Maintenance of human T cell anergy: Blocking of IL-2 gene transcription by activated Rap1

Science

Volumn 278, Issue 5335, 1997, Pages 124-128

  Boussiotis, Vassiliki A ^a,b   Freeman, Gordon J ^a,b   Berezovskaya, Alla ^a,b   Barber, Dwayne L ^c   Nadler, Lee M ^a,b  

^a DANA FARBER CANCER INSTITUTE   (United States)

^b BRIGHAM AND WOMEN S HOSPITAL   (United States)

^c PRINCESS MARGARET HOSPITAL   (Canada)

Author keywords

[No Author keywords available]

Indexed keywords

GUANOSINE TRIPHOSPHATE; INTERLEUKIN 2; T LYMPHOCYTE RECEPTOR;

ANIMAL CELL; ANTIGEN RECOGNITION; ANTIGEN SPECIFICITY; ARTICLE; CLONAL ANERGY; HUMAN; HUMAN CELL; MOUSE; NONHUMAN; PRIORITY JOURNAL; PROTEIN EXPRESSION; T LYMPHOCYTE ACTIVATION; TRANSCRIPTION REGULATION;

ADAPTOR PROTEINS, SIGNAL TRANSDUCING; ANTIGENS, CD28; CLONAL ANERGY; GENE EXPRESSION REGULATION; GTP-BINDING PROTEINS; GUANINE NUCLEOTIDE EXCHANGE FACTORS; GUANOSINE TRIPHOSPHATE; HUMANS; INTERLEUKIN-2; JURKAT CELLS; NUCLEAR PROTEINS; PHOSPHORYLATION; PROTEIN-TYROSINE KINASES; PROTEINS; PROTO-ONCOGENE PROTEINS; PROTO-ONCOGENE PROTEINS C-CBL; PROTO-ONCOGENE PROTEINS C-FYN; RAP GTP-BINDING PROTEINS; RAS GUANINE NUCLEOTIDE EXCHANGE FACTORS; RAS PROTEINS; RECEPTORS, ANTIGEN, T-CELL; RECOMBINANT FUSION PROTEINS; SIGNAL TRANSDUCTION; SRC HOMOLOGY DOMAINS; T-LYMPHOCYTES; TRANSCRIPTION, GENETIC; TRANSFECTION; UBIQUITIN-PROTEIN LIGASES;

ANIMALIA;

EID: 0030812812     PISSN: 00368075     EISSN: None     Source Type: Journal    
DOI: 10.1126/science.278.5335.124     Document Type: Article

References (82)

1. M. K. Jenkins and R. H. Schwartz, J. Exp. Med. 165, 302 (1987); H. Quill and R. H. Schwartz, J. Immunol. 138, 3704 (1987); R. H. Schwartz, Science 248, 1349 (1990); J. Exp. Med. 184, 1 (1996).
2. M. K. Jenkins and R. H. Schwartz, J. Exp. Med. 165, 302 (1987); H. Quill and R. H. Schwartz, J. Immunol. 138, 3704 (1987); R. H. Schwartz, Science 248, 1349 (1990); J. Exp. Med. 184, 1 (1996).
3. M. K. Jenkins and R. H. Schwartz, J. Exp. Med. 165, 302 (1987); H. Quill and R. H. Schwartz, J. Immunol. 138, 3704 (1987); R. H. Schwartz, Science 248, 1349 (1990); J. Exp. Med. 184, 1 (1996).
4. M. K. Jenkins and R. H. Schwartz, J. Exp. Med. 165, 302 (1987); H. Quill and R. H. Schwartz, J. Immunol. 138, 3704 (1987); R. H. Schwartz, Science 248, 1349 (1990); J. Exp. Med. 184, 1 (1996).
5. K. Migita et al., J. Immunol. 155, 5083 (1995).
6. W. Li, C. D. Whaley, A. Mondino, D. L. Mueller, Science 271, 1272 (1996); P. E. Fields, T. F. Gajewski, F. W. Fitch, ibid., p. 1276.
7. W. Li, C. D. Whaley, A. Mondino, D. L. Mueller, Science 271, 1272 (1996); P. E. Fields, T. F. Gajewski, F. W. Fitch, ibid., p. 1276.
8. D. R. DeSilva, W. S. Feeser, E. J. Tancula, P. A. Scherle, J. Exp. Med. 183, 2017 (1996); S. M. Kang et al., Science 257, 1134 (1992).
9. D. R. DeSilva, W. S. Feeser, E. J. Tancula, P. A. Scherle, J. Exp. Med. 183, 2017 (1996); S. M. Kang et al., Science 257, 1134 (1992).
10. T. F. Gajewski, D. Qian, P. Fields, F. W. Fitch, Proc. Natl. Acad. Sci. U.S.A. 91, 38 (1994); T. F. Gajewski, P. Fields, F. W. Fitch, Eur. J. Immunol. 25, 1836 (1995).
11. T. F. Gajewski, D. Qian, P. Fields, F. W. Fitch, Proc. Natl. Acad. Sci. U.S.A. 91, 38 (1994); T. F. Gajewski, P. Fields, F. W. Fitch, Eur. J. Immunol. 25, 1836 (1995).
12. V. A. Boussiotis et al., Science. 266, 1039 (1994); V. A. Boussiotis et al., J. Exp. Med. 184, 365 (1996).
13. V. A. Boussiotis et al., Science. 266, 1039 (1994); V. A. Boussiotis et al., J. Exp. Med. 184, 365 (1996).
14. HLA-DR7 alloantigen-specific T cell clones (6) were cultured with the indicated NIH-3T3 stimulators in a primary culture for 24 hours, separated from NIH-3T3 transfectants by Percoll gradient centrifugation, and rested in media for 48 hours before biochemical analysis. For the kinetics of Cbl tyrosine phosphorylation in anergy, T cell clones were cultured with anergizing stimulators (t-DR7) for the indicated time intervals and then separated and rested in media as above.
15. G. Panchamoorthy et al., J. Biol. Chem. 271, 3187 (1996); T. Tezuka et al., J. Exp. Med. 183, 675 (1996).
16. G. Panchamoorthy et al., J. Biol. Chem. 271, 3187 (1996); T. Tezuka et al., J. Exp. Med. 183, 675 (1996).
17. T. Fukazawa et al., J. Biol. Chem. 270, 19141 (1995).
18. J. A. Donovan, R. L. Wange, W. Y. Langdon, L. E. Samelson, ibid. 269, 22921 (1994).
19. V. A. Boussiotis, G. J. Freeman, A. Berezovskaya, D. L. Barber, L. M. Nadler, unpublished results.
20. K. A. Reedquist et al., J. Biol. Chem. 271, 8453 (1996).
21. S. Sawasdikosol, K. S. Ravichandran, K. K. Lee, J. H. Chang, S. J. Burakoff, ibid. 270, 2893 (1995); L. Buday, A. Khwaja, S. Sipeki, A. Farago, J. Downward, ibid. 271, 6159 (1996).
22. S. Sawasdikosol, K. S. Ravichandran, K. K. Lee, J. H. Chang, S. J. Burakoff, ibid. 270, 2893 (1995); L. Buday, A. Khwaja, S. Sipeki, A. Farago, J. Downward, ibid. 271, 6159 (1996).
23. D. L. Barber et al., Blood 89, 3166 (1997).
24. M. R. Rozakis-Adcock, R. Fernley, J. Wade, T. Pawson, D. Bowtell, Nature 363, 83 (1993); S. E. Egan et al., ibid, p. 45; N. Li et al., ibid., p. 85; N. W. Gale, S. Kaplan, E. J. Lowenstein, J. Schlessinger, ibid., p. 88; L. Buday and J. Downword, Cell 73, 611 (1993); K. Baltensperger et al., Science 260, 1950 (1993); E. Y. Skolnik et al., ibid., p. 1953.
25. M. R. Rozakis-Adcock, R. Fernley, J. Wade, T. Pawson, D. Bowtell, Nature 363, 83 (1993); S. E. Egan et al., ibid, p. 45; N. Li et al., ibid., p. 85; N. W. Gale, S. Kaplan, E. J. Lowenstein, J. Schlessinger, ibid., p. 88; L. Buday and J. Downword, Cell 73, 611 (1993); K. Baltensperger et al., Science 260, 1950 (1993); E. Y. Skolnik et al., ibid., p. 1953.
26. M. R. Rozakis-Adcock, R. Fernley, J. Wade, T. Pawson, D. Bowtell, Nature 363, 83 (1993); S. E. Egan et al., ibid, p. 45; N. Li et al., ibid., p. 85; N. W. Gale, S. Kaplan, E. J. Lowenstein, J. Schlessinger, ibid., p. 88; L. Buday and J. Downword, Cell 73, 611 (1993); K. Baltensperger et al., Science 260, 1950 (1993); E. Y. Skolnik et al., ibid., p. 1953.
27. M. R. Rozakis-Adcock, R. Fernley, J. Wade, T. Pawson, D. Bowtell, Nature 363, 83 (1993); S. E. Egan et al., ibid, p. 45; N. Li et al., ibid., p. 85; N. W. Gale, S. Kaplan, E. J. Lowenstein, J. Schlessinger, ibid., p. 88; L. Buday and J. Downword, Cell 73, 611 (1993); K. Baltensperger et al., Science 260, 1950 (1993); E. Y. Skolnik et al., ibid., p. 1953.
28. M. R. Rozakis-Adcock, R. Fernley, J. Wade, T. Pawson, D. Bowtell, Nature 363, 83 (1993); S. E. Egan et al., ibid, p. 45; N. Li et al., ibid., p. 85; N. W. Gale, S. Kaplan, E. J. Lowenstein, J. Schlessinger, ibid., p. 88; L. Buday and J. Downword, Cell 73, 611 (1993); K. Baltensperger et al., Science 260, 1950 (1993); E. Y. Skolnik et al., ibid., p. 1953.
29. M. R. Rozakis-Adcock, R. Fernley, J. Wade, T. Pawson, D. Bowtell, Nature 363, 83 (1993); S. E. Egan et al., ibid, p. 45; N. Li et al., ibid., p. 85; N. W. Gale, S. Kaplan, E. J. Lowenstein, J. Schlessinger, ibid., p. 88; L. Buday and J. Downword, Cell 73, 611 (1993); K. Baltensperger et al., Science 260, 1950 (1993); E. Y. Skolnik et al., ibid., p. 1953.
30. M. R. Rozakis-Adcock, R. Fernley, J. Wade, T. Pawson, D. Bowtell, Nature 363, 83 (1993); S. E. Egan et al., ibid, p. 45; N. Li et al., ibid., p. 85; N. W. Gale, S. Kaplan, E. J. Lowenstein, J. Schlessinger, ibid., p. 88; L. Buday and J. Downword, Cell 73, 611 (1993); K. Baltensperger et al., Science 260, 1950 (1993); E. Y. Skolnik et al., ibid., p. 1953.
31. S. Tanaka et al., Proc. Natl. Acad. Sci. U.S.A. 91, 3443 (1994); T. Gotoh et al., Mol. Cell. Biol. 15, 6746 (1995).
32. S. Tanaka et al., Proc. Natl. Acad. Sci. U.S.A. 91, 3443 (1994); T. Gotoh et al., Mol. Cell. Biol. 15, 6746 (1995).
33. S. B. Waters, K. Yamauchi, J. E. Pessin, Mol. Cell. Biol. 15, 2791 (1995); S. B. Waters et al., J. Biol. Chem. 271, 18224 (1996); A. D. Cherniack, J. K. Klarlund, B. R. Conway, M. P. Czech, ibid. 270, 1485 (1995).
34. S. B. Waters, K. Yamauchi, J. E. Pessin, Mol. Cell. Biol. 15, 2791 (1995); S. B. Waters et al., J. Biol. Chem. 271, 18224 (1996); A. D. Cherniack, J. K. Klarlund, B. R. Conway, M. P. Czech, ibid. 270, 1485 (1995).
35. S. B. Waters, K. Yamauchi, J. E. Pessin, Mol. Cell. Biol. 15, 2791 (1995); S. B. Waters et al., J. Biol. Chem. 271, 18224 (1996); A. D. Cherniack, J. K. Klarlund, B. R. Conway, M. P. Czech, ibid. 270, 1485 (1995).
36. V. Pizon, P. Chardin, I. Lerosey, B. Olofsson, A. Tavitian, Oncogene 3, 201 (1988).
37. H. Kitayama, Y. Sugimoto, T. Matsuzaki, Y. Ikawa, M. Noda, Cell 56, 77 (1989).
38. Y. Hata et al., J. Biol. Chem. 265, 7104 (1990).
39. 32P labeling, lysis, immunoprecipitation, elution of guanidine nucleotides bound to the proteins, and calculation of the GTP/(GTP+GDP) ratio were performed as described (30, 36).
40. H. Kitayama, T. Matsuzaki, Y. Ikawa, M. Noda, Proc. Natl. Acad. Sci. U.S.A. 87, 4284 (1990).
41. 5 viable cells were cultured in duplicates with either media or with a combination of TCR mAb [2Ad2, immunoglobulin M (IgM), 1:200] plus CD28 mAb (3D10, IgG2a, 5 μg/ml) or a combination of phorbol 12-myristate 13-acetate (PMA) (50 ng/ml) plus ionomycin (1 μM) for 6 hours. Cell lysates were prepared and measured for luciferase activity as described (38). For titration experiments Jurkat cells were transiently transfected with 20 μg of reporter construct plL-2 luciferase and variable amounts (1.9, 5.6, 16.7, or 50 μg) of pAXEF vector containing Rap1-63E or Rap1-WT cDNA. The total amount of transfected cDNA was normalized to 50 μg by cotransfecting vector cDNA.
42. S. J. Cook, B. Rubinfeld, I. Albert, F. McCormick, EMBO J. 12, 3475 (1993); G. Bokoch, in GTPases in Biology I, B. F. Dickey, and L. Bimbauer, Eds. (Springer, Berlin and Heidelberg, 1993, vol. 108/1), pp. 377-393; I. K. Hariharen, R. W. Carthew, G. M. Rubin, Cell 67, 717 (1991).
43. S. J. Cook, B. Rubinfeld, I. Albert, F. McCormick, EMBO J. 12, 3475 (1993); G. Bokoch, in GTPases in Biology I, B. F. Dickey, and L. Bimbauer, Eds. (Springer, Berlin and Heidelberg, 1993, vol. 108/1), pp. 377-393; I. K. Hariharen, R. W. Carthew, G. M. Rubin, Cell 67, 717 (1991).
44. S. J. Cook, B. Rubinfeld, I. Albert, F. McCormick, EMBO J. 12, 3475 (1993); G. Bokoch, in GTPases in Biology I, B. F. Dickey, and L. Bimbauer, Eds. (Springer, Berlin and Heidelberg, 1993, vol. 108/1), pp. 377-393; I. K. Hariharen, R. W. Carthew, G. M. Rubin, Cell 67, 717 (1991).
45. N. Nassar et al., Nature 375, 554 (1995).
46. M. Frech et al., Science 249, 169 (1990); F. Hofer, S. Fields, C. Schneider, G. S. Martin, Proc. Natl. Acad. Sci. U.S.A. 91, 11089 (1994); A. Kikuchi, S. D. Demo, Z.-H. Ye, Y.-W. Chen, L. T. Williams, Mol. Cell. Biol. 14, 7483 (1994); M. Spaargaren and J. R. Bishoff, Proc. Natl. Acad. Sci. U.S.A. 91, 12609 (1994); F. Beranger, B. Goud, A. Tavitian, J. De-Gunzburg, ibid. 88, 1606 (1991).
47. M. Frech et al., Science 249, 169 (1990); F. Hofer, S. Fields, C. Schneider, G. S. Martin, Proc. Natl. Acad. Sci. U.S.A. 91, 11089 (1994); A. Kikuchi, S. D. Demo, Z.-H. Ye, Y.-W. Chen, L. T. Williams, Mol. Cell. Biol. 14, 7483 (1994); M. Spaargaren and J. R. Bishoff, Proc. Natl. Acad. Sci. U.S.A. 91, 12609 (1994); F. Beranger, B. Goud, A. Tavitian, J. De-Gunzburg, ibid. 88, 1606 (1991).
48. M. Frech et al., Science 249, 169 (1990); F. Hofer, S. Fields, C. Schneider, G. S. Martin, Proc. Natl. Acad. Sci. U.S.A. 91, 11089 (1994); A. Kikuchi, S. D. Demo, Z.-H. Ye, Y.-W. Chen, L. T. Williams, Mol. Cell. Biol. 14, 7483 (1994); M. Spaargaren and J. R. Bishoff, Proc. Natl. Acad. Sci. U.S.A. 91, 12609 (1994); F. Beranger, B. Goud, A. Tavitian, J. De-Gunzburg, ibid. 88, 1606 (1991).
49. M. Frech et al., Science 249, 169 (1990); F. Hofer, S. Fields, C. Schneider, G. S. Martin, Proc. Natl. Acad. Sci. U.S.A. 91, 11089 (1994); A. Kikuchi, S. D. Demo, Z.-H. Ye, Y.-W. Chen, L. T. Williams, Mol. Cell. Biol. 14, 7483 (1994); M. Spaargaren and J. R. Bishoff, Proc. Natl. Acad. Sci. U.S.A. 91, 12609 (1994); F. Beranger, B. Goud, A. Tavitian, J. De-Gunzburg, ibid. 88, 1606 (1991).
50. M. Frech et al., Science 249, 169 (1990); F. Hofer, S. Fields, C. Schneider, G. S. Martin, Proc. Natl. Acad. Sci. U.S.A. 91, 11089 (1994); A. Kikuchi, S. D. Demo, Z.-H. Ye, Y.-W. Chen, L. T. Williams, Mol. Cell. Biol. 14, 7483 (1994); M. Spaargaren and J. R. Bishoff, Proc. Natl. Acad. Sci. U.S.A. 91, 12609 (1994); F. Beranger, B. Goud, A. Tavitian, J. De-Gunzburg, ibid. 88, 1606 (1991).
51. P. H. Warne, P. R. Rodriguez Viciana, J. Downward, Nature 364, 352 (1993); H. Koide, T. Satoh, M. Nakafuku, Y. Kaziro, Proc. Natl. Acad. Sci. U.S.A. 90, 8683 (1993); X. F. Zhang et al., Nature 364, 308 (1993); S. A. Moodie, B. M. Willumsen, M. J. Weber, A. Wolfman, Science 260, 1658 (1993); L. Van Aelst, M. Barr, S. Marcus, A. Polverino, M. Wingler, Proc. Natl. Acad. Sci. U.S.A. 90, 6213 (1993); A. B. Vojtek, S. M. Hollenberg, J. A. Cooper, Cell 74, 205 (1993).
52. P. H. Warne, P. R. Rodriguez Viciana, J. Downward, Nature 364, 352 (1993); H. Koide, T. Satoh, M. Nakafuku, Y. Kaziro, Proc. Natl. Acad. Sci. U.S.A. 90, 8683 (1993); X. F. Zhang et al., Nature 364, 308 (1993); S. A. Moodie, B. M. Willumsen, M. J. Weber, A. Wolfman, Science 260, 1658 (1993); L. Van Aelst, M. Barr, S. Marcus, A. Polverino, M. Wingler, Proc. Natl. Acad. Sci. U.S.A. 90, 6213 (1993); A. B. Vojtek, S. M. Hollenberg, J. A. Cooper, Cell 74, 205 (1993).
53. P. H. Warne, P. R. Rodriguez Viciana, J. Downward, Nature 364, 352 (1993); H. Koide, T. Satoh, M. Nakafuku, Y. Kaziro, Proc. Natl. Acad. Sci. U.S.A. 90, 8683 (1993); X. F. Zhang et al., Nature 364, 308 (1993); S. A. Moodie, B. M. Willumsen, M. J. Weber, A. Wolfman, Science 260, 1658 (1993); L. Van Aelst, M. Barr, S. Marcus, A. Polverino, M. Wingler, Proc. Natl. Acad. Sci. U.S.A. 90, 6213 (1993); A. B. Vojtek, S. M. Hollenberg, J. A. Cooper, Cell 74, 205 (1993).
54. P. H. Warne, P. R. Rodriguez Viciana, J. Downward, Nature 364, 352 (1993); H. Koide, T. Satoh, M. Nakafuku, Y. Kaziro, Proc. Natl. Acad. Sci. U.S.A. 90, 8683 (1993); X. F. Zhang et al., Nature 364, 308 (1993); S. A. Moodie, B. M. Willumsen, M. J. Weber, A. Wolfman, Science 260, 1658 (1993); L. Van Aelst, M. Barr, S. Marcus, A. Polverino, M. Wingler, Proc. Natl. Acad. Sci. U.S.A. 90, 6213 (1993); A. B. Vojtek, S. M. Hollenberg, J. A. Cooper, Cell 74, 205 (1993).
55. P. H. Warne, P. R. Rodriguez Viciana, J. Downward, Nature 364, 352 (1993); H. Koide, T. Satoh, M. Nakafuku, Y. Kaziro, Proc. Natl. Acad. Sci. U.S.A. 90, 8683 (1993); X. F. Zhang et al., Nature 364, 308 (1993); S. A. Moodie, B. M. Willumsen, M. J. Weber, A. Wolfman, Science 260, 1658 (1993); L. Van Aelst, M. Barr, S. Marcus, A. Polverino, M. Wingler, Proc. Natl. Acad. Sci. U.S.A. 90, 6213 (1993); A. B. Vojtek, S. M. Hollenberg, J. A. Cooper, Cell 74, 205 (1993).
56. P. H. Warne, P. R. Rodriguez Viciana, J. Downward, Nature 364, 352 (1993); H. Koide, T. Satoh, M. Nakafuku, Y. Kaziro, Proc. Natl. Acad. Sci. U.S.A. 90, 8683 (1993); X. F. Zhang et al., Nature 364, 308 (1993); S. A. Moodie, B. M. Willumsen, M. J. Weber, A. Wolfman, Science 260, 1658 (1993); L. Van Aelst, M. Barr, S. Marcus, A. Polverino, M. Wingler, Proc. Natl. Acad. Sci. U.S.A. 90, 6213 (1993); A. B. Vojtek, S. M. Hollenberg, J. A. Cooper, Cell 74, 205 (1993).
57. D. Stokoe, S. G. Macdonald, K. Cadwallader, M. Symonsa, J. F. Hancock, Science 264, 1463 (1994); S. J. Leevers, H. J. Paterson, C. J. Marshall, Nature 369, 411 (1994).
58. D. Stokoe, S. G. Macdonald, K. Cadwallader, M. Symonsa, J. F. Hancock, Science 264, 1463 (1994); S. J. Leevers, H. J. Paterson, C. J. Marshall, Nature 369, 411 (1994).
59. C. F. Zheng and K. L Guan, EMBO J. 13, 1123 (1994); D. R. Alessi et al., ibid, p. 1610.
60. C. F. Zheng and K. L Guan, EMBO J. 13, 1123 (1994); D. R. Alessi et al., ibid, p. 1610.
61. J. A. Nunes, Y. Collette, A. Truneh, D. Olive, D. A. Cantrell, J. Exp. Med. 180, 1067 (1994); M. Izquierdo, D. Reif, D. Cantrell, Immunol. Today 16, 159 (1995).
62. J. A. Nunes, Y. Collette, A. Truneh, D. Olive, D. A. Cantrell, J. Exp. Med. 180, 1067 (1994); M. Izquierdo, D. Reif, D. Cantrell, Immunol. Today 16, 159 (1995).
63. M. P. Cooke, K. A. Abraham, K. A. Forbush, R. D. Perlmutter, Cell 65, 281 (1991); N. Glaichenhaus, N. Shastri, D. R. Littman, J. M. Turner, ibid. 64, 511 (1991); S. H. Chan, M. Iwashima, C. W. Turck, A. Weiss, ibid. 71, 649 (1992); R. D. Klausner et al., J. Biol. Chem. 262, 12654 (1987); D. Quian, I. Griswold-Prenner, M. Rieh Rosner, F. W. Fitch, ibid. 268, 4488 (1993); K. S. Ravichandran et al., Science 262, 902 (1993); J. Downward, J. D. Graves, S. Warne, S. Rayter, D. A. Cantrell, Nature 346, 719 (1990); S. M. Rayter, S. C. Woodrow, S. C. Lucas, D. Cantrell, J. Downward, EMBO J. 11, 4549 (1992).
64. M. P. Cooke, K. A. Abraham, K. A. Forbush, R. D. Perlmutter, Cell 65, 281 (1991); N. Glaichenhaus, N. Shastri, D. R. Littman, J. M. Turner, ibid. 64, 511 (1991); S. H. Chan, M. Iwashima, C. W. Turck, A. Weiss, ibid. 71, 649 (1992); R. D. Klausner et al., J. Biol. Chem. 262, 12654 (1987); D. Quian, I. Griswold-Prenner, M. Rieh Rosner, F. W. Fitch, ibid. 268, 4488 (1993); K. S. Ravichandran et al., Science 262, 902 (1993); J. Downward, J. D. Graves, S. Warne, S. Rayter, D. A. Cantrell, Nature 346, 719 (1990); S. M. Rayter, S. C. Woodrow, S. C. Lucas, D. Cantrell, J. Downward, EMBO J. 11, 4549 (1992).
65. M. P. Cooke, K. A. Abraham, K. A. Forbush, R. D. Perlmutter, Cell 65, 281 (1991); N. Glaichenhaus, N. Shastri, D. R. Littman, J. M. Turner, ibid. 64, 511 (1991); S. H. Chan, M. Iwashima, C. W. Turck, A. Weiss, ibid. 71, 649 (1992); R. D. Klausner et al., J. Biol. Chem. 262, 12654 (1987); D. Quian, I. Griswold-Prenner, M. Rieh Rosner, F. W. Fitch, ibid. 268, 4488 (1993); K. S. Ravichandran et al., Science 262, 902 (1993); J. Downward, J. D. Graves, S. Warne, S. Rayter, D. A. Cantrell, Nature 346, 719 (1990); S. M. Rayter, S. C. Woodrow, S. C. Lucas, D. Cantrell, J. Downward, EMBO J. 11, 4549 (1992).
66. M. P. Cooke, K. A. Abraham, K. A. Forbush, R. D. Perlmutter, Cell 65, 281 (1991); N. Glaichenhaus, N. Shastri, D. R. Littman, J. M. Turner, ibid. 64, 511 (1991); S. H. Chan, M. Iwashima, C. W. Turck, A. Weiss, ibid. 71, 649 (1992); R. D. Klausner et al., J. Biol. Chem. 262, 12654 (1987); D. Quian, I. Griswold-Prenner, M. Rieh Rosner, F. W. Fitch, ibid. 268, 4488 (1993); K. S. Ravichandran et al., Science 262, 902 (1993); J. Downward, J. D. Graves, S. Warne, S. Rayter, D. A. Cantrell, Nature 346, 719 (1990); S. M. Rayter, S. C. Woodrow, S. C. Lucas, D. Cantrell, J. Downward, EMBO J. 11, 4549 (1992).
67. M. P. Cooke, K. A. Abraham, K. A. Forbush, R. D. Perlmutter, Cell 65, 281 (1991); N. Glaichenhaus, N. Shastri, D. R. Littman, J. M. Turner, ibid. 64, 511 (1991); S. H. Chan, M. Iwashima, C. W. Turck, A. Weiss, ibid. 71, 649 (1992); R. D. Klausner et al., J. Biol. Chem. 262, 12654 (1987); D. Quian, I. Griswold-Prenner, M. Rieh Rosner, F. W. Fitch, ibid. 268, 4488 (1993); K. S. Ravichandran et al., Science 262, 902 (1993); J. Downward, J. D. Graves, S. Warne, S. Rayter, D. A. Cantrell, Nature 346, 719 (1990); S. M. Rayter, S. C. Woodrow, S. C. Lucas, D. Cantrell, J. Downward, EMBO J. 11, 4549 (1992).
68. M. P. Cooke, K. A. Abraham, K. A. Forbush, R. D. Perlmutter, Cell 65, 281 (1991); N. Glaichenhaus, N. Shastri, D. R. Littman, J. M. Turner, ibid. 64, 511 (1991); S. H. Chan, M. Iwashima, C. W. Turck, A. Weiss, ibid. 71, 649 (1992); R. D. Klausner et al., J. Biol. Chem. 262, 12654 (1987); D. Quian, I. Griswold-Prenner, M. Rieh Rosner, F. W. Fitch, ibid. 268, 4488 (1993); K. S. Ravichandran et al., Science 262, 902 (1993); J. Downward, J. D. Graves, S. Warne, S. Rayter, D. A. Cantrell, Nature 346, 719 (1990); S. M. Rayter, S. C. Woodrow, S. C. Lucas, D. Cantrell, J. Downward, EMBO J. 11, 4549 (1992).
69. M. P. Cooke, K. A. Abraham, K. A. Forbush, R. D. Perlmutter, Cell 65, 281 (1991); N. Glaichenhaus, N. Shastri, D. R. Littman, J. M. Turner, ibid. 64, 511 (1991); S. H. Chan, M. Iwashima, C. W. Turck, A. Weiss, ibid. 71, 649 (1992); R. D. Klausner et al., J. Biol. Chem. 262, 12654 (1987); D. Quian, I. Griswold-Prenner, M. Rieh Rosner, F. W. Fitch, ibid. 268, 4488 (1993); K. S. Ravichandran et al., Science 262, 902 (1993); J. Downward, J. D. Graves, S. Warne, S. Rayter, D. A. Cantrell, Nature 346, 719 (1990); S. M. Rayter, S. C. Woodrow, S. C. Lucas, D. Cantrell, J. Downward, EMBO J. 11, 4549 (1992).
70. M. P. Cooke, K. A. Abraham, K. A. Forbush, R. D. Perlmutter, Cell 65, 281 (1991); N. Glaichenhaus, N. Shastri, D. R. Littman, J. M. Turner, ibid. 64, 511 (1991); S. H. Chan, M. Iwashima, C. W. Turck, A. Weiss, ibid. 71, 649 (1992); R. D. Klausner et al., J. Biol. Chem. 262, 12654 (1987); D. Quian, I. Griswold-Prenner, M. Rieh Rosner, F. W. Fitch, ibid. 268, 4488 (1993); K. S. Ravichandran et al., Science 262, 902 (1993); J. Downward, J. D. Graves, S. Warne, S. Rayter, D. A. Cantrell, Nature 346, 719 (1990); S. M. Rayter, S. C. Woodrow, S. C. Lucas, D. Cantrell, J. Downward, EMBO J. 11, 4549 (1992).
71. C. H. Yoon, J. Lee, G. D. Jongeward, P. W. Sternberg, Science 269, 1102 (1995); G. D. Jongeward, T. R. Clandinin, P. W. Sternberg, Genetics 139, 1553 (1995).
72. C. H. Yoon, J. Lee, G. D. Jongeward, P. W. Sternberg, Science 269, 1102 (1995); G. D. Jongeward, T. R. Clandinin, P. W. Sternberg, Genetics 139, 1553 (1995).
73. 7 cells/ml and incubated with anti-CD3 (OKT3, IgG1, American Type Culture Collection) (1 μg/ml) and anti-CD28 (3D10, IgG2a) (5 μg/ml) at 4°C for 20 min. Cells were washed and incubated with rabbit antibody to mouse Ig (10 μg/ml) at 37°C for 1 min that could induce maximum activation and phosphorylation of Fyn in control cells (11).
74. K. Xia et al., Proc. Natl. Acad. Sci. U.S.A. 93, 11681 (1996); D. L. Barber, C. N. Corless, K. Xia, T. Roberts, A. D. D'Andrea, Blood 89, 55 (1997).
75. K. Xia et al., Proc. Natl. Acad. Sci. U.S.A. 93, 11681 (1996); D. L. Barber, C. N. Corless, K. Xia, T. Roberts, A. D. D'Andrea, Blood 89, 55 (1997).
76. B. Hallberg, S. I. Rayter, J. Downward, J. Biol. Chem. 269, 3913 (1994).
77. H. L. Brownell et al., DNA Cell Biol. 15, 443 (1996).
78. N. A. Clipstone and G. R. Crabtree, Nature 357, 695 (1992).
79. H. Pahl, T. Burn, D. Tenen, Exp. Hematol. 19, 1038 (1991); D. R. Plas et al., Science 272, 1173 (1996); D. G. Motto, S. E. Ross, J. Wu, L. R. Hendricks-Taylor, G. A. Koretzky, J. Exp. Med. 183, 1937 (1996).
80. H. Pahl, T. Burn, D. Tenen, Exp. Hematol. 19, 1038 (1991); D. R. Plas et al., Science 272, 1173 (1996); D. G. Motto, S. E. Ross, J. Wu, L. R. Hendricks-Taylor, G. A. Koretzky, J. Exp. Med. 183, 1937 (1996).
81. H. Pahl, T. Burn, D. Tenen, Exp. Hematol. 19, 1038 (1991); D. R. Plas et al., Science 272, 1173 (1996); D. G. Motto, S. E. Ross, J. Wu, L. R. Hendricks-Taylor, G. A. Koretzky, J. Exp. Med. 183, 1937 (1996).
82. We thank H. Kitayama and M. Noda for providing the Rap-1 (Krev) and Rap1-63E plasmids, D. McKean and R. Abraham for providing the IL-2-luciferase plasmid, T. Roberts and R. Narsimhan for helpful suggestions on the GTP and GDP binding assays, D. Tenen for assistance with the luciferase assays, and N. Malencovitch for technical support. Supported by NIH grants Al 35225, HL 54785, and Al39671. D.L.B. is a Special Fellow of the Leukemia Society of America.