., Aaronson S. A. The transforming gene of Moloney murine sarcoma virus. Nature. 1979 Nov 22;282(5737):378–383. doi: 10.1038/282378a0. [DOI] [PubMed] [Google Scholar]
  4. Engelman A., Rosenberg N. Isolation of temperature-sensitive Abelson virus mutants by site-directed mutagenesis. Proc Natl Acad Sci U S A. 1987 Nov;84(22):8021–8025. doi: 10.1073/pnas.84.22.8021. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Engelman A., Rosenberg N. Temperature-sensitive mutants of Abelson murine leukemia virus deficient in protein tyrosine kinase activity. J Virol. 1990 Sep;64(9):4242–4251. doi: 10.1128/jvi.64.9.4242-4251.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Goff S. P., Gilboa E., Witte O. N., Baltimore D. Structure of the Abelson murine leukemia virus genome and the homologous cellular gene: studies with cloned viral DNA. Cell. 1980 Dec;22(3):777–785. doi: 10.1016/0092-8674(80)90554-1. [DOI] [PubMed] [Google Scholar]
  7. Goff S. P., Tabin C. J., Wang J. Y., Weinberg R., Baltimore D. Transfection of fibroblasts by cloned Abelson murine leukemia virus DNA and recovery of transmissible virus by recombination with helper virus. J Virol. 1982 Jan;41(1):271–285. doi: 10.1128/jvi.41.1.271-285.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Green P. L., Kaehler D. A., Risser R. Cell transformation and tumor induction by Abelson murine leukemia virus in the absence of helper virus. Proc Natl Acad Sci U S A. 1987 Aug;84(16):5932–5936. doi: 10.1073/pnas.84.16.5932. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Hanafusa H., Halpern C. C., Buchhagen D. L., Kawai S. Recovery of avian sarcoma virus from tumors induced by transformation-defective mutants. J Exp Med. 1977 Dec 1;146(6):1735–1747. doi: 10.1084/jem.146.6.1735. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Hanks S. K., Quinn A. M., Hunter T. The protein kinase family: conserved features and deduced phylogeny of the catalytic domains. Science. 1988 Jul 1;241(4861):42–52. doi: 10.1126/science.3291115. [DOI] [PubMed] [Google Scholar]
  11. Henkemeyer M. J., Bennett R. L., Gertler F. B., Hoffmann F. M. DNA sequence, structure, and tyrosine kinase activity of the Drosophila melanogaster Abelson proto-oncogene homolog. Mol Cell Biol. 1988 Feb;8(2):843–853. doi: 10.1128/mcb.8.2.843. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. 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]
  13. Kipreos E. T., Lee G. J., Wang J. Y. Isolation of temperature-sensitive tyrosine kinase mutants of v-abl oncogene by screening with antibodies for phosphotyrosine. Proc Natl Acad Sci U S A. 1987 Mar;84(5):1345–1349. doi: 10.1073/pnas.84.5.1345. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. 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]
  15. Lobel L. I., Murphy J. E., Goff S. P. The palindromic LTR-LTR junction of Moloney murine leukemia virus is not an efficient substrate for proviral integration. J Virol. 1989 Jun;63(6):2629–2637. doi: 10.1128/jvi.63.6.2629-2637.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Loenen W. A., Brammar W. J. A bacteriophage lambda vector for cloning large DNA fragments made with several restriction enzymes. Gene. 1980 Aug;10(3):249–259. doi: 10.1016/0378-1119(80)90054-2. [DOI] [PubMed] [Google Scholar]
  17. Markowitz D., Goff S., Bank A. Construction and use of a safe and efficient amphotropic packaging cell line. Virology. 1988 Dec;167(2):400–406. [PubMed] [Google Scholar]
  18. Murtagh K., Skladany G., Hoag J., Rosenberg N. Abelson murine leukemia virus variants with increased oncogenic potential. J Virol. 1986 Nov;60(2):599–606. doi: 10.1128/jvi.60.2.599-606.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Pendergast A. M., Traugh J. A., Witte O. N. Normal cellular and transformation-associated abl proteins share common sites for protein kinase C phosphorylation. Mol Cell Biol. 1987 Dec;7(12):4280–4289. doi: 10.1128/mcb.7.12.4280. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Ponticelli A. S., Whitlock C. A., Rosenberg N., Witte O. N. In vivo tyrosine phosphorylations of the Abelson virus transforming protein are absent in its normal cellular homolog. Cell. 1982 Jul;29(3):953–960. doi: 10.1016/0092-8674(82)90458-5. [DOI] [PubMed] [Google Scholar]
  21. Prywes R., Foulkes J. G., Baltimore D. The minimum transforming region of v-abl is the segment encoding protein-tyrosine kinase. J Virol. 1985 Apr;54(1):114–122. doi: 10.1128/jvi.54.1.114-122.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Prywes R., Foulkes J. G., Rosenberg N., Baltimore D. Sequences of the A-MuLV protein needed for fibroblast and lymphoid cell transformation. Cell. 1983 Sep;34(2):569–579. doi: 10.1016/0092-8674(83)90389-6. [DOI] [PubMed] [Google Scholar]
  23. 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]
  24. Ramsay G. M., Hayman M. J. Isolation and biochemical characterization of partially transformation-defective mutants of avian myelocytomatosis virus strain MC29: localization of the mutation to the myc domain of the 110,000-dalton gag-myc polyprotein. J Virol. 1982 Mar;41(3):745–753. doi: 10.1128/jvi.41.3.745-753.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. 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]
  26. Reynolds F. H., Jr, Sacks T. L., Deobagkar D. N., Stephenson J. R. Cells nonproductively transformed by Abelson murine leukemia virus express a high molecular weight polyprotein containing structural and nonstructural components. Proc Natl Acad Sci U S A. 1978 Aug;75(8):3974–3978. doi: 10.1073/pnas.75.8.3974. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Reynolds F. H., Jr, Van de Ven W. J., Stephenson J. R. Abelson murine leukemia virus transformation-defective mutants with impaired P120-associated protein kinase activity. J Virol. 1980 Nov;36(2):374–386. doi: 10.1128/jvi.36.2.374-386.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Rimm D. L., Horness D., Kucera J., Blattner F. R. Construction of coliphage lambda Charon vectors with BamHI cloning sites. Gene. 1980 Dec;12(3-4):301–309. doi: 10.1016/0378-1119(80)90113-4. [DOI] [PubMed] [Google Scholar]
  29. Rosenberg N. E., Clark D. R., Witte O. N. Abelson murine leukemia virus mutants deficient in kinase activity and lymphoid cell transformation. J Virol. 1980 Dec;36(3):766–774. doi: 10.1128/jvi.36.3.766-774.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Rosenberg N., Witte O. N. Abelson murine leukemia virus mutants with alterations in the virus-specific P120 molecule. J Virol. 1980 Jan;33(1):340–348. doi: 10.1128/jvi.33.1.340-348.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Rosenberg N., Witte O. N. The viral and cellular forms of the Abelson (abl) oncogene. Adv Virus Res. 1988;35:39–81. doi: 10.1016/s0065-3527(08)60708-3. [DOI] [PubMed] [Google Scholar]
  32. Saiki R. K., Gelfand D. H., Stoffel S., Scharf S. J., Higuchi R., Horn G. T., Mullis K. B., Erlich H. A. Primer-directed enzymatic amplification of DNA with a thermostable DNA polymerase. Science. 1988 Jan 29;239(4839):487–491. doi: 10.1126/science.2448875. [DOI] [PubMed] [Google Scholar]
  33. 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]
  34. Scher C. D., Siegler R. Direct transformation of 3T3 cells by Abelson murine leukaemia virus. Nature. 1975 Feb 27;253(5494):729–731. doi: 10.1038/253729a0. [DOI] [PubMed] [Google Scholar]
  35. Schwartzberg P. L., Goff S. P., Robertson E. J. Germ-line transmission of a c-abl mutation produced by targeted gene disruption in ES cells. Science. 1989 Nov 10;246(4931):799–803. doi: 10.1126/science.2554496. [DOI] [PubMed] [Google Scholar]
  36. Shoemaker C., Goff S., Gilboa E., Paskind M., Mitra S. W., Baltimore D. Structure of a cloned circular Moloney murine leukemia virus DNA molecule containing an inverted segment: implications for retrovirus integration. Proc Natl Acad Sci U S A. 1980 Jul;77(7):3932–3936. doi: 10.1073/pnas.77.7.3932. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. Shtivelman E., Lifshitz B., Gale R. P., Roe B. A., Canaani E. Alternative splicing of RNAs transcribed from the human abl gene and from the bcr-abl fused gene. Cell. 1986 Oct 24;47(2):277–284. doi: 10.1016/0092-8674(86)90450-2. [DOI] [PubMed] [Google Scholar]
  38. Takemori T., Miyazoe I., Shirasawa T., Taniguchi M., Graf T. A temperature-sensitive mutant of Abelson murine leukemia virus confers inducibility of IgM expression to transformed lymphoid cells. EMBO J. 1987 Apr;6(4):951–956. doi: 10.1002/j.1460-2075.1987.tb04844.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  39. Van Etten R. A., Jackson P., Baltimore D. The mouse type IV c-abl gene product is a nuclear protein, and activation of transforming ability is associated with cytoplasmic localization. Cell. 1989 Aug 25;58(4):669–678. doi: 10.1016/0092-8674(89)90102-5. [DOI] [PubMed] [Google Scholar]
  40. Wang J. Y., Baltimore D. Localization of tyrosine kinase-coding region in v-abl oncogene by the expression of v-abl-encoded proteins in bacteria. J Biol Chem. 1985 Jan 10;260(1):64–71. [PubMed] [Google Scholar]
  41. 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]
  42. Watanabe S. M., Witte O. N. Site-directed deletions of Abelson murine leukemia virus define 3' sequences essential for transformation and lethality. J Virol. 1983 Mar;45(3):1028–1036. doi: 10.1128/jvi.45.3.1028-1036.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  43. Wigler M., Sweet R., Sim G. K., Wold B., Pellicer A., Lacy E., Maniatis T., Silverstein S., Axel R. Transformation of mammalian cells with genes from procaryotes and eucaryotes. Cell. 1979 Apr;16(4):777–785. doi: 10.1016/0092-8674(79)90093-x. [DOI] [PubMed] [Google Scholar]
  44. Witte O. N., Dasgupta A., Baltimore D. Abelson murine leukaemia virus protein is phosphorylated in vitro to form phosphotyrosine. Nature. 1980 Feb 28;283(5750):826–831. doi: 10.1038/283826a0. [DOI] [PubMed] [Google Scholar]
  45. Witte O. N., Goff S., Rosenberg N., Baltimore D. A transformation-defective mutant of Abelson murine leukemia virus lacks protein kinase activity. Proc Natl Acad Sci U S A. 1980 Aug;77(8):4993–4997. doi: 10.1073/pnas.77.8.4993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  46. Witte O. N., Rosenberg N., Paskind M., Shields A., Baltimore D. Identification of an Abelson murine leukemia virus-encoded protein present in transformed fibroblast and lymphoid cells. Proc Natl Acad Sci U S A. 1978 May;75(5):2488–2492. doi: 10.1073/pnas.75.5.2488. [DOI] [PMC free article] [PubMed] [Google Scholar]
  47. Ziegler S. F., Whitlock C. A., Goff S. P., Gifford A., Witte O. N. Lethal effect of the Abelson murine leukemia virus transforming gene product. Cell. 1981 Dec;27(3 Pt 2):477–486. doi: 10.1016/0092-8674(81)90389-5. [DOI] [PubMed] [Google Scholar]
  48. Zoller M. J., Smith M. Oligonucleotide-directed mutagenesis: a simple method using two oligonucleotide primers and a single-stranded DNA template. DNA. 1984 Dec;3(6):479–488. doi: 10.1089/dna.1.1984.3.479. [DOI] [PubMed] [Google Scholar]

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