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. 1991 Jun 15;88(12):5202–5206. doi: 10.1073/pnas.88.12.5202

CD3 eta and CD3 zeta are alternatively spliced products of a common genetic locus and are transcriptionally and/or post-transcriptionally regulated during T-cell development.

L K Clayton 1, L D'Adamio 1, F D Howard 1, M Sieh 1, R E Hussey 1, S Koyasu 1, E L Reinherz 1
PMCID: PMC51840  PMID: 1828894

Abstract

The CD3 eta subunit of the T-cell receptor is thought to subserve an important role in signal transduction and possibly T-cell development. Herein we characterize the organization of the mouse CD3 eta gene and show that it is part of one gene locus that also encodes CD3 zeta on chromosome 1. The NH2-terminal sequence of CD3 zeta and CD3 eta, which share the same leader peptide and are identical through amino acid 122 of each mature protein, is encoded by exons 1-7. However, exons 8 and 9 are differentially spliced to give rise to CD3 zeta and CD3 eta: exons 1-8 encode CD3 zeta and exons 1-7 plus 9 encode CD3 eta. RNase protection analysis with RNA from a variety of fetal, neonatal, and adult cell types indicates that expression of both gene products is T-lineage-restricted. Importantly, expression of CD3 zeta and CD3 eta mRNA appears before or on day 16 of fetal gestation. Expression is apparently coordinate since no cell types tested express CD3 zeta or CD3 eta alone. The steady-state level of CD3 zeta mRNA is greater than or equal to 40-60 times that of CD3 eta mRNA. In immature CD4+CD8+CD3low double-positive thymocytes and CD4+CD8-CD3high or CD4-CD8+CD3high single-positive thymocytes, the respective steady-state CD3 zeta and CD3 eta mRNA levels are equivalent, whereas the amount of receptor-associated CD3 zeta and CD3 eta proteins in double-positive thymocytes is approximately 10 times less than in single-positive thymocytes. Nevertheless, the CD3 zeta/CD3 eta protein ratio remains constant in all populations (40-60:1). Furthermore, discordance between mRNA and protein levels for CD3 zeta and CD3 eta is also observed in splenic T cells. Thus, posttranscriptional and/or transcriptional regulatory mechanisms control CD3 zeta and CD3 eta expression during T-cell development.

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Selected References

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

  1. Ashwell J. D., Klusner R. D. Genetic and mutational analysis of the T-cell antigen receptor. Annu Rev Immunol. 1990;8:139–167. doi: 10.1146/annurev.iy.08.040190.001035. [DOI] [PubMed] [Google Scholar]
  2. Baniyash M., Garcia-Morales P., Bonifacino J. S., Samelson L. E., Klausner R. D. Disulfide linkage of the zeta and eta chains of the T cell receptor. Possible identification of two structural classes of receptors. J Biol Chem. 1988 Jul 15;263(20):9874–9878. [PubMed] [Google Scholar]
  3. Baniyash M., Hsu V. W., Seldin M. F., Klausner R. D. The isolation and characterization of the murine T cell antigen receptor zeta chain gene. J Biol Chem. 1989 Aug 5;264(22):13252–13257. [PubMed] [Google Scholar]
  4. Breathnach R., Chambon P. Organization and expression of eucaryotic split genes coding for proteins. Annu Rev Biochem. 1981;50:349–383. doi: 10.1146/annurev.bi.50.070181.002025. [DOI] [PubMed] [Google Scholar]
  5. Bruce J., Symington F. W., McKearn T. J., Sprent J. A monoclonal antibody discriminating between subsets of T and B cells. J Immunol. 1981 Dec;127(6):2496–2501. [PubMed] [Google Scholar]
  6. Chen C., Bonifacino J. S., Yuan L. C., Klausner R. D. Selective degradation of T cell antigen receptor chains retained in a pre-Golgi compartment. J Cell Biol. 1988 Dec;107(6 Pt 1):2149–2161. doi: 10.1083/jcb.107.6.2149. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Clayton L. K., Bauer A., Jin Y. J., D'Adamio L., Koyasu S., Reinherz E. L. Characterization of thymus-derived lymphocytes expressing Ti alpha-beta CD3 gamma delta epsilon zeta-zeta, Ti alpha-beta CD3 gamma delta epsilon eta-eta or Ti alpha-beta CD3 gamma delta epsilon zeta-zeta/zeta-eta antigen receptor isoforms: analysis by gene transfection. J Exp Med. 1990 Oct 1;172(4):1243–1253. doi: 10.1084/jem.172.4.1243. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Clevers H., Alarcon B., Wileman T., Terhorst C. The T cell receptor/CD3 complex: a dynamic protein ensemble. Annu Rev Immunol. 1988;6:629–662. doi: 10.1146/annurev.iy.06.040188.003213. [DOI] [PubMed] [Google Scholar]
  9. Crispe I. N., Shimonkevitz R. P., Husmann L. A., Kimura J., Allison J. P. Expression of T cell antigen receptor beta-chains on subsets of mouse thymocytes. Analysis by three-color flow cytometry. J Immunol. 1987 Dec 1;139(11):3585–3589. [PubMed] [Google Scholar]
  10. Davis M. M., Bjorkman P. J. T-cell antigen receptor genes and T-cell recognition. Nature. 1988 Aug 4;334(6181):395–402. doi: 10.1038/334395a0. [DOI] [PubMed] [Google Scholar]
  11. Feinberg A. P., Vogelstein B. A technique for radiolabeling DNA restriction endonuclease fragments to high specific activity. Anal Biochem. 1983 Jul 1;132(1):6–13. doi: 10.1016/0003-2697(83)90418-9. [DOI] [PubMed] [Google Scholar]
  12. Hupp K., Siwarski D., Mock B. A., Kinet J. P. Gene mapping of the three subunits of the high affinity FcR for IgE to mouse chromosomes 1 and 19. J Immunol. 1989 Dec 1;143(11):3787–3791. [PubMed] [Google Scholar]
  13. Jin Y. J., Clayton L. K., Howard F. D., Koyasu S., Sieh M., Steinbrich R., Tarr G. E., Reinherz E. L. Molecular cloning of the CD3 eta subunit identifies a CD3 zeta-related product in thymus-derived cells. Proc Natl Acad Sci U S A. 1990 May;87(9):3319–3323. doi: 10.1073/pnas.87.9.3319. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Koyasu S., Nakauchi H., Kitamura K., Yonehara S., Okumura K., Tada T., Yahara I. Production of interleukin 3 and gamma-interferon by an antigen-specific mouse suppressor T cell clone. J Immunol. 1985 May;134(5):3130–3136. [PubMed] [Google Scholar]
  15. Küster H., Thompson H., Kinet J. P. Characterization and expression of the gene for the human Fc receptor gamma subunit. Definition of a new gene family. J Biol Chem. 1990 Apr 15;265(11):6448–6452. [PubMed] [Google Scholar]
  16. Leo O., Foo M., Sachs D. H., Samelson L. E., Bluestone J. A. Identification of a monoclonal antibody specific for a murine T3 polypeptide. Proc Natl Acad Sci U S A. 1987 Mar;84(5):1374–1378. doi: 10.1073/pnas.84.5.1374. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Melton D. A., Krieg P. A., Rebagliati M. R., Maniatis T., Zinn K., Green M. R. Efficient in vitro synthesis of biologically active RNA and RNA hybridization probes from plasmids containing a bacteriophage SP6 promoter. Nucleic Acids Res. 1984 Sep 25;12(18):7035–7056. doi: 10.1093/nar/12.18.7035. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Merćep M., Bonifacino J. S., Garcia-Morales P., Samelson L. E., Klausner R. D., Ashwell J. D. T cell CD3-zeta eta heterodimer expression and coupling to phosphoinositide hydrolysis. Science. 1988 Oct 28;242(4878):571–574. doi: 10.1126/science.2845582. [DOI] [PubMed] [Google Scholar]
  19. Merćep M., Weissman A. M., Frank S. J., Klausner R. D., Ashwell J. D. Activation-driven programmed cell death and T cell receptor zeta eta expression. Science. 1989 Dec 1;246(4934):1162–1165. doi: 10.1126/science.2531464. [DOI] [PubMed] [Google Scholar]
  20. Meuer S. C., Acuto O., Hercend T., Schlossman S. F., Reinherz E. L. The human T-cell receptor. Annu Rev Immunol. 1984;2:23–50. doi: 10.1146/annurev.iy.02.040184.000323. [DOI] [PubMed] [Google Scholar]
  21. Moseley W. S., Seldin M. F. Definition of mouse chromosome 1 and 3 gene linkage groups that are conserved on human chromosome 1: evidence that a conserved linkage group spans the centromere of human chromosome 1. Genomics. 1989 Nov;5(4):899–905. doi: 10.1016/0888-7543(89)90132-8. [DOI] [PubMed] [Google Scholar]
  22. Orloff D. G., Frank S. J., Robey F. A., Weissman A. M., Klausner R. D. Biochemical characterization of the eta chain of the T-cell receptor. A unique subunit related to zeta. J Biol Chem. 1989 Sep 5;264(25):14812–14817. [PubMed] [Google Scholar]
  23. Rackwitz H. R., Zehetner G., Frischauf A. M., Lehrach H. Rapid restriction mapping of DNA cloned in lambda phage vectors. Gene. 1984 Oct;30(1-3):195–200. doi: 10.1016/0378-1119(84)90120-3. [DOI] [PubMed] [Google Scholar]
  24. Ramarli D., Fox D. A., Reinherz E. L. Selective inhibition of interleukin 2 gene function following thymocyte antigen/major histocompatibility complex receptor crosslinking: possible thymic selection mechanism. Proc Natl Acad Sci U S A. 1987 Dec;84(23):8598–8602. doi: 10.1073/pnas.84.23.8598. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Rapp U. R., Cleveland J. L., Brightman K., Scott A., Ihle J. N. Abrogation of IL-3 and IL-2 dependence by recombinant murine retroviruses expressing v-myc oncogenes. Nature. 1985 Oct 3;317(6036):434–438. doi: 10.1038/317434a0. [DOI] [PubMed] [Google Scholar]
  26. Sanger F., Nicklen S., Coulson A. R. DNA sequencing with chain-terminating inhibitors. Proc Natl Acad Sci U S A. 1977 Dec;74(12):5463–5467. doi: 10.1073/pnas.74.12.5463. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Siegelman M. H., Cheng I. C., Weissman I. L., Wakeland E. K. The mouse lymph node homing receptor is identical with the lymphocyte cell surface marker Ly-22: role of the EGF domain in endothelial binding. Cell. 1990 May 18;61(4):611–622. doi: 10.1016/0092-8674(90)90473-r. [DOI] [PubMed] [Google Scholar]
  28. Smith C. A., Williams G. T., Kingston R., Jenkinson E. J., Owen J. J. Antibodies to CD3/T-cell receptor complex induce death by apoptosis in immature T cells in thymic cultures. Nature. 1989 Jan 12;337(6203):181–184. doi: 10.1038/337181a0. [DOI] [PubMed] [Google Scholar]
  29. Southern E. M. Detection of specific sequences among DNA fragments separated by gel electrophoresis. J Mol Biol. 1975 Nov 5;98(3):503–517. doi: 10.1016/s0022-2836(75)80083-0. [DOI] [PubMed] [Google Scholar]
  30. Tan K. N., Datlof B. M., Gilmore J. A., Kronman A. C., Lee J. H., Maxam A. M., Rao A. The T cell receptor V alpha 3 gene segment is associated with reactivity to p-azobenzenearsonate. Cell. 1988 Jul 15;54(2):247–261. doi: 10.1016/0092-8674(88)90557-0. [DOI] [PubMed] [Google Scholar]
  31. Weissman A. M., Baniyash M., Hou D., Samelson L. E., Burgess W. H., Klausner R. D. Molecular cloning of the zeta chain of the T cell antigen receptor. Science. 1988 Feb 26;239(4843):1018–1021. doi: 10.1126/science.3278377. [DOI] [PubMed] [Google Scholar]
  32. Winoto A., Urban J. L., Lan N. C., Goverman J., Hood L., Hansburg D. Predominant use of a V alpha gene segment in mouse T-cell receptors for cytochrome c. Nature. 1986 Dec 18;324(6098):679–682. doi: 10.1038/324679a0. [DOI] [PubMed] [Google Scholar]
  33. Young J. D., Liu C. C., Butler G., Cohn Z. A., Galli S. J. Identification, purification, and characterization of a mast cell-associated cytolytic factor related to tumor necrosis factor. Proc Natl Acad Sci U S A. 1987 Dec;84(24):9175–9179. doi: 10.1073/pnas.84.24.9175. [DOI] [PMC free article] [PubMed] [Google Scholar]

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