Skip to main content
NCBI home page
Nucleic Acids Research logoLink to Nucleic Acids Research
. 1991 Mar 11;19(5):1121–1127. doi: 10.1093/nar/19.5.1121

Cloning and sequence analysis of TFE, a helix-loop-helix transcription factor able to recognize the thyroglobulin gene promoter in vitro.

F Javaux 1, A Donda 1, G Vassart 1, D Christophe 1
PMCID: PMC333790  PMID: 1840650

Abstract

A cDNA that encodes a transcription factor able to recognize the thyroglobulin gene promoter in vitro was isolated from a dog thyroid cDNA expression library in lambda gt11. The library was screened with a multimerized 20 bp-oligonucleotide probe corresponding to the -126 to -107 bp region of the bovine thyroglobulin gene promoter. The specificity of DNA sequence recognition was demonstrated by DNA binding experiments realized with beta-galactosidase-fusion protein immobilized on nitrocellulose filters and various unlabelled multimerized competing DNA fragments. The encoded protein, TFE, appears to be the canine counterpart of a recently cloned human transcription factor, ITF-2, that binds to the mu E5 kappa E2 motif found in both immunoglobulin heavy and light chains genes enhancers and belongs to the basic-Helix-Loop-Helix family of transcription factors. When TFE protein was produced in a rabbit reticulocyte lysate, it displayed the same specificity of DNA sequence recognition as the beta-galactosidase fusion protein and immobilization of the translation product on nitrocellulose still appeared to be essential for detecting in vitro DNA binding activity. Functional data failed to assign a role for TFE in the control of thyroglobulin gene transcription in vitro, suggesting that the selection of TFE clone resulted from the fortuitous presence of a high affinity binding site in the probe used for screening the expression library.

Full text

PDF
1121

Images in this article

1123Image
on p.1123
1125Image
on p.1125
1126Image
on p.1126

Selected References

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

  1. Cai M., Davis R. W. Yeast centromere binding protein CBF1, of the helix-loop-helix protein family, is required for chromosome stability and methionine prototrophy. Cell. 1990 May 4;61(3):437–446. doi: 10.1016/0092-8674(90)90525-j. [DOI] [PubMed] [Google Scholar]
  2. Carr C. S., Sharp P. A. A helix-loop-helix protein related to the immunoglobulin E box-binding proteins. Mol Cell Biol. 1990 Aug;10(8):4384–4388. doi: 10.1128/mcb.10.8.4384. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Christophe D., Gérard C., Juvenal G., Bacolla A., Teugels E., Ledent C., Christophe-Hobertus C., Dumont J. E., Vassart G. Identification of a cAMP-responsive region in thyroglobulin gene promoter. Mol Cell Endocrinol. 1989 Jun;64(1):5–18. doi: 10.1016/0303-7207(89)90060-9. [DOI] [PubMed] [Google Scholar]
  4. Civitareale D., Lonigro R., Sinclair A. J., Di Lauro R. A thyroid-specific nuclear protein essential for tissue-specific expression of the thyroglobulin promoter. EMBO J. 1989 Sep;8(9):2537–2542. doi: 10.1002/j.1460-2075.1989.tb08391.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Davis R. L., Cheng P. F., Lassar A. B., Weintraub H. The MyoD DNA binding domain contains a recognition code for muscle-specific gene activation. Cell. 1990 Mar 9;60(5):733–746. doi: 10.1016/0092-8674(90)90088-v. [DOI] [PubMed] [Google Scholar]
  6. Gonzalez G. A., Montminy M. R. Cyclic AMP stimulates somatostatin gene transcription by phosphorylation of CREB at serine 133. Cell. 1989 Nov 17;59(4):675–680. doi: 10.1016/0092-8674(89)90013-5. [DOI] [PubMed] [Google Scholar]
  7. Gérard C. M., Lefort A., Christophe D., Libert F., Van Sande J., Dumont J. E., Vassart G. Control of thyroperoxidase and thyroglobulin transcription by cAMP: evidence for distinct regulatory mechanisms. Mol Endocrinol. 1989 Dec;3(12):2110–2118. doi: 10.1210/mend-3-12-2110. [DOI] [PubMed] [Google Scholar]
  8. Hansen C., Javaux F., Juvenal G., Vassart G., Christophe D. cAMP-dependent binding of a trans-acting factor to the thyroglobulin promoter. Biochem Biophys Res Commun. 1989 Apr 28;160(2):722–731. doi: 10.1016/0006-291x(89)92493-5. [DOI] [PubMed] [Google Scholar]
  9. Henthorn P., Kiledjian M., Kadesch T. Two distinct transcription factors that bind the immunoglobulin enhancer microE5/kappa 2 motif. Science. 1990 Jan 26;247(4941):467–470. doi: 10.1126/science.2105528. [DOI] [PubMed] [Google Scholar]
  10. Higuchi R., Krummel B., Saiki R. K. A general method of in vitro preparation and specific mutagenesis of DNA fragments: study of protein and DNA interactions. Nucleic Acids Res. 1988 Aug 11;16(15):7351–7367. doi: 10.1093/nar/16.15.7351. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Imagawa M., Chiu R., Karin M. Transcription factor AP-2 mediates induction by two different signal-transduction pathways: protein kinase C and cAMP. Cell. 1987 Oct 23;51(2):251–260. doi: 10.1016/0092-8674(87)90152-8. [DOI] [PubMed] [Google Scholar]
  12. Jones N. Transcriptional regulation by dimerization: two sides to an incestuous relationship. Cell. 1990 Apr 6;61(1):9–11. doi: 10.1016/0092-8674(90)90207-u. [DOI] [PubMed] [Google Scholar]
  13. Kozak M. Point mutations define a sequence flanking the AUG initiator codon that modulates translation by eukaryotic ribosomes. Cell. 1986 Jan 31;44(2):283–292. doi: 10.1016/0092-8674(86)90762-2. [DOI] [PubMed] [Google Scholar]
  14. Lefort A., Lecocq R., Libert F., Lamy F., Swillens S., Vassart G., Dumont J. E. Cloning and sequencing of a calcium-binding protein regulated by cyclic AMP in the thyroid. EMBO J. 1989 Jan;8(1):111–116. doi: 10.1002/j.1460-2075.1989.tb03354.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Murre C., McCaw P. S., Baltimore D. A new DNA binding and dimerization motif in immunoglobulin enhancer binding, daughterless, MyoD, and myc proteins. Cell. 1989 Mar 10;56(5):777–783. doi: 10.1016/0092-8674(89)90682-x. [DOI] [PubMed] [Google Scholar]
  16. Murre C., McCaw P. S., Vaessin H., Caudy M., Jan L. Y., Jan Y. N., Cabrera C. V., Buskin J. N., Hauschka S. D., Lassar A. B. Interactions between heterologous helix-loop-helix proteins generate complexes that bind specifically to a common DNA sequence. Cell. 1989 Aug 11;58(3):537–544. doi: 10.1016/0092-8674(89)90434-0. [DOI] [PubMed] [Google Scholar]
  17. Musti A. M., Ursini V. M., Avvedimento E. V., Zimarino V., Di Lauro R. A cell type specific factor recognizes the rat thyroglobulin promoter. Nucleic Acids Res. 1987 Oct 26;15(20):8149–8166. doi: 10.1093/nar/15.20.8149. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Robertson M. Gene regulation. More to muscle than MyoD. Nature. 1990 Mar 29;344(6265):378–379. doi: 10.1038/344378a0. [DOI] [PubMed] [Google Scholar]
  19. Roesler W. J., Vandenbark G. R., Hanson R. W. Cyclic AMP and the induction of eukaryotic gene transcription. J Biol Chem. 1988 Jul 5;263(19):9063–9066. [PubMed] [Google Scholar]
  20. Rogers S., Wells R., Rechsteiner M. Amino acid sequences common to rapidly degraded proteins: the PEST hypothesis. Science. 1986 Oct 17;234(4774):364–368. doi: 10.1126/science.2876518. [DOI] [PubMed] [Google Scholar]
  21. Seed B., Sheen J. Y. A simple phase-extraction assay for chloramphenicol acyltransferase activity. Gene. 1988 Jul 30;67(2):271–277. doi: 10.1016/0378-1119(88)90403-9. [DOI] [PubMed] [Google Scholar]
  22. Singh H., LeBowitz J. H., Baldwin A. S., Jr, Sharp P. A. Molecular cloning of an enhancer binding protein: isolation by screening of an expression library with a recognition site DNA. Cell. 1988 Feb 12;52(3):415–423. doi: 10.1016/s0092-8674(88)80034-5. [DOI] [PubMed] [Google Scholar]
  23. Van Herle A. J., Vassart G., Dumont J. E. Control of thyroglobulin synthesis and secretion (second of two parts). N Engl J Med. 1979 Aug 9;301(6):307–314. doi: 10.1056/NEJM197908093010605. [DOI] [PubMed] [Google Scholar]
  24. Van Heuverswyn B., Leriche A., Van Sande J., Dumont J. E., Vassart G. Transcriptional control of thyroglobulin gene expression by cyclic AMP. FEBS Lett. 1985 Sep 2;188(2):192–196. doi: 10.1016/0014-5793(85)80370-7. [DOI] [PubMed] [Google Scholar]
  25. Van Heuverswyn B., Streydio C., Brocas H., Refetoff S., Dumont J., Vassart G. Thyrotropin controls transcription of the thyroglobulin gene. Proc Natl Acad Sci U S A. 1984 Oct;81(19):5941–5945. doi: 10.1073/pnas.81.19.5941. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Vinson C. R., LaMarco K. L., Johnson P. F., Landschulz W. H., McKnight S. L. In situ detection of sequence-specific DNA binding activity specified by a recombinant bacteriophage. Genes Dev. 1988 Jul;2(7):801–806. doi: 10.1101/gad.2.7.801. [DOI] [PubMed] [Google Scholar]

Articles from Nucleic Acids Research are provided here courtesy of Oxford University Press

RESOURCES