Skip to main content
PMC home page
The EMBO Journal logoLink to The EMBO Journal
. 1998 Jul 1;17(13):3704–3713. doi: 10.1093/emboj/17.13.3704

Molecular mechanism of polyhomeotic activation by Engrailed.

N Serrano 1, F Maschat 1
PMCID: PMC1170706  PMID: 9649440

Abstract

The Drosophila Engrailed homeoprotein has been shown to activate directly a Polycomb-group gene, polyhomeotic, during embryogenesis. The molecular mechanism involved in this activation has been studied. Two different types of Engrailed-binding fragments have been detected within the polyhomeotic locus. The P1 and D1 fragments contain several 'TTAATTGCAT' motifs, whereas the D2 fragment contains a long 'TAAT' stretch to which multiple copies of Engrailed bind cooperatively. Another homeodomain-containing protein, Extradenticle, establishes protein-protein interactions with Engrailed on the D2 fragment. We have shown by CAT assays that both types of Engrailed-binding sites (P1 or D1 and D2), as well as Extradenticle, are necessary to obtain activation by Engrailed. In vivo, we have also shown that normal polyhomeotic expression depends on extradenticle expression. Moreover, in the absence of Extradenticle, overexpression of Engrailed protein represses polyhomeotic expression.

Full Text

The Full Text of this article is available as a PDF (419.6 KB).

Selected References

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

  1. Bourbon H. M., Martin-Blanco E., Rosen D., Kornberg T. B. Phosphorylation of the Drosophila engrailed protein at a site outside its homeodomain enhances DNA binding. J Biol Chem. 1995 May 12;270(19):11130–11139. doi: 10.1074/jbc.270.19.11130. [DOI] [PubMed] [Google Scholar]
  2. Chan S. K., Jaffe L., Capovilla M., Botas J., Mann R. S. The DNA binding specificity of Ultrabithorax is modulated by cooperative interactions with extradenticle, another homeoprotein. Cell. 1994 Aug 26;78(4):603–615. doi: 10.1016/0092-8674(94)90525-8. [DOI] [PubMed] [Google Scholar]
  3. Deatrick J., Daly M., Randsholt N. B., Brock H. W. The complex genetic locus polyhomeotic in Drosophila melanogaster potentially encodes two homologous zinc-finger proteins. Gene. 1991 Sep 15;105(2):185–195. doi: 10.1016/0378-1119(91)90150-a. [DOI] [PubMed] [Google Scholar]
  4. Desplan C., Theis J., O'Farrell P. H. The Drosophila developmental gene, engrailed, encodes a sequence-specific DNA binding activity. Nature. 1985 Dec 19;318(6047):630–635. doi: 10.1038/318630a0. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Desplan C., Theis J., O'Farrell P. H. The sequence specificity of homeodomain-DNA interaction. Cell. 1988 Sep 23;54(7):1081–1090. doi: 10.1016/0092-8674(88)90123-7. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Dobens L., Rudolph K., Berger E. M. Ecdysterone regulatory elements function as both transcriptional activators and repressors. Mol Cell Biol. 1991 Apr;11(4):1846–1853. doi: 10.1128/mcb.11.4.1846. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Dura J. M., Randsholt N. B., Deatrick J., Erk I., Santamaria P., Freeman J. D., Freeman S. J., Weddell D., Brock H. W. A complex genetic locus, polyhomeotic, is required for segmental specification and epidermal development in D. melanogaster. Cell. 1987 Dec 4;51(5):829–839. doi: 10.1016/0092-8674(87)90106-1. [DOI] [PubMed] [Google Scholar]
  8. Eaton S., Kornberg T. B. Repression of ci-D in posterior compartments of Drosophila by engrailed. Genes Dev. 1990 Jun;4(6):1068–1077. doi: 10.1101/gad.4.6.1068. [DOI] [PubMed] [Google Scholar]
  9. Fauvarque M. O., Zuber V., Dura J. M. Regulation of polyhomeotic transcription may involve local changes in chromatin activity in Drosophila. Mech Dev. 1995 Aug;52(2-3):343–355. doi: 10.1016/0925-4773(95)00412-t. [DOI] [PubMed] [Google Scholar]
  10. Fjose A., McGinnis W. J., Gehring W. J. Isolation of a homoeo box-containing gene from the engrailed region of Drosophila and the spatial distribution of its transcripts. Nature. 1985 Jan 24;313(6000):284–289. doi: 10.1038/313284a0. [DOI] [PubMed] [Google Scholar]
  11. Gay N. J., Poole S. J., Kornberg T. B. The Drosophila engrailed protein is phosphorylated by a serine-specific protein kinase. Nucleic Acids Res. 1988 Jul 25;16(14A):6637–6647. doi: 10.1093/nar/16.14.6637. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. 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]
  13. Han K., Levine M. S., Manley J. L. Synergistic activation and repression of transcription by Drosophila homeobox proteins. Cell. 1989 Feb 24;56(4):573–583. doi: 10.1016/0092-8674(89)90580-1. [DOI] [PubMed] [Google Scholar]
  14. Han K., Manley J. L. Functional domains of the Drosophila Engrailed protein. EMBO J. 1993 Jul;12(7):2723–2733. doi: 10.1002/j.1460-2075.1993.tb05934.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Heemskerk J., DiNardo S., Kostriken R., O'Farrell P. H. Multiple modes of engrailed regulation in the progression towards cell fate determination. Nature. 1991 Aug 1;352(6334):404–410. doi: 10.1038/352404a0. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Ingham P. W. The molecular genetics of embryonic pattern formation in Drosophila. Nature. 1988 Sep 1;335(6185):25–34. doi: 10.1038/335025a0. [DOI] [PubMed] [Google Scholar]
  17. Jaynes J. B., O'Farrell P. H. Activation and repression of transcription by homoeodomain-containing proteins that bind a common site. Nature. 1988 Dec 22;336(6201):744–749. doi: 10.1038/336744a0. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Jaynes J. B., O'Farrell P. H. Active repression of transcription by the engrailed homeodomain protein. EMBO J. 1991 Jun;10(6):1427–1433. doi: 10.1002/j.1460-2075.1991.tb07663.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. John A., Smith S. T., Jaynes J. B. Inserting the Ftz homeodomain into engrailed creates a dominant transcriptional repressor that specifically turns off Ftz target genes in vivo. Development. 1995 Jun;121(6):1801–1813. doi: 10.1242/dev.121.6.1801. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Kissinger C. R., Liu B. S., Martin-Blanco E., Kornberg T. B., Pabo C. O. Crystal structure of an engrailed homeodomain-DNA complex at 2.8 A resolution: a framework for understanding homeodomain-DNA interactions. Cell. 1990 Nov 2;63(3):579–590. doi: 10.1016/0092-8674(90)90453-l. [DOI] [PubMed] [Google Scholar]
  21. Kornberg T. Compartments in the abdomen of Drosophila and the role of the engrailed locus. Dev Biol. 1981 Sep;86(2):363–372. doi: 10.1016/0012-1606(81)90194-9. [DOI] [PubMed] [Google Scholar]
  22. Kornberg T. Engrailed: a gene controlling compartment and segment formation in Drosophila. Proc Natl Acad Sci U S A. 1981 Feb;78(2):1095–1099. doi: 10.1073/pnas.78.2.1095. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Krasnow M. A., Saffman E. E., Kornfeld K., Hogness D. S. Transcriptional activation and repression by Ultrabithorax proteins in cultured Drosophila cells. Cell. 1989 Jun 16;57(6):1031–1043. doi: 10.1016/0092-8674(89)90341-3. [DOI] [PubMed] [Google Scholar]
  24. Lawrence P. A., Morata G. Compartments in the wing of Drosophila: a study of the engrailed gene. Dev Biol. 1976 Jun;50(2):321–337. doi: 10.1016/0012-1606(76)90155-x. [DOI] [PubMed] [Google Scholar]
  25. Mann R. S., Abu-Shaar M. Nuclear import of the homeodomain protein extradenticle in response to Wg and Dpp signalling. Nature. 1996 Oct 17;383(6601):630–633. doi: 10.1038/383630a0. [DOI] [PubMed] [Google Scholar]
  26. Maxam A. M., Gilbert W. Sequencing end-labeled DNA with base-specific chemical cleavages. Methods Enzymol. 1980;65(1):499–560. doi: 10.1016/s0076-6879(80)65059-9. [DOI] [PubMed] [Google Scholar]
  27. Ohkuma Y., Horikoshi M., Roeder R. G., Desplan C. Engrailed, a homeodomain protein, can repress in vitro transcription by competition with the TATA box-binding protein transcription factor IID. Proc Natl Acad Sci U S A. 1990 Mar;87(6):2289–2293. doi: 10.1073/pnas.87.6.2289. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Peifer M., Wieschaus E. Mutations in the Drosophila gene extradenticle affect the way specific homeo domain proteins regulate segmental identity. Genes Dev. 1990 Jul;4(7):1209–1223. doi: 10.1101/gad.4.7.1209. [DOI] [PubMed] [Google Scholar]
  29. Poole S. J., Kauvar L. M., Drees B., Kornberg T. The engrailed locus of Drosophila: structural analysis of an embryonic transcript. Cell. 1985 Jan;40(1):37–43. doi: 10.1016/0092-8674(85)90306-x. [DOI] [PubMed] [Google Scholar]
  30. Rauskolb C., Wieschaus E. Coordinate regulation of downstream genes by extradenticle and the homeotic selector proteins. EMBO J. 1994 Aug 1;13(15):3561–3569. doi: 10.1002/j.1460-2075.1994.tb06663.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Saenz-Robles M. T., Maschat F., Tabata T., Scott M. P., Kornberg T. B. Selection and characterization of sequences with high affinity for the engrailed protein of Drosophila. Mech Dev. 1995 Oct;53(2):185–195. doi: 10.1016/0925-4773(95)00433-2. [DOI] [PubMed] [Google Scholar]
  32. Serrano N., Brock H. W., Demeret C., Dura J. M., Randsholt N. B., Kornberg T. B., Maschat F. polyhomeotic appears to be a target of engrailed regulation in Drosophila. Development. 1995 Jun;121(6):1691–1703. doi: 10.1242/dev.121.6.1691. [DOI] [PubMed] [Google Scholar]
  33. Serrano N., Brock H. W., Maschat F. beta3-tubulin is directly repressed by the engrailed protein in Drosophila. Development. 1997 Jul;124(13):2527–2536. doi: 10.1242/dev.124.13.2527. [DOI] [PubMed] [Google Scholar]
  34. Sullivan W., Fogarty P., Theurkauf W. Mutations affecting the cytoskeletal organization of syncytial Drosophila embryos. Development. 1993 Aug;118(4):1245–1254. doi: 10.1242/dev.118.4.1245. [DOI] [PubMed] [Google Scholar]
  35. Tabata T., Eaton S., Kornberg T. B. The Drosophila hedgehog gene is expressed specifically in posterior compartment cells and is a target of engrailed regulation. Genes Dev. 1992 Dec;6(12B):2635–2645. doi: 10.1101/gad.6.12b.2635. [DOI] [PubMed] [Google Scholar]
  36. Winslow G. M., Hayashi S., Krasnow M., Hogness D. S., Scott M. P. Transcriptional activation by the Antennapedia and fushi tarazu proteins in cultured Drosophila cells. Cell. 1989 Jun 16;57(6):1017–1030. doi: 10.1016/0092-8674(89)90340-1. [DOI] [PubMed] [Google Scholar]
  37. van Dijk M. A., Murre C. extradenticle raises the DNA binding specificity of homeotic selector gene products. Cell. 1994 Aug 26;78(4):617–624. doi: 10.1016/0092-8674(94)90526-6. [DOI] [PubMed] [Google Scholar]

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

RESOURCES