Skip to main content
Molecular and Cellular Biology logoLink to Molecular and Cellular Biology
. 1993 Mar;13(3):1943–1950. doi: 10.1128/mcb.13.3.1943

Extinction of T cell receptor alpha-chain gene expression accompanied by loss of the lymphoid enhancer-binding factor 1 (LEF-1) in murine somatic cell hybrids.

T Yamada 1, Y Hitomi 1, K Shimizu 1, M Ohki 1, T Oikawa 1
PMCID: PMC359508  PMID: 8382779

Abstract

To investigate the presence of a negative regulatory factor(s) suppressing T-cell receptor alpha-chain (TCR alpha) gene expression in non-T cells, 10 independent cell hybrid clones were generated between mouse T-cell lymphoma EL4 cells (TCR alpha+/beta+) and mouse fibroblast B82 cells. These cell hybrids showed a typical fibroblastic morphology and retained an approximate sum of chromosome numbers derived from both parental cells. No transcripts of the TCR alpha gene were detected in the cell hybrids, although the presence of the rearranged TCR alpha allele from EL4 cells was confirmed. The possibility of involvement of nuclear proteins responsible for the activity of the TCR alpha gene enhancer in the extinction of TCR alpha gene expression in the cell hybrids was examined. Nuclear proteins which bind to the lymphoid enhancer-binding factor 1 (LEF-1) binding motif present in EL4 cells disappeared in the hybrid clones, whereas no significant change was observed in DNA-binding activity of nuclear proteins to a consensus cyclic AMP response element (CRE) and the Ets-1 binding motif between the parental cells and the cell hybrids. No transcripts of the LEF-1 gene were detected in the cell hybrids, despite the retention of the LEF-1 gene and murine chromosomes 3, on which the LEF-1 allele is located, from both parental cells. These results suggest that a trans-acting negative regulatory factor(s) present in fibroblasts suppresses LEF-1 gene expression and that suppression of LEF-1 may lead to the extinction of TCR alpha gene expression in the cell hybrids.

Full text

PDF
1943

Images in this article

Selected References

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

  1. Bergman Y., Strich B., Sharir H., Ber R., Laskov R. Extinction of Ig genes expression in myeloma x fibroblast somatic cell hybrids is accompanied by repression of the oct-2 gene encoding a B-cell specific transcription factor. EMBO J. 1990 Mar;9(3):849–855. doi: 10.1002/j.1460-2075.1990.tb08182.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Besnard C., Lores P., Daegelen D., Jami J. Reversible extinction of insulin gene expression in insulinoma x fibroblast somatic cell hybrids. Exp Cell Res. 1989 Nov;185(1):101–108. doi: 10.1016/0014-4827(89)90040-2. [DOI] [PubMed] [Google Scholar]
  3. Bhat N. K., Fisher R. J., Fujiwara S., Ascione R., Papas T. S. Temporal and tissue-specific expression of mouse ets genes. Proc Natl Acad Sci U S A. 1987 May;84(10):3161–3165. doi: 10.1073/pnas.84.10.3161. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Bhat N. K., Komschlies K. L., Fujiwara S., Fisher R. J., Mathieson B. J., Gregorio T. A., Young H. A., Kasik J. W., Ozato K., Papas T. S. Expression of ets genes in mouse thymocyte subsets and T cells. J Immunol. 1989 Jan 15;142(2):672–678. [PubMed] [Google Scholar]
  5. Boshart M., Weih F., Nichols M., Schütz G. The tissue-specific extinguisher locus TSE1 encodes a regulatory subunit of cAMP-dependent protein kinase. Cell. 1991 Sep 6;66(5):849–859. doi: 10.1016/0092-8674(91)90432-x. [DOI] [PubMed] [Google Scholar]
  6. Cornall R. J., Aitman T. J., Hearne C. M., Todd J. A. The generation of a library of PCR-analyzed microsatellite variants for genetic mapping of the mouse genome. Genomics. 1991 Aug;10(4):874–881. doi: 10.1016/0888-7543(91)90175-e. [DOI] [PubMed] [Google Scholar]
  7. Dignam J. D., Lebovitz R. M., Roeder R. G. Accurate transcription initiation by RNA polymerase II in a soluble extract from isolated mammalian nuclei. Nucleic Acids Res. 1983 Mar 11;11(5):1475–1489. doi: 10.1093/nar/11.5.1475. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Giese K., Amsterdam A., Grosschedl R. DNA-binding properties of the HMG domain of the lymphoid-specific transcriptional regulator LEF-1. Genes Dev. 1991 Dec;5(12B):2567–2578. doi: 10.1101/gad.5.12b.2567. [DOI] [PubMed] [Google Scholar]
  9. Greenberg A., Ber R., Kra-Oz Z., Laskov R. Extinction of expression of immunoglobulin genes in myeloma X fibroblast somatic cell hybrids. Mol Cell Biol. 1987 Feb;7(2):936–939. doi: 10.1128/mcb.7.2.936. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Gunning P., Ponte P., Kedes L., Hickey R. J., Skoultchi A. I. Expression of human cardiac actin in mouse L cells: a sarcomeric actin associates with a nonmuscle cytoskeleton. Cell. 1984 Mar;36(3):709–715. doi: 10.1016/0092-8674(84)90351-9. [DOI] [PubMed] [Google Scholar]
  11. Ho I. C., Bhat N. K., Gottschalk L. R., Lindsten T., Thompson C. B., Papas T. S., Leiden J. M. Sequence-specific binding of human Ets-1 to the T cell receptor alpha gene enhancer. Science. 1990 Nov 9;250(4982):814–818. doi: 10.1126/science.2237431. [DOI] [PubMed] [Google Scholar]
  12. Ho I. C., Leiden J. M. Regulation of the human T-cell receptor alpha gene enhancer: multiple ubiquitous and T-cell-specific nuclear proteins interact with four hypomethylated enhancer elements. Mol Cell Biol. 1990 Sep;10(9):4720–4727. doi: 10.1128/mcb.10.9.4720. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Ho I. C., Leiden J. M. The T alpha 2 nuclear protein binding site from the human T cell receptor alpha enhancer functions as both a T cell-specific transcriptional activator and repressor. J Exp Med. 1990 Nov 1;172(5):1443–1449. doi: 10.1084/jem.172.5.1443. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Ho I. C., Yang L. H., Morle G., Leiden J. M. A T-cell-specific transcriptional enhancer element 3' of C alpha in the human T-cell receptor alpha locus. Proc Natl Acad Sci U S A. 1989 Sep;86(17):6714–6718. doi: 10.1073/pnas.86.17.6714. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Iwamoto A., Ohashi P. S., Pircher H., Walker C. L., Michalopoulos E. E., Rupp F., Hengartner H., Mak T. W. T cell receptor variable gene usage in a specific cytotoxic T cell response. Primary structure of the antigen-MHC receptor of four hapten-specific cytotoxic T cell clones. J Exp Med. 1987 Mar 1;165(3):591–600. doi: 10.1084/jem.165.3.591. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Jones K. W., Shapero M. H., Chevrette M., Fournier R. E. Subtractive hybridization cloning of a tissue-specific extinguisher: TSE1 encodes a regulatory subunit of protein kinase A. Cell. 1991 Sep 6;66(5):861–872. doi: 10.1016/0092-8674(91)90433-y. [DOI] [PubMed] [Google Scholar]
  17. Junker S., Pedersen S., Schreiber E., Matthias P. Extinction of an immunoglobulin kappa promoter in cell hybrids is mediated by the octamer motif and correlates with suppression of Oct-2 expression. Cell. 1990 May 4;61(3):467–474. doi: 10.1016/0092-8674(90)90528-m. [DOI] [PubMed] [Google Scholar]
  18. Karpinski B. A., Yang L. H., Cacheris P., Morle G. D., Leiden J. M. The first intron of the 4F2 heavy-chain gene contains a transcriptional enhancer element that binds multiple nuclear proteins. Mol Cell Biol. 1989 Jun;9(6):2588–2597. doi: 10.1128/mcb.9.6.2588. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Leshkowitz D., Walker M. D. Extinction of insulin gene expression in hybrids between beta cells and fibroblasts is accompanied by loss of the putative beta-cell-specific transcription factor IEF1. Mol Cell Biol. 1991 Mar;11(3):1547–1552. doi: 10.1128/mcb.11.3.1547. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. MacLeod C. L., Minning L., Gold D. P., Terhorst C., Wilkinson M. Negative trans-regulation of T-cell antigen receptor/T3 complex mRNA expression in murine T-lymphoma somatic cell hybrids. Proc Natl Acad Sci U S A. 1986 Sep;83(18):6989–6993. doi: 10.1073/pnas.83.18.6989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Maeda K., Nakashima M., Komori S., Watanabe T. Trans-acting regulatory factors for T cell antigen receptor alpha- and gamma-chain gene expression. J Immunol. 1988 Apr 15;140(8):2796–2801. [PubMed] [Google Scholar]
  22. Martin M. E., Piette J., Yaniv M., Tang W. J., Folk W. R. Activation of the polyomavirus enhancer by a murine activator protein 1 (AP1) homolog and two contiguous proteins. Proc Natl Acad Sci U S A. 1988 Aug;85(16):5839–5843. doi: 10.1073/pnas.85.16.5839. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. McCormick A., Wu D., Castrillo J. L., Dana S., Strobl J., Thompson E. B., Karin M. Extinction of growth hormone expression in somatic cell hybrids involves repression of the specific trans-activator GHF-1. Cell. 1988 Oct 21;55(2):379–389. doi: 10.1016/0092-8674(88)90061-x. [DOI] [PubMed] [Google Scholar]
  24. Milatovich A., Travis A., Grosschedl R., Francke U. Gene for lymphoid enhancer-binding factor 1 (LEF1) mapped to human chromosome 4 (q23-q25) and mouse chromosome 3 near Egf. Genomics. 1991 Dec;11(4):1040–1048. doi: 10.1016/0888-7543(91)90030-i. [DOI] [PubMed] [Google Scholar]
  25. Müller M. M., Ruppert S., Schaffner W., Matthias P. A cloned octamer transcription factor stimulates transcription from lymphoid-specific promoters in non-B cells. Nature. 1988 Dec 8;336(6199):544–551. doi: 10.1038/336544a0. [DOI] [PubMed] [Google Scholar]
  26. Nishikura K., ar-Rushdi A., Erikson J., DeJesus E., Dugan D., Croce C. M. Repression of rearranged mu gene and translocated c-myc in mouse 3T3 cells X Burkitt lymphoma cell hybrids. Science. 1984 Apr 27;224(4647):399–402. doi: 10.1126/science.6424234. [DOI] [PubMed] [Google Scholar]
  27. Oikawa T., Kondoh N., Fujita H., Satoh C., Li S. Z., Yoshida M. C., Imamura M., Onoé K., Kuzumaki N. Transcriptional down-regulation of the rearranged C-myc expression in murine cell hybrids between a plasmacytoma and a T-cell lymphoma. Int J Cancer. 1990 Mar 15;45(3):468–474. doi: 10.1002/ijc.2910450316. [DOI] [PubMed] [Google Scholar]
  28. Scheidereit C., Cromlish J. A., Gerster T., Kawakami K., Balmaceda C. G., Currie R. A., Roeder R. G. A human lymphoid-specific transcription factor that activates immunoglobulin genes is a homoeobox protein. Nature. 1988 Dec 8;336(6199):551–557. doi: 10.1038/336551a0. [DOI] [PubMed] [Google Scholar]
  29. Staudt L. M., Clerc R. G., Singh H., LeBowitz J. H., Sharp P. A., Baltimore D. Cloning of a lymphoid-specific cDNA encoding a protein binding the regulatory octamer DNA motif. Science. 1988 Jul 29;241(4865):577–580. doi: 10.1126/science.3399892. [DOI] [PubMed] [Google Scholar]
  30. Waterman M. L., Fischer W. H., Jones K. A. A thymus-specific member of the HMG protein family regulates the human T cell receptor C alpha enhancer. Genes Dev. 1991 Apr;5(4):656–669. doi: 10.1101/gad.5.4.656. [DOI] [PubMed] [Google Scholar]
  31. Waterman M. L., Jones K. A. Purification of TCF-1 alpha, a T-cell-specific transcription factor that activates the T-cell receptor C alpha gene enhancer in a context-dependent manner. New Biol. 1990 Jul;2(7):621–636. [PubMed] [Google Scholar]
  32. Winoto A., Baltimore D. A novel, inducible and T cell-specific enhancer located at the 3' end of the T cell receptor alpha locus. EMBO J. 1989 Mar;8(3):729–733. doi: 10.1002/j.1460-2075.1989.tb03432.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Winoto A., Baltimore D. Alpha beta lineage-specific expression of the alpha T cell receptor gene by nearby silencers. Cell. 1989 Nov 17;59(4):649–655. doi: 10.1016/0092-8674(89)90010-x. [DOI] [PubMed] [Google Scholar]
  34. Yamada T., Sasaki M., Yoshida M. C., Aya T., Koizumi S., Osato T. Establishment and characterization of Ph1-positive and Ph1-negative lymphoblastoid cell lines from a patient with chronic myelogenous leukemia. Jpn J Cancer Res. 1985 May;76(5):365–373. [PubMed] [Google Scholar]
  35. Yu H., Porton B., Shen L. Y., Eckhardt L. A. Role of the octamer motif in hybrid cell extinction of immunoglobulin gene expression: extinction is dominant in a two enhancer system. Cell. 1989 Aug 11;58(3):441–448. doi: 10.1016/0092-8674(89)90425-x. [DOI] [PubMed] [Google Scholar]

Articles from Molecular and Cellular Biology are provided here courtesy of Taylor & Francis

RESOURCES