Skip to main content
PMC home page
The EMBO Journal logoLink to The EMBO Journal
. 1994 May 1;13(9):2167–2176. doi: 10.1002/j.1460-2075.1994.tb06493.x

Ly-6A is required for T cell receptor expression and protein tyrosine kinase fyn activity.

S K Lee 1, B Su 1, S E Maher 1, A L Bothwell 1
PMCID: PMC395070  PMID: 8187770

Abstract

To characterize the function of the Ly-6A antigen in T cell activation, antisense Ly-6 RNA was expressed in a stably transfected antigen-specific T cell clone. Reduced Ly-6A expression results in inhibition of responses to antigen, anti-TCR (anti-T cell receptor) crosslinking and concanavalin A plus recombinant interleukin 1 and causes impairment of in vitro fyn tyrosine kinase activity. More substantial reduction of Ly-6A results in reduction of TCR expression. Analysis of mRNA species indicates that the reduction is specific for the TCR beta chain. These data demonstrate that Ly-6A may regulate TCR expression and may be involved in early events of T cell activation via regulation of fyn tyrosine kinase activity.

Full text

PDF
2167

Images in this article

2169Image
on p.2169
2171Image
on p.2171
2172Image
on p.2172
2172Image
on p.2172
2172Image
on p.2172
2173Image
on p.2173

Selected References

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

  1. Anderson S. J., Miyake S., Loh D. Y. Transcription from a murine T-cell receptor V beta promoter depends on a conserved decamer motif similar to the cyclic AMP response element. Mol Cell Biol. 1989 Nov;9(11):4835–4845. doi: 10.1128/mcb.9.11.4835. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Auchincloss H., Jr, Ozato K., Sachs D. H. Two distinct murine differentiation antigens determined by genes linked to the Ly-6 locus. J Immunol. 1981 Nov;127(5):1839–1843. [PubMed] [Google Scholar]
  3. Brown D. The tyrosine kinase connection: how GPI-anchored proteins activate T cells. Curr Opin Immunol. 1993 Jun;5(3):349–354. doi: 10.1016/0952-7915(93)90052-t. [DOI] [PubMed] [Google Scholar]
  4. Clevers H., Dunlap S., Saito H., Georgopoulos K., Wileman T., Terhorst C. Characterization and expression of the murine CD3-epsilon gene. Proc Natl Acad Sci U S A. 1988 Nov;85(22):8623–8627. doi: 10.1073/pnas.85.22.8623. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Finkel T. H., Cambier J. C., Kubo R. T., Born W. K., Marrack P., Kappler J. W. The thymus has two functionally distinct populations of immature alpha beta + T cells: one population is deleted by ligation of alpha beta TCR. Cell. 1989 Sep 22;58(6):1047–1054. doi: 10.1016/0092-8674(89)90503-5. [DOI] [PubMed] [Google Scholar]
  6. Finkel T. H., Kappler J. W., Marrack P. C. Immature thymocytes are protected from deletion early in ontogeny. Proc Natl Acad Sci U S A. 1992 Apr 15;89(8):3372–3374. doi: 10.1073/pnas.89.8.3372. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Flood P. M., Dougherty J. P., Ron Y. Inhibition of Ly-6A antigen expression prevents T cell activation. J Exp Med. 1990 Jul 1;172(1):115–120. doi: 10.1084/jem.172.1.115. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Groettrup M., Ungewiss K., Azogui O., Palacios R., Owen M. J., Hayday A. C., von Boehmer H. A novel disulfide-linked heterodimer on pre-T cells consists of the T cell receptor beta chain and a 33 kd glycoprotein. Cell. 1993 Oct 22;75(2):283–294. doi: 10.1016/0092-8674(93)80070-u. [DOI] [PubMed] [Google Scholar]
  9. Janeway C. A., Jr, Carding S., Jones B., Murray J., Portoles P., Rasmussen R., Rojo J., Saizawa K., West J., Bottomly K. CD4+ T cells: specificity and function. Immunol Rev. 1988 Jan;101:39–80. doi: 10.1111/j.1600-065x.1988.tb00732.x. [DOI] [PubMed] [Google Scholar]
  10. June C. H., Fletcher M. C., Ledbetter J. A., Samelson L. E. Increases in tyrosine phosphorylation are detectable before phospholipase C activation after T cell receptor stimulation. J Immunol. 1990 Mar 1;144(5):1591–1599. [PubMed] [Google Scholar]
  11. Karasuyama H., Melchers F. Establishment of mouse cell lines which constitutively secrete large quantities of interleukin 2, 3, 4 or 5, using modified cDNA expression vectors. Eur J Immunol. 1988 Jan;18(1):97–104. doi: 10.1002/eji.1830180115. [DOI] [PubMed] [Google Scholar]
  12. Kaye J., Janeway C. A., Jr The Fab fragment of a directly activating monoclonal antibody that precipitates a disulfide-linked heterodimer from a helper T cell clone blocks activation by either allogeneic Ia or antigen and self-Ia. J Exp Med. 1984 May 1;159(5):1397–1412. doi: 10.1084/jem.159.5.1397. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Kimura S., Tada N., Liu-Lam Y., Hämmerling U. Studies of the mouse Ly-6 alloantigen system. II. Complexities of the Ly-6 region. Immunogenetics. 1984;20(1):47–56. doi: 10.1007/BF00373446. [DOI] [PubMed] [Google Scholar]
  14. Lancki D. W., Ma D. I., Havran W. L., Fitch F. W. Cell surface structures involved in T cell activation. Immunol Rev. 1984 Oct;81:65–94. doi: 10.1111/j.1600-065x.1984.tb01105.x. [DOI] [PubMed] [Google Scholar]
  15. LeClair K. P., Bridgett M. M., Dumont F. J., Palfree R. G., Hämmerling U., Bothwell A. L. Kinetic analysis of Ly-6 gene induction in a T lymphoma by interferons and interleukin 1, and demonstration of Ly-6 inducibility in diverse cell types. Eur J Immunol. 1989 Jul;19(7):1233–1239. doi: 10.1002/eji.1830190713. [DOI] [PubMed] [Google Scholar]
  16. LeClair K. P., Palfree R. G., Flood P. M., Hammerling U., Bothwell A. Isolation of a murine Ly-6 cDNA reveals a new multigene family. EMBO J. 1986 Dec 1;5(12):3227–3234. doi: 10.1002/j.1460-2075.1986.tb04633.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Luo K. X., Sefton B. M. Cross-linking of T-cell surface molecules CD4 and CD8 stimulates phosphorylation of the lck tyrosine protein kinase at the autophosphorylation site. Mol Cell Biol. 1990 Oct;10(10):5305–5313. doi: 10.1128/mcb.10.10.5305. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Owen J. J., Owen M. J., Williams G. T., Kingston R., Jenkinson E. J. The effects of anti-CD3 antibodies on the development of T-cell receptor alpha beta + lymphocytes in embryonic thymus organ cultures. Immunology. 1988 Apr;63(4):639–642. [PMC free article] [PubMed] [Google Scholar]
  19. Palfree R. G., LeClair K. P., Bothwell A., Hämmerling U. cDNA characterization of an Ly-6.2 gene expressed in BW5147 tumor cells. Immunogenetics. 1987;26(6):389–391. doi: 10.1007/BF00343712. [DOI] [PubMed] [Google Scholar]
  20. Palmer D. B., Hayday A., Owen M. J. Is TCR beta expression an essential event in early thymocyte development? Immunol Today. 1993 Sep;14(9):460–462. doi: 10.1016/0167-5699(93)90249-k. [DOI] [PubMed] [Google Scholar]
  21. Petrie H. T., Livak F., Schatz D. G., Strasser A., Crispe I. N., Shortman K. Multiple rearrangements in T cell receptor alpha chain genes maximize the production of useful thymocytes. J Exp Med. 1993 Aug 1;178(2):615–622. doi: 10.1084/jem.178.2.615. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Rock K. L., Reiser H., Bamezai A., McGrew J., Benacerraf B. The LY-6 locus: a multigene family encoding phosphatidylinositol-anchored membrane proteins concerned with T-cell activation. Immunol Rev. 1989 Oct;111:195–224. doi: 10.1111/j.1600-065x.1989.tb00547.x. [DOI] [PubMed] [Google Scholar]
  23. Samelson L. E., Patel M. D., Weissman A. M., Harford J. B., Klausner R. D. Antigen activation of murine T cells induces tyrosine phosphorylation of a polypeptide associated with the T cell antigen receptor. Cell. 1986 Sep 26;46(7):1083–1090. doi: 10.1016/0092-8674(86)90708-7. [DOI] [PubMed] [Google Scholar]
  24. 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]
  25. Shaw A. S., Amrein K. E., Hammond C., Stern D. F., Sefton B. M., Rose J. K. The lck tyrosine protein kinase interacts with the cytoplasmic tail of the CD4 glycoprotein through its unique amino-terminal domain. Cell. 1989 Nov 17;59(4):627–636. doi: 10.1016/0092-8674(89)90008-1. [DOI] [PubMed] [Google Scholar]
  26. Slanetz A. E., Bothwell A. L. Heterodimeric, disulfide-linked alpha/beta T cell receptors in solution. Eur J Immunol. 1991 Jan;21(1):179–183. doi: 10.1002/eji.1830210127. [DOI] [PubMed] [Google Scholar]
  27. Spangrude G. J., Aihara Y., Weissman I. L., Klein J. The stem cell antigens Sca-1 and Sca-2 subdivide thymic and peripheral T lymphocytes into unique subsets. J Immunol. 1988 Dec 1;141(11):3697–3707. [PubMed] [Google Scholar]
  28. Spangrude G. J., Heimfeld S., Weissman I. L. Purification and characterization of mouse hematopoietic stem cells. Science. 1988 Jul 1;241(4861):58–62. doi: 10.1126/science.2898810. [DOI] [PubMed] [Google Scholar]
  29. Stefanová I., Horejsí V., Ansotegui I. J., Knapp W., Stockinger H. GPI-anchored cell-surface molecules complexed to protein tyrosine kinases. Science. 1991 Nov 15;254(5034):1016–1019. doi: 10.1126/science.1719635. [DOI] [PubMed] [Google Scholar]
  30. Straus D. B., Weiss A. Genetic evidence for the involvement of the lck tyrosine kinase in signal transduction through the T cell antigen receptor. Cell. 1992 Aug 21;70(4):585–593. doi: 10.1016/0092-8674(92)90428-f. [DOI] [PubMed] [Google Scholar]
  31. Su B., Waneck G. L., Flavell R. A., Bothwell A. L. The glycosyl phosphatidylinositol anchor is critical for Ly-6A/E-mediated T cell activation. J Cell Biol. 1991 Feb;112(3):377–384. doi: 10.1083/jcb.112.3.377. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Sussman J. J., Merćep M., Saito T., Germain R. N., Bonvini E., Ashwell J. D. Dissociation of phosphoinositide hydrolysis and Ca2+ fluxes from the biological responses of a T-cell hybridoma. Nature. 1988 Aug 18;334(6183):625–628. doi: 10.1038/334625a0. [DOI] [PubMed] [Google Scholar]
  33. Sussman J. J., Saito T., Shevach E. M., Germain R. N., Ashwell J. D. Thy-1- and Ly-6-mediated lymphokine production and growth inhibition of a T cell hybridoma require co-expression of the T cell antigen receptor complex. J Immunol. 1988 Apr 15;140(8):2520–2526. [PubMed] [Google Scholar]
  34. 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]
  35. Thomas P. M., Samelson L. E. The glycophosphatidylinositol-anchored Thy-1 molecule interacts with the p60fyn protein tyrosine kinase in T cells. J Biol Chem. 1992 Jun 15;267(17):12317–12322. [PubMed] [Google Scholar]
  36. 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]
  37. Tsygankov A. Y., Bröker B. M., Fargnoli J., Ledbetter J. A., Bolen J. B. Activation of tyrosine kinase p60fyn following T cell antigen receptor cross-linking. J Biol Chem. 1992 Sep 15;267(26):18259–18262. [PubMed] [Google Scholar]
  38. 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]
  39. Vara J. A., Portela A., Ortín J., Jiménez A. Expression in mammalian cells of a gene from Streptomyces alboniger conferring puromycin resistance. Nucleic Acids Res. 1986 Jun 11;14(11):4617–4624. doi: 10.1093/nar/14.11.4617. [DOI] [PMC free article] [PubMed] [Google Scholar]
  40. Veillette A., Bolen J. B., Bookman M. A. Alterations in tyrosine protein phosphorylation induced by antibody-mediated cross-linking of the CD4 receptor of T lymphocytes. Mol Cell Biol. 1989 Oct;9(10):4441–4446. doi: 10.1128/mcb.9.10.4441. [DOI] [PMC free article] [PubMed] [Google Scholar]
  41. Veillette A., Bookman M. A., Horak E. M., Samelson L. E., Bolen J. B. Signal transduction through the CD4 receptor involves the activation of the internal membrane tyrosine-protein kinase p56lck. Nature. 1989 Mar 16;338(6212):257–259. doi: 10.1038/338257a0. [DOI] [PubMed] [Google Scholar]
  42. Veillette A., Horak I. D., Bolen J. B. Post-translational alterations of the tyrosine kinase p56lck in response to activators of protein kinase C. Oncogene Res. 1988 May;2(4):385–401. [PubMed] [Google Scholar]
  43. Wegener A. M., Letourneur F., Hoeveler A., Brocker T., Luton F., Malissen B. The T cell receptor/CD3 complex is composed of at least two autonomous transduction modules. Cell. 1992 Jan 10;68(1):83–95. doi: 10.1016/0092-8674(92)90208-t. [DOI] [PubMed] [Google Scholar]
  44. Yeh E. T., Reiser H., Bamezai A., Rock K. L. TAP transcription and phosphatidylinositol linkage mutants are defective in activation through the T cell receptor. Cell. 1988 Mar 11;52(5):665–674. doi: 10.1016/0092-8674(88)90404-7. [DOI] [PubMed] [Google Scholar]
  45. Zúiga-Pflücker J. C., Schwartz H. L., Lenardo M. J. Gene transcription in differentiating immature T cell receptor(neg) thymocytes resembles antigen-activated mature T cells. J Exp Med. 1993 Oct 1;178(4):1139–1149. doi: 10.1084/jem.178.4.1139. [DOI] [PMC free article] [PubMed] [Google Scholar]
  46. van de Rijn M., Heimfeld S., Spangrude G. J., Weissman I. L. Mouse hematopoietic stem-cell antigen Sca-1 is a member of the Ly-6 antigen family. Proc Natl Acad Sci U S A. 1989 Jun;86(12):4634–4638. doi: 10.1073/pnas.86.12.4634. [DOI] [PMC free article] [PubMed] [Google Scholar]

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

RESOURCES