Fidelity of T cell activation through multistep T cell receptor ζ phosphorylation

Science

Volumn 281, Issue 5376, 1998, Pages 572-575

  Kersh, Ellen Neumeister ^a   Shaw, Andrey S ^a   Allen, Paul M ^a  

^a WASHINGTON UNIVERSITY SCHOOL OF MEDICINE   (United States)

Author keywords

[No Author keywords available]

Indexed keywords

MONOCLONAL ANTIBODY; PHOSPHOTYROSINE; T LYMPHOCYTE RECEPTOR;

ANIMAL CELL; ANTIGEN RECOGNITION; ARTICLE; BINDING AFFINITY; LIGAND BINDING; MOUSE; NONHUMAN; PRIORITY JOURNAL; PROTEIN PHOSPHORYLATION; RECEPTOR BINDING; T LYMPHOCYTE ACTIVATION;

ANIMALS; ANTIGEN-PRESENTING CELLS; ANTIGENS; ANTIGENS, CD3; LIGANDS; LYMPHOCYTE ACTIVATION; MEMBRANE PROTEINS; MICE; MICE, TRANSGENIC; MUTATION; PEPTIDES; PHOSPHORYLATION; PHOSPHOTYROSINE; PROTEIN-TYROSINE KINASES; RECEPTORS, ANTIGEN, T-CELL; RECEPTORS, ANTIGEN, T-CELL, ALPHA-BETA; SIGNAL TRANSDUCTION; T-LYMPHOCYTES; ZAP-70 PROTEIN-TYROSINE KINASE;

ANIMALIA; MURINAE;

EID: 0032563231     PISSN: 00368075     EISSN: None     Source Type: Journal    
DOI: 10.1126/science.281.5376.572     Document Type: Article

References (74)

1. L. E. Samelson, J. B. Harford, R. D. Klausner, Cell 43, 223 (1985); M. Reth, Nature 338, 383 (1989), A. Weiss and D. R. Littman, Cell 76, 263 (1994); A. C. Chan and A. S. Shaw, Curr. Opin. Immunol. 8, 394 (1996); R. L. Wange and L. E. Samelson, Immunity 5, 197 (1996).
2. L. E. Samelson, J. B. Harford, R. D. Klausner, Cell 43, 223 (1985); M. Reth, Nature 338, 383 (1989), A. Weiss and D. R. Littman, Cell 76, 263 (1994); A. C. Chan and A. S. Shaw, Curr. Opin. Immunol. 8, 394 (1996); R. L. Wange and L. E. Samelson, Immunity 5, 197 (1996).
3. L. E. Samelson, J. B. Harford, R. D. Klausner, Cell 43, 223 (1985); M. Reth, Nature 338, 383 (1989), A. Weiss and D. R. Littman, Cell 76, 263 (1994); A. C. Chan and A. S. Shaw, Curr. Opin. Immunol. 8, 394 (1996); R. L. Wange and L. E. Samelson, Immunity 5, 197 (1996).
4. L. E. Samelson, J. B. Harford, R. D. Klausner, Cell 43, 223 (1985); M. Reth, Nature 338, 383 (1989), A. Weiss and D. R. Littman, Cell 76, 263 (1994); A. C. Chan and A. S. Shaw, Curr. Opin. Immunol. 8, 394 (1996); R. L. Wange and L. E. Samelson, Immunity 5, 197 (1996).
5. L. E. Samelson, J. B. Harford, R. D. Klausner, Cell 43, 223 (1985); M. Reth, Nature 338, 383 (1989), A. Weiss and D. R. Littman, Cell 76, 263 (1994); A. C. Chan and A. S. Shaw, Curr. Opin. Immunol. 8, 394 (1996); R. L. Wange and L. E. Samelson, Immunity 5, 197 (1996).
6. Single-letter 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. X indicates any residue.
7. S. J. Frank et al., Science 249, 174 (1990); B. A. Irving and A. Weiss, Cell 64, 891 (1991); C. Romeo, M. Amiot B. Seed, ibid. 68, 889 (1992); F. Letourneur and R. D. Klausner, Science 255, 79 (1992); B. A. Irving, A. C. Chan, A. Weiss, J. Exp. Med. 177, 1093 (1993), F. Letourneur and R. D. Klausner, Proc. Natl. Acad. Sci. U.S.A. 88, 8905 (1991).
8. S. J. Frank et al., Science 249, 174 (1990); B. A. Irving and A. Weiss, Cell 64, 891 (1991); C. Romeo, M. Amiot B. Seed, ibid. 68, 889 (1992); F. Letourneur and R. D. Klausner, Science 255, 79 (1992); B. A. Irving, A. C. Chan, A. Weiss, J. Exp. Med. 177, 1093 (1993), F. Letourneur and R. D. Klausner, Proc. Natl. Acad. Sci. U.S.A. 88, 8905 (1991).
9. S. J. Frank et al., Science 249, 174 (1990); B. A. Irving and A. Weiss, Cell 64, 891 (1991); C. Romeo, M. Amiot B. Seed, ibid. 68, 889 (1992); F. Letourneur and R. D. Klausner, Science 255, 79 (1992); B. A. Irving, A. C. Chan, A. Weiss, J. Exp. Med. 177, 1093 (1993), F. Letourneur and R. D. Klausner, Proc. Natl. Acad. Sci. U.S.A. 88, 8905 (1991).
10. S. J. Frank et al., Science 249, 174 (1990); B. A. Irving and A. Weiss, Cell 64, 891 (1991); C. Romeo, M. Amiot B. Seed, ibid. 68, 889 (1992); F. Letourneur and R. D. Klausner, Science 255, 79 (1992); B. A. Irving, A. C. Chan, A. Weiss, J. Exp. Med. 177, 1093 (1993), F. Letourneur and R. D. Klausner, Proc. Natl. Acad. Sci. U.S.A. 88, 8905 (1991).
11. S. J. Frank et al., Science 249, 174 (1990); B. A. Irving and A. Weiss, Cell 64, 891 (1991); C. Romeo, M. Amiot B. Seed, ibid. 68, 889 (1992); F. Letourneur and R. D. Klausner, Science 255, 79 (1992); B. A. Irving, A. C. Chan, A. Weiss, J. Exp. Med. 177, 1093 (1993), F. Letourneur and R. D. Klausner, Proc. Natl. Acad. Sci. U.S.A. 88, 8905 (1991).
12. S. J. Frank et al., Science 249, 174 (1990); B. A. Irving and A. Weiss, Cell 64, 891 (1991); C. Romeo, M. Amiot B. Seed, ibid. 68, 889 (1992); F. Letourneur and R. D. Klausner, Science 255, 79 (1992); B. A. Irving, A. C. Chan, A. Weiss, J. Exp. Med. 177, 1093 (1993), F. Letourneur and R. D. Klausner, Proc. Natl. Acad. Sci. U.S.A. 88, 8905 (1991).
13. E. W. Shores and P. E. Love, Curr. Opin. Immunol. 9, 380 (1997); P. E. Love et al., Science 261, 918 (1993); C.-P. Liu et al., EMBO J. 12, 4863 (1993); M. A. Malissen et al., ibid., p. 4347; H. Ohno et al., ibid., p. 4357.
14. E. W. Shores and P. E. Love, Curr. Opin. Immunol. 9, 380 (1997); P. E. Love et al., Science 261, 918 (1993); C.-P. Liu et al., EMBO J. 12, 4863 (1993); M. A. Malissen et al., ibid., p. 4347; H. Ohno et al., ibid., p. 4357.
15. E. W. Shores and P. E. Love, Curr. Opin. Immunol. 9, 380 (1997); P. E. Love et al., Science 261, 918 (1993); C.-P. Liu et al., EMBO J. 12, 4863 (1993); M. A. Malissen et al., ibid., p. 4347; H. Ohno et al., ibid., p. 4357.
16. E. W. Shores and P. E. Love, Curr. Opin. Immunol. 9, 380 (1997); P. E. Love et al., Science 261, 918 (1993); C.-P. Liu et al., EMBO J. 12, 4863 (1993); M. A. Malissen et al., ibid., p. 4347; H. Ohno et al., ibid., p. 4357.
17. E. W. Shores and P. E. Love, Curr. Opin. Immunol. 9, 380 (1997); P. E. Love et al., Science 261, 918 (1993); C.-P. Liu et al., EMBO J. 12, 4863 (1993); M. A. Malissen et al., ibid., p. 4347; H. Ohno et al., ibid., p. 4357.
18. E. W. Shores et al., Science 266, 1047 (1994); C.-P. Liu, W.-J. Lin, M. Huang, J. W. Kappler, P. Marrack, Proc. Natl. Acad. Sci. U.S.A. 94, 616 (1997); E. Shores et al., J. Immunol. 159, 222 (1997).
19. E. W. Shores et al., Science 266, 1047 (1994); C.-P. Liu, W.-J. Lin, M. Huang, J. W. Kappler, P. Marrack, Proc. Natl. Acad. Sci. U.S.A. 94, 616 (1997); E. Shores et al., J. Immunol. 159, 222 (1997).
20. E. W. Shores et al., Science 266, 1047 (1994); C.-P. Liu, W.-J. Lin, M. Huang, J. W. Kappler, P. Marrack, Proc. Natl. Acad. Sci. U.S.A. 94, 616 (1997); E. Shores et al., J. Immunol. 159, 222 (1997).
21. E. W. Shores et al., J. Exp. Med. 185, 893 (1997); T. Yamazaki et al., J. Immunol. 158, 1634 (1997); S.-Y. Lin, L. Ardouin, A. Gillet, M. Malissen, B. Malissen, J. Exp. Med. 185, 707 (1997).
22. E. W. Shores et al., J. Exp. Med. 185, 893 (1997); T. Yamazaki et al., J. Immunol. 158, 1634 (1997); S.-Y. Lin, L. Ardouin, A. Gillet, M. Malissen, B. Malissen, J. Exp. Med. 185, 707 (1997).
23. E. W. Shores et al., J. Exp. Med. 185, 893 (1997); T. Yamazaki et al., J. Immunol. 158, 1634 (1997); S.-Y. Lin, L. Ardouin, A. Gillet, M. Malissen, B. Malissen, J. Exp. Med. 185, 707 (1997).
24. D. Qian, I. Griswold-Prenner, M. R. Rosner, F. W. Fitch, J. Biol. Chem. 268, 4488 (1993); T. Nakayama, A. Singer, E. D. Hsi, L E. Samelson, Nature 341, 651 (1989); C. Cenciarelli et al., Science 257, 795 (1992); N. S. C. van Oers et al., Mol. Cell. Biol. 13, 5771 (1993).
25. D. Qian, I. Griswold-Prenner, M. R. Rosner, F. W. Fitch, J. Biol. Chem. 268, 4488 (1993); T. Nakayama, A. Singer, E. D. Hsi, L E. Samelson, Nature 341, 651 (1989); C. Cenciarelli et al., Science 257, 795 (1992); N. S. C. van Oers et al., Mol. Cell. Biol. 13, 5771 (1993).
26. D. Qian, I. Griswold-Prenner, M. R. Rosner, F. W. Fitch, J. Biol. Chem. 268, 4488 (1993); T. Nakayama, A. Singer, E. D. Hsi, L E. Samelson, Nature 341, 651 (1989); C. Cenciarelli et al., Science 257, 795 (1992); N. S. C. van Oers et al., Mol. Cell. Biol. 13, 5771 (1993).
27. D. Qian, I. Griswold-Prenner, M. R. Rosner, F. W. Fitch, J. Biol. Chem. 268, 4488 (1993); T. Nakayama, A. Singer, E. D. Hsi, L E. Samelson, Nature 341, 651 (1989); C. Cenciarelli et al., Science 257, 795 (1992); N. S. C. van Oers et al., Mol. Cell. Biol. 13, 5771 (1993).
28. N. S. C. van Oers, N. Killeen, A. Weiss, Immunity 1, 675 (1994); D. L. Wiest, J. M. Ashe, R. Abe, J. B. Bolen, A. Singer, ibid. 4, 495 (1996).
29. N. S. C. van Oers, N. Killeen, A. Weiss, Immunity 1, 675 (1994); D. L. Wiest, J. M. Ashe, R. Abe, J. B. Bolen, A. Singer, ibid. 4, 495 (1996).
30. Standard techniques used in the examination of phosphoproteins, such as tryptic mapping or mass spectrometry, have failed to identify specific phospho-species of ζ in resting or activated T cells. Naming of the phospho-isoforms of ζ has been inconsistent in the literature. We will refer to the phosphoform seen in resting T cells as p21, and the additional form in activated T cells as p23.
31. J. Sloan-Lancaster, A. S. Shaw, J. B. Rothbard, P. M. Allen, Cell 79, 913 (1994).
32. J. Madrenas et al., Science 267, 515 (1995); C. Reis e Sousa, E. H. Levine, R. N. Germain, J. Exp. Med. 184, 149 (1996); D. M. La Face et al., J. Immunol. 158, 2057 (1997); J. A. Smith, J. Y. Tso, M. R. Clark, M. S. Cole, J. A. Bluestone, J. Exp. Med. 185, 1413 (1997).
33. J. Madrenas et al., Science 267, 515 (1995); C. Reis e Sousa, E. H. Levine, R. N. Germain, J. Exp. Med. 184, 149 (1996); D. M. La Face et al., J. Immunol. 158, 2057 (1997); J. A. Smith, J. Y. Tso, M. R. Clark, M. S. Cole, J. A. Bluestone, J. Exp. Med. 185, 1413 (1997).
34. J. Madrenas et al., Science 267, 515 (1995); C. Reis e Sousa, E. H. Levine, R. N. Germain, J. Exp. Med. 184, 149 (1996); D. M. La Face et al., J. Immunol. 158, 2057 (1997); J. A. Smith, J. Y. Tso, M. R. Clark, M. S. Cole, J. A. Bluestone, J. Exp. Med. 185, 1413 (1997).
35. J. Madrenas et al., Science 267, 515 (1995); C. Reis e Sousa, E. H. Levine, R. N. Germain, J. Exp. Med. 184, 149 (1996); D. M. La Face et al., J. Immunol. 158, 2057 (1997); J. A. Smith, J. Y. Tso, M. R. Clark, M. S. Cole, J. A. Bluestone, J. Exp. Med. 185, 1413 (1997).
36. -9 M to the protein immunoblotting solution.
37. 4)KGLGK or the unphosphorylated AP1 peptide. Anti-pAZ was purified similarly. Anti-pC2 was purified on the nonspecific phosphotyrosine column only. Anti-pB1, anti-pB2, and anti-pC2 were specific without purification; all sera were titrated to give the best background-to-signal ratio in protein immunoblotting. Antiserum to ζ for protein immunoblotting was raised as in (35), and the mAb to pY was 4G10 (UBI). Protein immunoblots were detected by ECL (Amersham), with techniques according to the manufacturer's instructions, except that blots were washed in 0.5% NP-40 containing phosphate-buffered saline for an additional 15 min after each antibody incubation.
38. B. D. Evavold, S. C. Williams, B. L. Hsu, S. Buus, P. M. Alien, J. Immunol. 148, 347 (1992). The sequence of Hb(64-76) is GKKVITAFNEGLK.
39. 7) in a volume of 250 μl. The cell suspension was incubated at 37°C for the indicated time. Cells were then lysed (10) and precipitated with 10 μg of 500.A2 (an antibody to CD3ε) (Pharmingen) or with anti-ZAP-70. Samples were separated on a 13% SDS-PAGE gel.
40. A2 phosphorylation in resting T cells was observed in four of seven experiments. When it was observed, it was consistently weaker than B1 and C2 phosphorylation when compared to maximal phosphorylation upon T cell activation. Thus, A2 phosphorylation may not be as prominent as B1 and C2 phosphorylation in resting T cells. Anti-pA1 did not recognize p21, but consistently reacted with minor migration forms of p-ζ of less than 21 kD. These minor forms were also present in the protein immunoblot to pY after longer exposure of the film. Anti-pC2 recognized p21; thus, p21 is not a form of η, an alternatively spliced form of ζ that does not contain tyrosine C2. p21 and p23 were not recognized by monoclonal or polyclonal antibodies to ζ, presumably because the amount of protein in p21 and p23 was too low to be detected (17).
41. E. N. Kersh and P. M. Allen, unpublished results.
42. A. Crakoui and P. Alien, in preparation.
43. -/- mice twice with cold Hank's balanced salt solution and then lysed the cells. The TCR complex was precipitated with 10 μg of mAb 500.A2 to CD3ε and analyzed using protein immunoblotting as described for Fig. 2 and 3.
44. Anti-pB2 serum could also recognize p21 in activated T cells, although this was only apparent after long exposure of the film. p21 with pB2 may represent a minor fraction of pζ in the transition to full phosphorylation. Because B2 phosphorylation in p21 of resting T cells was not detected, p21 can probably contain multiple phospho-species that are not resolved electrophoretically. Similarly, p23 contained several species (Fig. 3 and text).
45. Y. Sykulev et al., Immunity 1, 15 (1994); S. M. Alam et al., Nature 381, 616 (1996); D. S. Lyons et al., Immunity 5, 53 (1996).
46. Y. Sykulev et al., Immunity 1, 15 (1994); S. M. Alam et al., Nature 381, 616 (1996); D. S. Lyons et al., Immunity 5, 53 (1996).
47. Y. Sykulev et al., Immunity 1, 15 (1994); S. M. Alam et al., Nature 381, 616 (1996); D. S. Lyons et al., Immunity 5, 53 (1996).
48. J. Sloan-Lancaster and P. M. Allen, Annu. Rev. Immunol. 14, 1 (1996); S. C. Jameson and M. J. Bevan, Immunity 2, 1 (1995); A. Sette et al., Annu. Rev. Immunol. 12, 413 (1994); R. N. Germain, E. H. Levine, J. Madrenas, Immunologist 3, 113 (1995).
49. J. Sloan-Lancaster and P. M. Allen, Annu. Rev. Immunol. 14, 1 (1996); S. C. Jameson and M. J. Bevan, Immunity 2, 1 (1995); A. Sette et al., Annu. Rev. Immunol. 12, 413 (1994); R. N. Germain, E. H. Levine, J. Madrenas, Immunologist 3, 113 (1995).
50. J. Sloan-Lancaster and P. M. Allen, Annu. Rev. Immunol. 14, 1 (1996); S. C. Jameson and M. J. Bevan, Immunity 2, 1 (1995); A. Sette et al., Annu. Rev. Immunol. 12, 413 (1994); R. N. Germain, E. H. Levine, J. Madrenas, Immunologist 3, 113 (1995).
51. J. Sloan-Lancaster and P. M. Allen, Annu. Rev. Immunol. 14, 1 (1996); S. C. Jameson and M. J. Bevan, Immunity 2, 1 (1995); A. Sette et al., Annu. Rev. Immunol. 12, 413 (1994); R. N. Germain, E. H. Levine, J. Madrenas, Immunologist 3, 113 (1995).
52. C. J. Kersh and P. M. Allen J. Exp. Med. 184, 1259 (1996) The amino acid sequences are G72: GKKVITAFGEGLK; 172: GKKVITAFIEGLK; D73; GKKVITAFNDGLK.
53. ZAP-70 activation was detected at reduced amounts after D73 stimulation, when a more sensitive in vitro kinase assay was used (17).
54. T. W. McKeithan, Proc. Natl. Acad. Sci. U.S.A. 92, 5042 (1995); J. D. Rabinowitz, C. Beeson, D. S. Lyons, M. M. Davis, H. M. McConnell, ibid. 93, 1401 (1996).
55. T. W. McKeithan, Proc. Natl. Acad. Sci. U.S.A. 92, 5042 (1995); J. D. Rabinowitz, C. Beeson, D. S. Lyons, M. M. Davis, H. M. McConnell, ibid. 93, 1401 (1996).
56. A.-M. K. Wegener et al., Cell 68, 83 (1992); Y. Shinkai, A. Ma, H.-L. Cheng, F. W. Alt, Immunity 2, 401 (1995).
57. A.-M. K. Wegener et al., Cell 68, 83 (1992); Y. Shinkai, A. Ma, H.-L. Cheng, F. W. Alt, Immunity 2, 401 (1995).
58. N. S. C. van Oers, P. E. Love, E. W. Shores, A. Weiss, J. Immunol. 160, 163 (1998).
59. N. Isakov et al., J. Exp. Med. 181, 375 (1995); J. -Y. Bu, A. S. Shaw, A. C. Chan, Proc. Natl. Acad. Sci. U.S.A. 92, 5106 (1995); F. Vely, J. A. Nunes, B. Malissen, C. J. Hedgecock, Eur. J. Immunol. 27, 3010 (1997).
60. N. Isakov et al., J. Exp. Med. 181, 375 (1995); J. -Y. Bu, A. S. Shaw, A. C. Chan, Proc. Natl. Acad. Sci. U.S.A. 92, 5106 (1995); F. Vely, J. A. Nunes, B. Malissen, C. J. Hedgecock, Eur. J. Immunol. 27, 3010 (1997).
61. N. Isakov et al., J. Exp. Med. 181, 375 (1995); J. -Y. Bu, A. S. Shaw, A. C. Chan, Proc. Natl. Acad. Sci. U.S.A. 92, 5106 (1995); F. Vely, J. A. Nunes, B. Malissen, C. J. Hedgecock, Eur. J. Immunol. 27, 3010 (1997).
62. B. Combadière et al., J. Exp. Med. 183, 2109 (1996).
63. S. Koyasu et al., J. Biol. Chem. 267, 3375 (1992); S. J. Frank et al., ibid., p. 13656.
64. S. Koyasu et al., J. Biol. Chem. 267, 3375 (1992); S. J. Frank et al., ibid., p. 13656.
65. N. S. C. van Oers, N. Killeen, A. Weiss, J. Exp. Med. 183, 1053 (1996); N. S. C. van Oers, B. Lowin-Kropf, D. Finlay, K. Connolly, A. Weiss, Immunity 5, 429 (1996); J. D. Stone et al., J. Immunol. 158, 5773 (1997); D. R. Plas et al., Science 272, 1173 (1996).
66. N. S. C. van Oers, N. Killeen, A. Weiss, J. Exp. Med. 183, 1053 (1996); N. S. C. van Oers, B. Lowin-Kropf, D. Finlay, K. Connolly, A. Weiss, Immunity 5, 429 (1996); J. D. Stone et al., J. Immunol. 158, 5773 (1997); D. R. Plas et al., Science 272, 1173 (1996).
67. N. S. C. van Oers, N. Killeen, A. Weiss, J. Exp. Med. 183, 1053 (1996); N. S. C. van Oers, B. Lowin-Kropf, D. Finlay, K. Connolly, A. Weiss, Immunity 5, 429 (1996); J. D. Stone et al., J. Immunol. 158, 5773 (1997); D. R. Plas et al., Science 272, 1173 (1996).
68. N. S. C. van Oers, N. Killeen, A. Weiss, J. Exp. Med. 183, 1053 (1996); N. S. C. van Oers, B. Lowin-Kropf, D. Finlay, K. Connolly, A. Weiss, Immunity 5, 429 (1996); J. D. Stone et al., J. Immunol. 158, 5773 (1997); D. R. Plas et al., Science 272, 1173 (1996).
69. N. Osman, H. Turner, S. Lucas, K. Reif, D. A. Cantrell, Eur. J. Immunol. 26, 1063 (1996); W. A. Jensen et al., ibid. 27, 707 (1997).
70. N. Osman, H. Turner, S. Lucas, K. Reif, D. A. Cantrell, Eur. J. Immunol. 26, 1063 (1996); W. A. Jensen et al., ibid. 27, 707 (1997).
71. A. M. Weissman et al., Science 239, 1018 (1988).
72. L. K. T. Gauen, A.-N. Tony Kong, L. E. Samelson, A. S. Shaw, Mol. Cell. Biol. 12, 5438 (1992).
73. D. G. Orloff, S. J. Frank, F. A. Robey, A. M. Weissman, R. D. Klausner, J. Biol. Chem. 264, 14812 (1989).
74. -/- mice, A. Chan for the gift of anti-ZAP-70, R. Jordan and S. Horvath for excellent technical assistance, and J. Smith for excellent secretarial assistance. Supported by NIH.