Skip to main content
PMC home page
Nucleic Acids Research logoLink to Nucleic Acids Research
. 1997 Jun 15;25(12):2293–2300. doi: 10.1093/nar/25.12.2293

Activation of enhancer elements by the homeobox gene Cdx2 is cell line specific.

J K Taylor 1, T Levy 1, E R Suh 1, P G Traber 1
PMCID: PMC146749  PMID: 9171078

Abstract

Cdx2 is a caudal-related homeodomain transcription factor that is expressed in complex patterns during mouse development and at high levels in the intestinal epithelium of adult mice. Cdx2 activates transcription of intestinal gene promoters containing specific binding sites. Moreover, Cdx2 has been shown to induce intestinal differentiation in cell lines. In this study, we show that Cdx2 is able to bind to two well defined enhancer elements in the HoxC8 gene. We then demonstrate that Cdx2 is able to activate transcription of heterologous promoters when its DNA binding element is placed in an enhancer context. Furthermore, the ability to activate enhancer elements is cell-line dependent. When the Cdx2 activation domain was linked to the Gal4 DNA binding domain, the chimeric protein was able to activate Gal4 enhancer constructs in an intestinal cell line, but was unable to activate transcription in NIH3T3 cells. These data suggest that there are cell-specific factors that allow the Cdx2 activation domain to function in the activation of enhancer elements. We hypothesize that either a co-activator protein or differential phosphorylation of the activation domain may be the mechanism for intestinal cell line-specific function of Cdx2 and possibly in other tissues in early development.

Full Text

The Full Text of this article is available as a PDF (155.2 KB).

Selected References

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

  1. Beck F., Erler T., Russell A., James R. Expression of Cdx-2 in the mouse embryo and placenta: possible role in patterning of the extra-embryonic membranes. Dev Dyn. 1995 Nov;204(3):219–227. doi: 10.1002/aja.1002040302. [DOI] [PubMed] [Google Scholar]
  2. Bisaha J. G., Simon T. C., Gordon J. I., Breslow J. L. Characterization of an enhancer element in the human apolipoprotein C-III gene that regulates human apolipoprotein A-I gene expression in the intestinal epithelium. J Biol Chem. 1995 Aug 25;270(34):19979–19988. doi: 10.1074/jbc.270.34.19979. [DOI] [PubMed] [Google Scholar]
  3. Deschamps J., Meijlink F. Mammalian homeobox genes in normal development and neoplasia. Crit Rev Oncog. 1992;3(1-2):117–173. [PubMed] [Google Scholar]
  4. Dignam J. D., Martin P. L., Shastry B. S., Roeder R. G. Eukaryotic gene transcription with purified components. Methods Enzymol. 1983;101:582–598. doi: 10.1016/0076-6879(83)01039-3. [DOI] [PubMed] [Google Scholar]
  5. Drummond F., Sowden J., Morrison K., Edwards Y. H. The caudal-type homeobox protein Cdx-2 binds to the colon promoter of the carbonic anhydrase 1 gene. Eur J Biochem. 1996 Mar 1;236(2):670–681. doi: 10.1111/j.1432-1033.1996.t01-1-00670.x. [DOI] [PubMed] [Google Scholar]
  6. Duprey P., Chowdhury K., Dressler G. R., Balling R., Simon D., Guenet J. L., Gruss P. A mouse gene homologous to the Drosophila gene caudal is expressed in epithelial cells from the embryonic intestine. Genes Dev. 1988 Dec;2(12A):1647–1654. doi: 10.1101/gad.2.12a.1647. [DOI] [PubMed] [Google Scholar]
  7. Dynan W. S. Modularity in promoters and enhancers. Cell. 1989 Jul 14;58(1):1–4. doi: 10.1016/0092-8674(89)90393-0. [DOI] [PubMed] [Google Scholar]
  8. Friedl E. M., Matthias P. Transcriptional activation and repression, two properties of the lymphoid-specific transcription factor Oct-2a. Eur J Biochem. 1995 Nov 15;234(1):308–316. doi: 10.1111/j.1432-1033.1995.308_c.x. [DOI] [PubMed] [Google Scholar]
  9. Gamer L. W., Wright C. V. Murine Cdx-4 bears striking similarities to the Drosophila caudal gene in its homeodomain sequence and early expression pattern. Mech Dev. 1993 Sep;43(1):71–81. doi: 10.1016/0925-4773(93)90024-r. [DOI] [PubMed] [Google Scholar]
  10. Gehring W. J., Affolter M., Bürglin T. Homeodomain proteins. Annu Rev Biochem. 1994;63:487–526. doi: 10.1146/annurev.bi.63.070194.002415. [DOI] [PubMed] [Google Scholar]
  11. German M. S., Wang J., Chadwick R. B., Rutter W. J. Synergistic activation of the insulin gene by a LIM-homeo domain protein and a basic helix-loop-helix protein: building a functional insulin minienhancer complex. Genes Dev. 1992 Nov;6(11):2165–2176. doi: 10.1101/gad.6.11.2165. [DOI] [PubMed] [Google Scholar]
  12. Gordon J. I. Intestinal epithelial differentiation: new insights from chimeric and transgenic mice. J Cell Biol. 1989 Apr;108(4):1187–1194. doi: 10.1083/jcb.108.4.1187. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Gordon J. I., Schmidt G. H., Roth K. A. Studies of intestinal stem cells using normal, chimeric, and transgenic mice. FASEB J. 1992 Sep;6(12):3039–3050. doi: 10.1096/fasebj.6.12.1521737. [DOI] [PubMed] [Google Scholar]
  14. Gorman C. M., Moffat L. F., Howard B. H. Recombinant genomes which express chloramphenicol acetyltransferase in mammalian cells. Mol Cell Biol. 1982 Sep;2(9):1044–1051. doi: 10.1128/mcb.2.9.1044. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Graham F. L., van der Eb A. J. A new technique for the assay of infectivity of human adenovirus 5 DNA. Virology. 1973 Apr;52(2):456–467. doi: 10.1016/0042-6822(73)90341-3. [DOI] [PubMed] [Google Scholar]
  16. Gstaiger M., Georgiev O., van Leeuwen H., van der Vliet P., Schaffner W. The B cell coactivator Bob1 shows DNA sequence-dependent complex formation with Oct-1/Oct-2 factors, leading to differential promoter activation. EMBO J. 1996 Jun 3;15(11):2781–2790. [PMC free article] [PubMed] [Google Scholar]
  17. Gstaiger M., Knoepfel L., Georgiev O., Schaffner W., Hovens C. M. A B-cell coactivator of octamer-binding transcription factors. Nature. 1995 Jan 26;373(6512):360–362. doi: 10.1038/373360a0. [DOI] [PubMed] [Google Scholar]
  18. Harding H. P., Lazar M. A. The monomer-binding orphan receptor Rev-Erb represses transcription as a dimer on a novel direct repeat. Mol Cell Biol. 1995 Sep;15(9):4791–4802. doi: 10.1128/mcb.15.9.4791. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Hu Y., Kazenwadel J., James R. Isolation and characterization of the murine homeobox gene Cdx-1. Regulation of expression in intestinal epithelial cells. J Biol Chem. 1993 Dec 25;268(36):27214–27225. [PubMed] [Google Scholar]
  20. James R., Erler T., Kazenwadel J. Structure of the murine homeobox gene cdx-2. Expression in embryonic and adult intestinal epithelium. J Biol Chem. 1994 May 27;269(21):15229–15237. [PubMed] [Google Scholar]
  21. James R., Kazenwadel J. Homeobox gene expression in the intestinal epithelium of adult mice. J Biol Chem. 1991 Feb 15;266(5):3246–3251. [PubMed] [Google Scholar]
  22. Jin T., Drucker D. J. Activation of proglucagon gene transcription through a novel promoter element by the caudal-related homeodomain protein cdx-2/3. Mol Cell Biol. 1996 Jan;16(1):19–28. doi: 10.1128/mcb.16.1.19. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Lambert M., Colnot S., Suh E., L'Horset F., Blin C., Calliot M. E., Raymondjean M., Thomasset M., Traber P. G., Perret C. cis-Acting elements and transcription factors involved in the intestinal specific expression of the rat calbindin-D9K gene: binding of the intestine-specific transcription factor Cdx-2 to the TATA box. Eur J Biochem. 1996 Mar 15;236(3):778–788. doi: 10.1111/j.1432-1033.1996.00778.x. [DOI] [PubMed] [Google Scholar]
  24. Lillie J. W., Green M. R. Transcription activation by the adenovirus E1a protein. Nature. 1989 Mar 2;338(6210):39–44. doi: 10.1038/338039a0. [DOI] [PubMed] [Google Scholar]
  25. Luo Y., Roeder R. G. Cloning, functional characterization, and mechanism of action of the B-cell-specific transcriptional coactivator OCA-B. Mol Cell Biol. 1995 Aug;15(8):4115–4124. doi: 10.1128/mcb.15.8.4115. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Madden S. L., Cook D. M., Rauscher F. J., 3rd A structure-function analysis of transcriptional repression mediated by the WT1, Wilms' tumor suppressor protein. Oncogene. 1993 Jul;8(7):1713–1720. [PubMed] [Google Scholar]
  27. Meyer B. I., Gruss P. Mouse Cdx-1 expression during gastrulation. Development. 1993 Jan;117(1):191–203. doi: 10.1242/dev.117.1.191. [DOI] [PubMed] [Google Scholar]
  28. Sadowski I., Ptashne M. A vector for expressing GAL4(1-147) fusions in mammalian cells. Nucleic Acids Res. 1989 Sep 25;17(18):7539–7539. doi: 10.1093/nar/17.18.7539. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Schubart D. B., Sauter P., Massa S., Friedl E. M., Schwarzenbach H., Matthias P. Gene structure and characterization of the murine homologue of the B cell-specific transcriptional coactivator OBF-1. Nucleic Acids Res. 1996 May 15;24(10):1913–1920. doi: 10.1093/nar/24.10.1913. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Seipel K., Georgiev O., Schaffner W. Different activation domains stimulate transcription from remote ('enhancer') and proximal ('promoter') positions. EMBO J. 1992 Dec;11(13):4961–4968. doi: 10.1002/j.1460-2075.1992.tb05603.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Shashikant C. S., Bieberich C. J., Belting H. G., Wang J. C., Borbély M. A., Ruddle F. H. Regulation of Hoxc-8 during mouse embryonic development: identification and characterization of critical elements involved in early neural tube expression. Development. 1995 Dec;121(12):4339–4347. doi: 10.1242/dev.121.12.4339. [DOI] [PubMed] [Google Scholar]
  32. Shashikant C. S., Ruddle F. H. Combinations of closely situated cis-acting elements determine tissue-specific patterns and anterior extent of early Hoxc8 expression. Proc Natl Acad Sci U S A. 1996 Oct 29;93(22):12364–12369. doi: 10.1073/pnas.93.22.12364. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Strubin M., Newell J. W., Matthias P. OBF-1, a novel B cell-specific coactivator that stimulates immunoglobulin promoter activity through association with octamer-binding proteins. Cell. 1995 Feb 10;80(3):497–506. doi: 10.1016/0092-8674(95)90500-6. [DOI] [PubMed] [Google Scholar]
  34. Subramanian V., Meyer B. I., Gruss P. Disruption of the murine homeobox gene Cdx1 affects axial skeletal identities by altering the mesodermal expression domains of Hox genes. Cell. 1995 Nov 17;83(4):641–653. doi: 10.1016/0092-8674(95)90104-3. [DOI] [PubMed] [Google Scholar]
  35. Suh E., Chen L., Taylor J., Traber P. G. A homeodomain protein related to caudal regulates intestine-specific gene transcription. Mol Cell Biol. 1994 Nov;14(11):7340–7351. doi: 10.1128/mcb.14.11.7340. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Suh E., Traber P. G. An intestine-specific homeobox gene regulates proliferation and differentiation. Mol Cell Biol. 1996 Feb;16(2):619–625. doi: 10.1128/mcb.16.2.619. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. Traber P. G., Silberg D. G. Intestine-specific gene transcription. Annu Rev Physiol. 1996;58:275–297. doi: 10.1146/annurev.ph.58.030196.001423. [DOI] [PubMed] [Google Scholar]
  38. Traber P. G., Wu G. D., Wang W. Novel DNA-binding proteins regulate intestine-specific transcription of the sucrase-isomaltase gene. Mol Cell Biol. 1992 Aug;12(8):3614–3627. doi: 10.1128/mcb.12.8.3614. [DOI] [PMC free article] [PubMed] [Google Scholar]
  39. Wu G. D., Chen L., Forslund K., Traber P. G. Hepatocyte nuclear factor-1 alpha (HNF-1 alpha) and HNF-1 beta regulate transcription via two elements in an intestine-specific promoter. J Biol Chem. 1994 Jun 24;269(25):17080–17085. [PubMed] [Google Scholar]
  40. Wu G. D., Wang W., Traber P. G. Isolation and characterization of the human sucrase-isomaltase gene and demonstration of intestine-specific transcriptional elements. J Biol Chem. 1992 Apr 15;267(11):7863–7870. [PubMed] [Google Scholar]

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

RESOURCES