Skip to main content
PMC home page
The EMBO Journal logoLink to The EMBO Journal
. 1995 Jul 17;14(14):3385–3394. doi: 10.1002/j.1460-2075.1995.tb07344.x

Reconstitution of interactions between tyrosine kinases and the high affinity IgE receptor which are controlled by receptor clustering.

A M Scharenberg 1, S Lin 1, B Cuenod 1, H Yamamura 1, J P Kinet 1
PMCID: PMC394405  PMID: 7628439

Abstract

High affinity IgE receptor (Fc epsilon RI) signaling after contact with antigen occurs in response to receptor clustering. This paper describes methodology, based on vaccinia virus driven protein expression, for probing signaling pathways and its application to Fc epsilon RI interactions with the lyn and syk tyrosine kinases. Reconstitution of the complete tetrameric Fc epsilon RI receptor, lyn and syk in a non-hematopoietic 'null' cell line is sufficient to reconstruct clustering-controlled receptor tyrosine phosphorylation and activation of syk, without apparent requirement for hematopoietic specific phosphatases. The src family kinase lyn phosphorylates Fc epsilon RI in response to receptor clustering, resulting in syk binding to the phosphorylated Fc epsilon RI. Lyn also participates in the tyrosine phosphorylation and activation of syk in a manner which is dependent on phosphorylated Fc epsilon RI. Using overexpression of active and dominant negative syk proteins in a mast cell line which naturally expresses Fc epsilon RI, we corroborate syk's role downstream of receptor phosphorylation, and demonstrate that syk SH2 domains protect receptor ITAMs from ongoing dephosphorylation. Based on these results, we propose that receptor clustering controls lyn-mediated Fc epsilon RI tyrosine phosphorylation by shifting a balance between phosphorylation and dephosphorylation towards accumulation of tyrosine phosphorylated Fc epsilon RI. Fc epsilon RI tyrosine phosphorylation functions to bring syk into a microenvironment where it becomes tyrosine phosphorylated and activated, thereby allowing clustering to indirectly control syk activity.

Full text

PDF
3391

Images in this article

3386Image
on p.3386
3387Image
on p.3387
3387Image
on p.3387
3388Image
on p.3388
3389Image
on p.3389
3389Image
on p.3389
3390Image
on p.3390
3390Image
on p.3390

Selected References

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

  1. Adamczewski M., Kinet J. P. The high-affinity receptor for immunoglobulin E. Chem Immunol. 1994;59:173–190. [PubMed] [Google Scholar]
  2. Adamczewski M., Numerof R. P., Koretzky G. A., Kinet J. P. Regulation by CD45 of the tyrosine phosphorylation of high affinity IgE receptor beta- and gamma-chains. J Immunol. 1995 Apr 1;154(7):3047–3055. [PubMed] [Google Scholar]
  3. Beaven M. A., Metzger H. Signal transduction by Fc receptors: the Fc epsilon RI case. Immunol Today. 1993 May;14(5):222–226. doi: 10.1016/0167-5699(93)90167-j. [DOI] [PubMed] [Google Scholar]
  4. Benhamou M., Ryba N. J., Kihara H., Nishikata H., Siraganian R. P. Protein-tyrosine kinase p72syk in high affinity IgE receptor signaling. Identification as a component of pp72 and association with the receptor gamma chain after receptor aggregation. J Biol Chem. 1993 Nov 5;268(31):23318–23324. [PubMed] [Google Scholar]
  5. Benhamou M., Stephan V., Robbins K. C., Siraganian R. P. High-affinity IgE receptor-mediated stimulation of rat basophilic leukemia (RBL-2H3) cells induces early and late protein-tyrosine phosphorylations. J Biol Chem. 1992 Apr 15;267(11):7310–7314. [PubMed] [Google Scholar]
  6. Blank U., Ra C., Miller L., White K., Metzger H., Kinet J. P. Complete structure and expression in transfected cells of high affinity IgE receptor. Nature. 1989 Jan 12;337(6203):187–189. doi: 10.1038/337187a0. [DOI] [PubMed] [Google Scholar]
  7. Bolen J. B., Thompson P. A., Eiseman E., Horak I. D. Expression and interactions of the Src family of tyrosine protein kinases in T lymphocytes. Adv Cancer Res. 1991;57:103–149. doi: 10.1016/s0065-230x(08)60997-5. [DOI] [PubMed] [Google Scholar]
  8. Burkhardt A. L., Brunswick M., Bolen J. B., Mond J. J. Anti-immunoglobulin stimulation of B lymphocytes activates src-related protein-tyrosine kinases. Proc Natl Acad Sci U S A. 1991 Aug 15;88(16):7410–7414. doi: 10.1073/pnas.88.16.7410. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Burkhardt A. L., Stealey B., Rowley R. B., Mahajan S., Prendergast M., Fargnoli J., Bolen J. B. Temporal regulation of non-transmembrane protein tyrosine kinase enzyme activity following T cell antigen receptor engagement. J Biol Chem. 1994 Sep 23;269(38):23642–23647. [PubMed] [Google Scholar]
  10. Cahir McFarland E. D., Hurley T. R., Pingel J. T., Sefton B. M., Shaw A., Thomas M. L. Correlation between Src family member regulation by the protein-tyrosine-phosphatase CD45 and transmembrane signaling through the T-cell receptor. Proc Natl Acad Sci U S A. 1993 Feb 15;90(4):1402–1406. doi: 10.1073/pnas.90.4.1402. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Cambier J. C. New nomenclature for the Reth motif (or ARH1/TAM/ARAM/YXXL) Immunol Today. 1995 Feb;16(2):110–110. doi: 10.1016/0167-5699(95)80105-7. [DOI] [PubMed] [Google Scholar]
  12. Chan A. C., Iwashima M., Turck C. W., Weiss A. ZAP-70: a 70 kd protein-tyrosine kinase that associates with the TCR zeta chain. Cell. 1992 Nov 13;71(4):649–662. doi: 10.1016/0092-8674(92)90598-7. [DOI] [PubMed] [Google Scholar]
  13. Chow L. M., Fournel M., Davidson D., Veillette A. Negative regulation of T-cell receptor signalling by tyrosine protein kinase p50csk. Nature. 1993 Sep 9;365(6442):156–160. doi: 10.1038/365156a0. [DOI] [PubMed] [Google Scholar]
  14. Desai D. M., Sap J., Silvennoinen O., Schlessinger J., Weiss A. The catalytic activity of the CD45 membrane-proximal phosphatase domain is required for TCR signaling and regulation. EMBO J. 1994 Sep 1;13(17):4002–4010. doi: 10.1002/j.1460-2075.1994.tb06716.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Eiseman E., Bolen J. B. Engagement of the high-affinity IgE receptor activates src protein-related tyrosine kinases. Nature. 1992 Jan 2;355(6355):78–80. doi: 10.1038/355078a0. [DOI] [PubMed] [Google Scholar]
  16. Eiseman E., Bolen J. B. Signal transduction by the cytoplasmic domains of Fc epsilon RI-gamma and TCR-zeta in rat basophilic leukemia cells. J Biol Chem. 1992 Oct 15;267(29):21027–21032. [PubMed] [Google Scholar]
  17. Gauen L. K., Zhu Y., Letourneur F., Hu Q., Bolen J. B., Matis L. A., Klausner R. D., Shaw A. S. Interactions of p59fyn and ZAP-70 with T-cell receptor activation motifs: defining the nature of a signalling motif. Mol Cell Biol. 1994 Jun;14(6):3729–3741. doi: 10.1128/mcb.14.6.3729. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Gold M. R., Law D. A., DeFranco A. L. Stimulation of protein tyrosine phosphorylation by the B-lymphocyte antigen receptor. Nature. 1990 Jun 28;345(6278):810–813. doi: 10.1038/345810a0. [DOI] [PubMed] [Google Scholar]
  19. Gounni A. S., Lamkhioued B., Ochiai K., Tanaka Y., Delaporte E., Capron A., Kinet J. P., Capron M. High-affinity IgE receptor on eosinophils is involved in defence against parasites. Nature. 1994 Jan 13;367(6459):183–186. doi: 10.1038/367183a0. [DOI] [PubMed] [Google Scholar]
  20. Gupta S., Weiss A., Kumar G., Wang S., Nel A. The T-cell antigen receptor utilizes Lck, Raf-1, and MEK-1 for activating mitogen-activated protein kinase. Evidence for the existence of a second protein kinase C-dependent pathway in an Lck-negative Jurkat cell mutant. J Biol Chem. 1994 Jun 24;269(25):17349–17357. [PubMed] [Google Scholar]
  21. Hata A., Sabe H., Kurosaki T., Takata M., Hanafusa H. Functional analysis of Csk in signal transduction through the B-cell antigen receptor. Mol Cell Biol. 1994 Nov;14(11):7306–7313. doi: 10.1128/mcb.14.11.7306. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Hirasawa N., Scharenberg A., Yamamura H., Beaven M. A., Kinet J. P. A requirement for Syk in the activation of the microtubule-associated protein kinase/phospholipase A2 pathway by Fc epsilon R1 is not shared by a G protein-coupled receptor. J Biol Chem. 1995 May 5;270(18):10960–10967. doi: 10.1074/jbc.270.18.10960. [DOI] [PubMed] [Google Scholar]
  23. Hovis R. R., Donovan J. A., Musci M. A., Motto D. G., Goldman F. D., Ross S. E., Koretzky G. A. Rescue of signaling by a chimeric protein containing the cytoplasmic domain of CD45. Science. 1993 Apr 23;260(5107):544–546. doi: 10.1126/science.8475387. [DOI] [PubMed] [Google Scholar]
  24. Hurley T. R., Hyman R., Sefton B. M. Differential effects of expression of the CD45 tyrosine protein phosphatase on the tyrosine phosphorylation of the lck, fyn, and c-src tyrosine protein kinases. Mol Cell Biol. 1993 Mar;13(3):1651–1656. doi: 10.1128/mcb.13.3.1651. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Iwashima M., Irving B. A., van Oers N. S., Chan A. C., Weiss A. Sequential interactions of the TCR with two distinct cytoplasmic tyrosine kinases. Science. 1994 Feb 25;263(5150):1136–1139. doi: 10.1126/science.7509083. [DOI] [PubMed] [Google Scholar]
  26. Jouvin M. H., Adamczewski M., Numerof R., Letourneur O., Vallé A., Kinet J. P. Differential control of the tyrosine kinases Lyn and Syk by the two signaling chains of the high affinity immunoglobulin E receptor. J Biol Chem. 1994 Feb 25;269(8):5918–5925. [PubMed] [Google Scholar]
  27. June C. H., Fletcher M. C., Ledbetter J. A., Schieven G. L., Siegel J. N., Phillips A. F., Samelson L. E. Inhibition of tyrosine phosphorylation prevents T-cell receptor-mediated signal transduction. Proc Natl Acad Sci U S A. 1990 Oct;87(19):7722–7726. doi: 10.1073/pnas.87.19.7722. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Kihara H., Siraganian R. P. Src homology 2 domains of Syk and Lyn bind to tyrosine-phosphorylated subunits of the high affinity IgE receptor. J Biol Chem. 1994 Sep 2;269(35):22427–22432. [PubMed] [Google Scholar]
  29. Kinet J. P., Blank U., Ra C., White K., Metzger H., Kochan J. Isolation and characterization of cDNAs coding for the beta subunit of the high-affinity receptor for immunoglobulin E. Proc Natl Acad Sci U S A. 1988 Sep;85(17):6483–6487. doi: 10.1073/pnas.85.17.6483. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Kinet J. P., Metzger H., Hakimi J., Kochan J. A cDNA presumptively coding for the alpha subunit of the receptor with high affinity for immunoglobulin E. Biochemistry. 1987 Jul 28;26(15):4605–4610. doi: 10.1021/bi00389a002. [DOI] [PubMed] [Google Scholar]
  31. Kinet J. P. The gamma-zeta dimers of Fc receptors as connectors to signal transduction. Curr Opin Immunol. 1992 Feb;4(1):43–48. doi: 10.1016/0952-7915(92)90122-u. [DOI] [PubMed] [Google Scholar]
  32. Klausner R. D., Samelson L. E. T cell antigen receptor activation pathways: the tyrosine kinase connection. Cell. 1991 Mar 8;64(5):875–878. doi: 10.1016/0092-8674(91)90310-u. [DOI] [PubMed] [Google Scholar]
  33. Kolanus W., Romeo C., Seed B. T cell activation by clustered tyrosine kinases. Cell. 1993 Jul 16;74(1):171–183. doi: 10.1016/0092-8674(93)90304-9. [DOI] [PubMed] [Google Scholar]
  34. Koretzky G. A., Picus J., Schultz T., Weiss A. Tyrosine phosphatase CD45 is required for T-cell antigen receptor and CD2-mediated activation of a protein tyrosine kinase and interleukin 2 production. Proc Natl Acad Sci U S A. 1991 Mar 15;88(6):2037–2041. doi: 10.1073/pnas.88.6.2037. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Kurosaki T., Takata M., Yamanashi Y., Inazu T., Taniguchi T., Yamamoto T., Yamamura H. Syk activation by the Src-family tyrosine kinase in the B cell receptor signaling. J Exp Med. 1994 May 1;179(5):1725–1729. doi: 10.1084/jem.179.5.1725. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Letourneur F., Klausner R. D. Activation of T cells by a tyrosine kinase activation domain in the cytoplasmic tail of CD3 epsilon. Science. 1992 Jan 3;255(5040):79–82. doi: 10.1126/science.1532456. [DOI] [PubMed] [Google Scholar]
  37. Lin A. Y., Devaux B., Green A., Sagerström C., Elliott J. F., Davis M. M. Expression of T cell antigen receptor heterodimers in a lipid-linked form. Science. 1990 Aug 10;249(4969):677–679. doi: 10.1126/science.1696397. [DOI] [PubMed] [Google Scholar]
  38. Liu F. T., Bohn J. W., Ferry E. L., Yamamoto H., Molinaro C. A., Sherman L. A., Klinman N. R., Katz D. H. Monoclonal dinitrophenyl-specific murine IgE antibody: preparation, isolation, and characterization. J Immunol. 1980 Jun;124(6):2728–2737. [PubMed] [Google Scholar]
  39. Liu X., Brodeur S. R., Gish G., Songyang Z., Cantley L. C., Laudano A. P., Pawson T. Regulation of c-Src tyrosine kinase activity by the Src SH2 domain. Oncogene. 1993 May;8(5):1119–1126. [PubMed] [Google Scholar]
  40. Maurer D., Fiebiger E., Reininger B., Wolff-Winiski B., Jouvin M. H., Kilgus O., Kinet J. P., Stingl G. Expression of functional high affinity immunoglobulin E receptors (Fc epsilon RI) on monocytes of atopic individuals. J Exp Med. 1994 Feb 1;179(2):745–750. doi: 10.1084/jem.179.2.745. [DOI] [PMC free article] [PubMed] [Google Scholar]
  41. Metzger H. The receptor with high affinity for IgE. Immunol Rev. 1992 Feb;125:37–48. doi: 10.1111/j.1600-065x.1992.tb00624.x. [DOI] [PubMed] [Google Scholar]
  42. Motto D. G., Musci M. A., Koretzky G. A. Surface expression of a heterologous phosphatase complements CD45 deficiency in a T cell clone. J Exp Med. 1994 Oct 1;180(4):1359–1366. doi: 10.1084/jem.180.4.1359. [DOI] [PMC free article] [PubMed] [Google Scholar]
  43. Paolini R., Jouvin M. H., Kinet J. P. Phosphorylation and dephosphorylation of the high-affinity receptor for immunoglobulin E immediately after receptor engagement and disengagement. Nature. 1991 Oct 31;353(6347):855–858. doi: 10.1038/353855a0. [DOI] [PubMed] [Google Scholar]
  44. Paolini R., Renard V., Vivier E., Ochiai K., Jouvin M. H., Malissen B., Kinet J. P. Different roles for the Fc epsilon RI gamma chain as a function of the receptor context. J Exp Med. 1995 Jan 1;181(1):247–255. doi: 10.1084/jem.181.1.247. [DOI] [PMC free article] [PubMed] [Google Scholar]
  45. Ravetch J. V. Fc receptors: rubor redux. Cell. 1994 Aug 26;78(4):553–560. doi: 10.1016/0092-8674(94)90521-5. [DOI] [PubMed] [Google Scholar]
  46. Ravetch J. V., Kinet J. P. Fc receptors. Annu Rev Immunol. 1991;9:457–492. doi: 10.1146/annurev.iy.09.040191.002325. [DOI] [PubMed] [Google Scholar]
  47. Reth M. Antigen receptor tail clue. Nature. 1989 Mar 30;338(6214):383–384. doi: 10.1038/338383b0. [DOI] [PubMed] [Google Scholar]
  48. Romeo C., Seed B. Cellular immunity to HIV activated by CD4 fused to T cell or Fc receptor polypeptides. Cell. 1991 Mar 8;64(5):1037–1046. doi: 10.1016/0092-8674(91)90327-u. [DOI] [PubMed] [Google Scholar]
  49. Samelson L. E., Phillips A. F., Luong E. T., Klausner R. D. Association of the fyn protein-tyrosine kinase with the T-cell antigen receptor. Proc Natl Acad Sci U S A. 1990 Jun;87(11):4358–4362. doi: 10.1073/pnas.87.11.4358. [DOI] [PMC free article] [PubMed] [Google Scholar]
  50. Saouaf S. J., Mahajan S., Rowley R. B., Kut S. A., Fargnoli J., Burkhardt A. L., Tsukada S., Witte O. N., Bolen J. B. Temporal differences in the activation of three classes of non-transmembrane protein tyrosine kinases following B-cell antigen receptor surface engagement. Proc Natl Acad Sci U S A. 1994 Sep 27;91(20):9524–9528. doi: 10.1073/pnas.91.20.9524. [DOI] [PMC free article] [PubMed] [Google Scholar]
  51. Shiue L., Green J., Green O. M., Karas J. L., Morgenstern J. P., Ram M. K., Taylor M. K., Zoller M. J., Zydowsky L. D., Bolen J. B. Interaction of p72syk with the gamma and beta subunits of the high-affinity receptor for immunoglobulin E, Fc epsilon RI. Mol Cell Biol. 1995 Jan;15(1):272–281. doi: 10.1128/mcb.15.1.272. [DOI] [PMC free article] [PubMed] [Google Scholar]
  52. Takata M., Sabe H., Hata A., Inazu T., Homma Y., Nukada T., Yamamura H., Kurosaki T. Tyrosine kinases Lyn and Syk regulate B cell receptor-coupled Ca2+ mobilization through distinct pathways. EMBO J. 1994 Mar 15;13(6):1341–1349. doi: 10.1002/j.1460-2075.1994.tb06387.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  53. Takebe Y., Seiki M., Fujisawa J., Hoy P., Yokota K., Arai K., Yoshida M., Arai N. SR alpha promoter: an efficient and versatile mammalian cDNA expression system composed of the simian virus 40 early promoter and the R-U5 segment of human T-cell leukemia virus type 1 long terminal repeat. Mol Cell Biol. 1988 Jan;8(1):466–472. doi: 10.1128/mcb.8.1.466. [DOI] [PMC free article] [PubMed] [Google Scholar]
  54. Takeuchi M., Kuramochi S., Fusaki N., Nada S., Kawamura-Tsuzuku J., Matsuda S., Semba K., Toyoshima K., Okada M., Yamamoto T. Functional and physical interaction of protein-tyrosine kinases Fyn and Csk in the T-cell signaling system. J Biol Chem. 1993 Dec 25;268(36):27413–27419. [PubMed] [Google Scholar]
  55. Taniguchi T., Kobayashi T., Kondo J., Takahashi K., Nakamura H., Suzuki J., Nagai K., Yamada T., Nakamura S., Yamamura H. Molecular cloning of a porcine gene syk that encodes a 72-kDa protein-tyrosine kinase showing high susceptibility to proteolysis. J Biol Chem. 1991 Aug 25;266(24):15790–15796. [PubMed] [Google Scholar]
  56. Timson Gauen L. K., Kong A. N., Samelson L. E., Shaw A. S. p59fyn tyrosine kinase associates with multiple T-cell receptor subunits through its unique amino-terminal domain. Mol Cell Biol. 1992 Dec;12(12):5438–5446. doi: 10.1128/mcb.12.12.5438. [DOI] [PMC free article] [PubMed] [Google Scholar]
  57. Turner J. M., Brodsky M. H., Irving B. A., Levin S. D., Perlmutter R. M., Littman D. R. Interaction of the unique N-terminal region of tyrosine kinase p56lck with cytoplasmic domains of CD4 and CD8 is mediated by cysteine motifs. Cell. 1990 Mar 9;60(5):755–765. doi: 10.1016/0092-8674(90)90090-2. [DOI] [PubMed] [Google Scholar]
  58. Veillette A., Bookman M. A., Horak E. M., Bolen J. B. The CD4 and CD8 T cell surface antigens are associated with the internal membrane tyrosine-protein kinase p56lck. Cell. 1988 Oct 21;55(2):301–308. doi: 10.1016/0092-8674(88)90053-0. [DOI] [PubMed] [Google Scholar]
  59. Volarević S., Niklinska B. B., Burns C. M., June C. H., Weissman A. M., Ashwell J. D. Regulation of TCR signaling by CD45 lacking transmembrane and extracellular domains. Science. 1993 Apr 23;260(5107):541–544. doi: 10.1126/science.8475386. [DOI] [PubMed] [Google Scholar]
  60. Wang B., Rieger A., Kilgus O., Ochiai K., Maurer D., Födinger D., Kinet J. P., Stingl G. Epidermal Langerhans cells from normal human skin bind monomeric IgE via Fc epsilon RI. J Exp Med. 1992 May 1;175(5):1353–1365. doi: 10.1084/jem.175.5.1353. [DOI] [PMC free article] [PubMed] [Google Scholar]
  61. Weiss A., Littman D. R. Signal transduction by lymphocyte antigen receptors. Cell. 1994 Jan 28;76(2):263–274. doi: 10.1016/0092-8674(94)90334-4. [DOI] [PubMed] [Google Scholar]
  62. Weiss A. T cell antigen receptor signal transduction: a tale of tails and cytoplasmic protein-tyrosine kinases. Cell. 1993 Apr 23;73(2):209–212. doi: 10.1016/0092-8674(93)90221-b. [DOI] [PubMed] [Google Scholar]
  63. Yamanashi Y., Kakiuchi T., Mizuguchi J., Yamamoto T., Toyoshima K. Association of B cell antigen receptor with protein tyrosine kinase Lyn. Science. 1991 Jan 11;251(4990):192–194. doi: 10.1126/science.1702903. [DOI] [PubMed] [Google Scholar]
  64. Yi T. L., Bolen J. B., Ihle J. N. Hematopoietic cells express two forms of lyn kinase differing by 21 amino acids in the amino terminus. Mol Cell Biol. 1991 May;11(5):2391–2398. doi: 10.1128/mcb.11.5.2391. [DOI] [PMC free article] [PubMed] [Google Scholar]
  65. Zheng X. M., Wang Y., Pallen C. J. Cell transformation and activation of pp60c-src by overexpression of a protein tyrosine phosphatase. Nature. 1992 Sep 24;359(6393):336–339. doi: 10.1038/359336a0. [DOI] [PubMed] [Google Scholar]
  66. den Hertog J., Pals C. E., Peppelenbosch M. P., Tertoolen L. G., de Laat S. W., Kruijer W. Receptor protein tyrosine phosphatase alpha activates pp60c-src and is involved in neuronal differentiation. EMBO J. 1993 Oct;12(10):3789–3798. doi: 10.1002/j.1460-2075.1993.tb06057.x. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from The EMBO Journal are provided here courtesy of Nature Publishing Group

RESOURCES