Differential complementation of Bcr-Abl point mutants with c-Myc

Science

Volumn 264, Issue 5157, 1994, Pages 424-426

  Afar, Daniel E H ^a   Goga, Andrei ^a   McLaughlin, Jami ^a   Witte, Owen N ^a   Sawyers, Charles L ^a  

^a UNIVERSITY OF CALIFORNIA   (United States)

Author keywords

[No Author keywords available]

Indexed keywords

PROTEIN TYROSINE KINASE;

ARTICLE; AUTOPHOSPHORYLATION; BINDING SITE; CELL TRANSFORMATION; ONCOGENE; ONCOGENE C MYC; POINT MUTATION; PRIORITY JOURNAL; SIGNAL TRANSDUCTION;

EID: 0028286140     PISSN: 00368075     EISSN: None     Source Type: Journal    
DOI: 10.1126/science.8153630     Document Type: Article

References (39)

1. B. Vogelstein and K. W. Kinzler, Trends Genet. 9, 138 (1993).
2. Shtivelman, B. Lifshitz, R. P. Gale, E. Canaani, Nature 315, 550 (1985); S. C. Clark et al., Science 239, 775 (1988)
3. Shtivelman, B. Lifshitz, R. P. Gale, E. Canaani, Nature 315, 550 (1985); S. C. Clark et al., Science 239, 775 (1988)
4. A. M. Pendergast, M. L. Gishizky, M. H. Havlik, O. N. Witte, Mol. Cell. Biol. 13, 1728 (1993).
5. A. M. Pendergast et al., Cell 75, 175 (1993).
6. E. J. Lowenstein et al., ibid. 70, 431 (1992); K. Baltensperger et al., Science 260, 1950 91993); L. Buday and J. Downward, Cell 73, 611 (1993); S. E. Egan et al., Nature 363, 45 (1993); N. W. Gale, S. Kaplan, E. J. Lowenstein, J. Schlessinger, D. Bar-Sagi, ibid., p. 88; N. Li et al., ibid., p. 85; J. P. Olivier et al., Cell 73, 179 (1993); M. Rozakis-Adcock, R. Fernley, J. Wade, T. Pawson, D. Bowtell, Nature 363, 83 (1993); A. Simon, G. S. Dodson, G. M. Rubin, Cell 73, 169 (1993); E. Y. Skolnik et al., Science 260, 1953 (1993).
7. E. J. Lowenstein et al., ibid. 70, 431 (1992); K. Baltensperger et al., Science 260, 1950 91993); L. Buday and J. Downward, Cell 73, 611 (1993); S. E. Egan et al., Nature 363, 45 (1993); N. W. Gale, S. Kaplan, E. J. Lowenstein, J. Schlessinger, D. Bar-Sagi, ibid., p. 88; N. Li et al., ibid., p. 85; J. P. Olivier et al., Cell 73, 179 (1993); M. Rozakis-Adcock, R. Fernley, J. Wade, T. Pawson, D. Bowtell, Nature 363, 83 (1993); A. Simon, G. S. Dodson, G. M. Rubin, Cell 73, 169 (1993); E. Y. Skolnik et al., Science 260, 1953 (1993).
8. E. J. Lowenstein et al., ibid. 70, 431 (1992); K. Baltensperger et al., Science 260, 1950 91993); L. Buday and J. Downward, Cell 73, 611 (1993); S. E. Egan et al., Nature 363, 45 (1993); N. W. Gale, S. Kaplan, E. J. Lowenstein, J. Schlessinger, D. Bar-Sagi, ibid., p. 88; N. Li et al., ibid., p. 85; J. P. Olivier et al., Cell 73, 179 (1993); M. Rozakis-Adcock, R. Fernley, J. Wade, T. Pawson, D. Bowtell, Nature 363, 83 (1993); A. Simon, G. S. Dodson, G. M. Rubin, Cell 73, 169 (1993); E. Y. Skolnik et al., Science 260, 1953 (1993).
9. E. J. Lowenstein et al., ibid. 70, 431 (1992); K. Baltensperger et al., Science 260, 1950 91993); L. Buday and J. Downward, Cell 73, 611 (1993); S. E. Egan et al., Nature 363, 45 (1993); N. W. Gale, S. Kaplan, E. J. Lowenstein, J. Schlessinger, D. Bar-Sagi, ibid., p. 88; N. Li et al., ibid., p. 85; J. P. Olivier et al., Cell 73, 179 (1993); M. Rozakis-Adcock, R. Fernley, J. Wade, T. Pawson, D. Bowtell, Nature 363, 83 (1993); A. Simon, G. S. Dodson, G. M. Rubin, Cell 73, 169 (1993); E. Y. Skolnik et al., Science 260, 1953 (1993).
10. E. J. Lowenstein et al., ibid. 70, 431 (1992); K. Baltensperger et al., Science 260, 1950 91993); L. Buday and J. Downward, Cell 73, 611 (1993); S. E. Egan et al., Nature 363, 45 (1993); N. W. Gale, S. Kaplan, E. J. Lowenstein, J. Schlessinger, D. Bar-Sagi, ibid., p. 88; N. Li et al., ibid., p. 85; J. P. Olivier et al., Cell 73, 179 (1993); M. Rozakis-Adcock, R. Fernley, J. Wade, T. Pawson, D. Bowtell, Nature 363, 83 (1993); A. Simon, G. S. Dodson, G. M. Rubin, Cell 73, 169 (1993); E. Y. Skolnik et al., Science 260, 1953 (1993).
11. E. J. Lowenstein et al., ibid. 70, 431 (1992); K. Baltensperger et al., Science 260, 1950 91993); L. Buday and J. Downward, Cell 73, 611 (1993); S. E. Egan et al., Nature 363, 45 (1993); N. W. Gale, S. Kaplan, E. J. Lowenstein, J. Schlessinger, D. Bar-Sagi, ibid., p. 88; N. Li et al., ibid., p. 85; J. P. Olivier et al., Cell 73, 179 (1993); M. Rozakis-Adcock, R. Fernley, J. Wade, T. Pawson, D. Bowtell, Nature 363, 83 (1993); A. Simon, G. S. Dodson, G. M. Rubin, Cell 73, 169 (1993); E. Y. Skolnik et al., Science 260, 1953 (1993).
12. E. J. Lowenstein et al., ibid. 70, 431 (1992); K. Baltensperger et al., Science 260, 1950 91993); L. Buday and J. Downward, Cell 73, 611 (1993); S. E. Egan et al., Nature 363, 45 (1993); N. W. Gale, S. Kaplan, E. J. Lowenstein, J. Schlessinger, D. Bar-Sagi, ibid., p. 88; N. Li et al., ibid., p. 85; J. P. Olivier et al., Cell 73, 179 (1993); M. Rozakis-Adcock, R. Fernley, J. Wade, T. Pawson, D. Bowtell, Nature 363, 83 (1993); A. Simon, G. S. Dodson, G. M. Rubin, Cell 73, 169 (1993); E. Y. Skolnik et al., Science 260, 1953 (1993).
13. E. J. Lowenstein et al., ibid. 70, 431 (1992); K. Baltensperger et al., Science 260, 1950 91993); L. Buday and J. Downward, Cell 73, 611 (1993); S. E. Egan et al., Nature 363, 45 (1993); N. W. Gale, S. Kaplan, E. J. Lowenstein, J. Schlessinger, D. Bar-Sagi, ibid., p. 88; N. Li et al., ibid., p. 85; J. P. Olivier et al., Cell 73, 179 (1993); M. Rozakis-Adcock, R. Fernley, J. Wade, T. Pawson, D. Bowtell, Nature 363, 83 (1993); A. Simon, G. S. Dodson, G. M. Rubin, Cell 73, 169 (1993); E. Y. Skolnik et al., Science 260, 1953 (1993).
14. E. J. Lowenstein et al., ibid. 70, 431 (1992); K. Baltensperger et al., Science 260, 1950 91993); L. Buday and J. Downward, Cell 73, 611 (1993); S. E. Egan et al., Nature 363, 45 (1993); N. W. Gale, S. Kaplan, E. J. Lowenstein, J. Schlessinger, D. Bar-Sagi, ibid., p. 88; N. Li et al., ibid., p. 85; J. P. Olivier et al., Cell 73, 179 (1993); M. Rozakis-Adcock, R. Fernley, J. Wade, T. Pawson, D. Bowtell, Nature 363, 83 (1993); A. Simon, G. S. Dodson, G. M. Rubin, Cell 73, 169 (1993); E. Y. Skolnik et al., Science 260, 1953 (1993).
15. E. J. Lowenstein et al., ibid. 70, 431 (1992); K. Baltensperger et al., Science 260, 1950 91993); L. Buday and J. Downward, Cell 73, 611 (1993); S. E. Egan et al., Nature 363, 45 (1993); N. W. Gale, S. Kaplan, E. J. Lowenstein, J. Schlessinger, D. Bar-Sagi, ibid., p. 88; N. Li et al., ibid., p. 85; J. P. Olivier et al., Cell 73, 179 (1993); M. Rozakis-Adcock, R. Fernley, J. Wade, T. Pawson, D. Bowtell, Nature 363, 83 (1993); A. Simon, G. S. Dodson, G. M. Rubin, Cell 73, 169 (1993); E. Y. Skolnik et al., Science 260, 1953 (1993).
16. C. A. Koch, D. Anderson, M. F. Moran, C. Ellis, T. Pawson, Science 252, 668 (1991) Abbreviations for the amino acid residues are as follows; A, Ala; C, Cys; D, Asp; E, Glu; F, Phe; G, Gly; H, His; I, Ile; K, Lys; L, Leu; M, Met; N, Asn; P, Pro; Q, Gln; R, Arg; S, Ser; T, Thr; V, Val; W, Trp; and Y, Tyr.
17. G. Waksman et al., Nature 358, 646 (1992).
18. M. F. Verderame, J. M. Kaplan, H. E. Varmus, J. Virol. 63, 338 (1989); H. Hirai and H. E. Varmus, Genes Dev. 4, 2342 (1990); B. J. Mayer, P. K. Jackson, R. A. Van Etten, D. Baltimore, Mol. Cell. Biol. 12, 609 (1992).
19. M. F. Verderame, J. M. Kaplan, H. E. Varmus, J. Virol. 63, 338 (1989); H. Hirai and H. E. Varmus, Genes Dev. 4, 2342 (1990); B. J. Mayer, P. K. Jackson, R. A. Van Etten, D. Baltimore, Mol. Cell. Biol. 12, 609 (1992).
20. M. F. Verderame, J. M. Kaplan, H. E. Varmus, J. Virol. 63, 338 (1989); H. Hirai and H. E. Varmus, Genes Dev. 4, 2342 (1990); B. J. Mayer, P. K. Jackson, R. A. Van Etten, D. Baltimore, Mol. Cell. Biol. 12, 609 (1992).
21. 552 in the SH2 domain of P185 with Leu was 5′-TATCCGCTGAGCAGCGGGATCAATGGCAGCTTCTTGGTGCTTGAGAGTGAGAGCAGTCC-3′, where the underlined sequence replaced the WT codon of CGT. The downstream primer used was 3′-CGTAAACCTCATAACGAAACCTTCGAACGC-5′. Sequences of c-abl were used as a template to PCR-amplify the mutant SH2 domain. The PCR product was subcloned into c-abl pBluescript (Stratagene) and analyzed by dideoxy DNA sequencing (Sequenase, United States Biochemical) to verify the presence of the point mutation. The FLVRES mutation was then reconstructed into Bcr-Abl by ligation of a Kpn I and Hind III digest of c-abl FLVRES mutant to an Eco RI and Kpn I digest of P185 bcr-abl. The P185 FLVRES mutant was then subcloned into the pSRαMSVtkneo retroviral vector (10) as an Eco RI and Hind III fragment. The autophosphorylation mutant P185 Y813F was generated by the recombination of the 5′ end of P185 bcr-abl with a Kpn I and Hind III fragment of P210 bcr-abl 1294F (3). To generate the double mutant P185 L552 and F813, P185 Y813F was digested with Bsr GI and Hind III and recombined with the 5′ end of P185 R552L.
22. A. J. Muller et al., Mol. Cell. Biol. 11, 1785 (1991).
23. Possible identities of p62 and p65 include previously identified molecules such as the Ras-guanosine triphosphatase activating protein-associated p62, the protein tyrosine phosphatase Syp, and the p66 isoform of Shc. G. Wong et al., Cell 69, 551 (1992); G.-S. Feng, C.-C. Hui, T. Pawson, Science 259, 1607 (1993); J. McGlade, A. Cheng, G. Pelicci, P. G. Pelicci, T. Pawson, Proc. Natl. Acad. Sci. U.S.A. 89, 8869 (1992).
24. Possible identities of p62 and p65 include previously identified molecules such as the Ras-guanosine triphosphatase activating protein-associated p62, the protein tyrosine phosphatase Syp, and the p66 isoform of Shc. G. Wong et al., Cell 69, 551 (1992); G.-S. Feng, C.-C. Hui, T. Pawson, Science 259, 1607 (1993); J. McGlade, A. Cheng, G. Pelicci, P. G. Pelicci, T. Pawson, Proc. Natl. Acad. Sci. U.S.A. 89, 8869 (1992).
25. Possible identities of p62 and p65 include previously identified molecules such as the Ras-guanosine triphosphatase activating protein-associated p62, the protein tyrosine phosphatase Syp, and the p66 isoform of Shc. G. Wong et al., Cell 69, 551 (1992); G.-S. Feng, C.-C. Hui, T. Pawson, Science 259, 1607 (1993); J. McGlade, A. Cheng, G. Pelicci, P. G. Pelicci, T. Pawson, Proc. Natl. Acad. Sci. U.S.A. 89, 8869 (1992).
26. A fraction of Grb-2 molecules was found to undergo a mobility shift within cells expressing all P185 forms, excluding cells expressing P185 Y177F. Immunoblot analysis with antibody to phosphotyrosine revealed that the upper band was tyrosine-phosphorylated Grb-2, which may be the result of Bcr-Abl overexpression in 293T cells.
27. T. Lugo and O. N. Witte, Mol. Cell. Biol. 9, 1263 (1989).
28. C. L. Sawyers, W. Callahan, O. N. Witte, Cell 70, 901 (1992).
29. Three independent transformed colonies were isolated and expanded in tissue culture. Bcr-Abl expression, intracellular phosphotyrosine concentrations, and DNA sequencing confirmed the presence of the FLVRES mutation. Protein immunoblot analysis of transformed colonies with antibody to phosphotyrosine gave results indistinguishable from those shown in Fig. 1.
30. M. F. Roussel, J. L. Cleveland, S. A. Shurtleff, C. J. Sherr, Nature 353, 361 (1991).
31. J. E. DeClue, K. Zhang, P. Redford, W. C. Vass, D. R. Lowy, Mol. Cell. Biol. 11, 2819 (1991); S. M. Thomas, M. DeMarco, G. D'Arcangelo, S. Halegoua, J. S. Brugge, Cell 68, 1031 (1992); K. W. Wood, C. Sarnecki, T. M. Roberts, J. Blenis, ibid., p. 1041.
32. J. E. DeClue, K. Zhang, P. Redford, W. C. Vass, D. R. Lowy, Mol. Cell. Biol. 11, 2819 (1991); S. M. Thomas, M. DeMarco, G. D'Arcangelo, S. Halegoua, J. S. Brugge, Cell 68, 1031 (1992); K. W. Wood, C. Sarnecki, T. M. Roberts, J. Blenis, ibid., p. 1041.
33. J. E. DeClue, K. Zhang, P. Redford, W. C. Vass, D. R. Lowy, Mol. Cell. Biol. 11, 2819 (1991); S. M. Thomas, M. DeMarco, G. D'Arcangelo, S. Halegoua, J. S. Brugge, Cell 68, 1031 (1992); K. W. Wood, C. Sarnecki, T. M. Roberts, J. Blenis, ibid., p. 1041.
34. C. L. Sawyers, J. McLaughlin, M. Havlik, O. N. Witte, in preparation.
35. L. Puil et al., EMBO J., in press.
36. R. B. DuBridge et al., Mol. Cell. Biol. 7, 379 (1987).
37. J. B. Konopka and O. N. Witte, ibid. 5, 3116 (1985).
38. 4 cells per 6-cm dish. The samples were plated in duplicate in medium containing fetal calf serum (20%). Colonies equal to and larger than 0.5 mm in size were counted for 2 to 3 weeks after growth in soft agar. The numbers of colonies grown in soft agar represent averages from five independent experiments for cells expressing WT P185. Colonies expressing mutant P185 proteins were averaged from two to three independent experiments. Large colonies (L) represent sizes ranging from 0.5 to 3.0 mm in diameter. Small colonies (S) represent visually detectable colonies ≤0.5 mm in diameter. G418-resistant populations expressing P185 proteins were superinfected with retrovirus containing the neo gene or retrovirus containing c-myc.
39. We thank A. M. Pendergast for providing the Grb-2 binding mutant of Bcr-Abl (P185 Y177F); S. Smale, D. Black, A. Berk, L. Zipursky, D. Saffran, and L. Cohen for critical review of the manuscript; S. Quan and J. C. White for photography; and J. Shimaoka for assistance in preparation of the manuscript. D.E.H.A. is a fellow of the Medical Research Council of Canada. A.G. is a fellow of the UCLA medical scientist training program. O.N.W. is an Investigator of the Howard Hughes Medical Institute. Supported by the Physician Scientist Award CA 01551 from the National Cancer Institute (C.L.S.) and by grant CA 53867 from the National Institutes of Health (O.N.W.).