Skip to main content
Nucleic Acids Research logoLink to Nucleic Acids Research
. 1997 Mar 15;25(6):1233–1239. doi: 10.1093/nar/25.6.1233

Adjacent GATA and kappa B-like motifs regulate the expression of a Drosophila immune gene.

L Kadalayil 1, U M Petersen 1, Y Engström 1
PMCID: PMC146572  PMID: 9092634

Abstract

The GATA motif is a well known positive cis -regulatory element in vertebrates. In this work we report experimental evidence for the direct participation of a GATA motif in the expression of the Drosophila antibacterial peptide gene Cecropin A1 . Previously we have shown that a kappaB-like site is necessary for Cecropin A1 gene expression. Here we present evidence that the Drosophila Rel protein which binds to the kappaB-like site requires an intact GATA site for maximal Dif-mediated transactivation of the Cecropin A1 gene. We show that a Drosophila blood cell line contains factors binding specifically to the GATA motif of the Cecropin A1 gene. The GATA binding activity is likely to include member(s) of the GATA family of transcriptional regulators. We show that the promoters of several inducible insect immune genes possess GATA sites 0-12 base pairs away from kappaB-like sites in functionally important promoter regions. Clusters of GATA and kappaB sites are also observed in the promoters of two important mammalian immune genes, namely IL6 and IL3. The consistent proximity of GATA and kappaB sites appears to be a common theme in the immune gene expression of insects and mammals.

Full Text

The Full Text of this article is available as a PDF (148.2 KB).

Selected References

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

  1. Abel T., Michelson A. M., Maniatis T. A Drosophila GATA family member that binds to Adh regulatory sequences is expressed in the developing fat body. Development. 1993 Nov;119(3):623–633. doi: 10.1242/dev.119.3.623. [DOI] [PubMed] [Google Scholar]
  2. Ando K., Natori S. Molecular cloning, sequencing, and characterization of cDNA for sarcotoxin IIA, an inducible antibacterial protein of Sarcophaga peregrina (flesh fly). Biochemistry. 1988 Mar 8;27(5):1715–1721. doi: 10.1021/bi00405a050. [DOI] [PubMed] [Google Scholar]
  3. Baeuerle P. A. The inducible transcription activator NF-kappa B: regulation by distinct protein subunits. Biochim Biophys Acta. 1991 Apr 16;1072(1):63–80. doi: 10.1016/0304-419x(91)90007-8. [DOI] [PubMed] [Google Scholar]
  4. Boman H. G. Peptide antibiotics and their role in innate immunity. Annu Rev Immunol. 1995;13:61–92. doi: 10.1146/annurev.iy.13.040195.000425. [DOI] [PubMed] [Google Scholar]
  5. Dushay M. S., Asling B., Hultmark D. Origins of immunity: Relish, a compound Rel-like gene in the antibacterial defense of Drosophila. Proc Natl Acad Sci U S A. 1996 Sep 17;93(19):10343–10347. doi: 10.1073/pnas.93.19.10343. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Engström Y., Kadalayil L., Sun S. C., Samakovlis C., Hultmark D., Faye I. kappa B-like motifs regulate the induction of immune genes in Drosophila. J Mol Biol. 1993 Jul 20;232(2):327–333. doi: 10.1006/jmbi.1993.1392. [DOI] [PubMed] [Google Scholar]
  7. Evans T., Felsenfeld G., Reitman M. Control of globin gene transcription. Annu Rev Cell Biol. 1990;6:95–124. doi: 10.1146/annurev.cb.06.110190.000523. [DOI] [PubMed] [Google Scholar]
  8. Fong T. C., Emerson B. M. The erythroid-specific protein cGATA-1 mediates distal enhancer activity through a specialized beta-globin TATA box. Genes Dev. 1992 Apr;6(4):521–532. doi: 10.1101/gad.6.4.521. [DOI] [PubMed] [Google Scholar]
  9. Freedman L. P., Luisi B. F., Korszun Z. R., Basavappa R., Sigler P. B., Yamamoto K. R. The function and structure of the metal coordination sites within the glucocorticoid receptor DNA binding domain. Nature. 1988 Aug 11;334(6182):543–546. doi: 10.1038/334543a0. [DOI] [PubMed] [Google Scholar]
  10. Fu Y. H., Marzluf G. A. nit-2, the major nitrogen regulatory gene of Neurospora crassa, encodes a protein with a putative zinc finger DNA-binding domain. Mol Cell Biol. 1990 Mar;10(3):1056–1065. doi: 10.1128/mcb.10.3.1056. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Gottschalk L. R., Giannola D. M., Emerson S. G. Molecular regulation of the human IL-3 gene: inducible T cell-restricted expression requires intact AP-1 and Elf-1 nuclear protein binding sites. J Exp Med. 1993 Nov 1;178(5):1681–1692. doi: 10.1084/jem.178.5.1681. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Grant P. A., Arulampalam V., Ahrlund-Richter L., Pettersson S. Identification of Ets-like lymphoid specific elements within the immunoglobulin heavy chain 3' enhancer. Nucleic Acids Res. 1992 Sep 11;20(17):4401–4408. doi: 10.1093/nar/20.17.4401. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Hoffmann J. A. Innate immunity of insects. Curr Opin Immunol. 1995 Feb;7(1):4–10. doi: 10.1016/0952-7915(95)80022-0. [DOI] [PubMed] [Google Scholar]
  14. Hultmark D. Immune reactions in Drosophila and other insects: a model for innate immunity. Trends Genet. 1993 May;9(5):178–183. doi: 10.1016/0168-9525(93)90165-e. [DOI] [PubMed] [Google Scholar]
  15. Hultmark D. Insect immunology. Ancient relationships. Nature. 1994 Jan 13;367(6459):116–117. doi: 10.1038/367116a0. [DOI] [PubMed] [Google Scholar]
  16. Ip Y. T., Levine M. Molecular genetics of Drosophila immunity. Curr Opin Genet Dev. 1994 Oct;4(5):672–677. doi: 10.1016/0959-437x(94)90133-n. [DOI] [PubMed] [Google Scholar]
  17. Ip Y. T., Reach M., Engstrom Y., Kadalayil L., Cai H., González-Crespo S., Tatei K., Levine M. Dif, a dorsal-related gene that mediates an immune response in Drosophila. Cell. 1993 Nov 19;75(4):753–763. doi: 10.1016/0092-8674(93)90495-c. [DOI] [PubMed] [Google Scholar]
  18. Kanai A., Natori S. Analysis of a gene cluster for sarcotoxin II, a group of antibacterial proteins of Sarcophaga peregrina. Mol Cell Biol. 1990 Dec;10(12):6114–6122. doi: 10.1128/mcb.10.12.6114. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Kappler C., Meister M., Lagueux M., Gateff E., Hoffmann J. A., Reichhart J. M. Insect immunity. Two 17 bp repeats nesting a kappa B-related sequence confer inducibility to the diptericin gene and bind a polypeptide in bacteria-challenged Drosophila. EMBO J. 1993 Apr;12(4):1561–1568. doi: 10.1002/j.1460-2075.1993.tb05800.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Kobayashi A., Matsui M., Kubo T., Natori S. Purification and characterization of a 59-kilodalton protein that specifically binds to NF-kappa B-binding motifs of the defense protein genes of Sarcophaga peregrina (the flesh fly). Mol Cell Biol. 1993 Jul;13(7):4049–4056. doi: 10.1128/mcb.13.7.4049. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Kudla B., Caddick M. X., Langdon T., Martinez-Rossi N. M., Bennett C. F., Sibley S., Davies R. W., Arst H. N., Jr The regulatory gene areA mediating nitrogen metabolite repression in Aspergillus nidulans. Mutations affecting specificity of gene activation alter a loop residue of a putative zinc finger. EMBO J. 1990 May;9(5):1355–1364. doi: 10.1002/j.1460-2075.1990.tb08250.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Kylsten P., Samakovlis C., Hultmark D. The cecropin locus in Drosophila; a compact gene cluster involved in the response to infection. EMBO J. 1990 Jan;9(1):217–224. doi: 10.1002/j.1460-2075.1990.tb08098.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Lee M. E., Temizer D. H., Clifford J. A., Quertermous T. Cloning of the GATA-binding protein that regulates endothelin-1 gene expression in endothelial cells. J Biol Chem. 1991 Aug 25;266(24):16188–16192. [PubMed] [Google Scholar]
  24. Lin W. H., Huang L. H., Yeh J. Y., Hoheisel J., Lehrach H., Sun Y. H., Tsai S. F. Expression of a Drosophila GATA transcription factor in multiple tissues in the developing embryos. Identification of homozygous lethal mutants with P-element insertion at the promoter region. J Biol Chem. 1995 Oct 20;270(42):25150–25158. doi: 10.1074/jbc.270.42.25150. [DOI] [PubMed] [Google Scholar]
  25. Lin W. H., Huang L. H., Yeh J. Y., Hoheisel J., Lehrach H., Sun Y. H., Tsai S. F. Expression of a Drosophila GATA transcription factor in multiple tissues in the developing embryos. Identification of homozygous lethal mutants with P-element insertion at the promoter region. J Biol Chem. 1995 Oct 20;270(42):25150–25158. doi: 10.1074/jbc.270.42.25150. [DOI] [PubMed] [Google Scholar]
  26. Orkin S. H. GATA-binding transcription factors in hematopoietic cells. Blood. 1992 Aug 1;80(3):575–581. [PubMed] [Google Scholar]
  27. Petersen U. M., Björklund G., Ip Y. T., Engström Y. The dorsal-related immunity factor, Dif, is a sequence-specific trans-activator of Drosophila Cecropin gene expression. EMBO J. 1995 Jul 3;14(13):3146–3158. doi: 10.1002/j.1460-2075.1995.tb07317.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Ramain P., Heitzler P., Haenlin M., Simpson P. pannier, a negative regulator of achaete and scute in Drosophila, encodes a zinc finger protein with homology to the vertebrate transcription factor GATA-1. Development. 1993 Dec;119(4):1277–1291. doi: 10.1242/dev.119.4.1277. [DOI] [PubMed] [Google Scholar]
  29. Rehorn K. P., Thelen H., Michelson A. M., Reuter R. A molecular aspect of hematopoiesis and endoderm development common to vertebrates and Drosophila. Development. 1996 Dec;122(12):4023–4031. doi: 10.1242/dev.122.12.4023. [DOI] [PubMed] [Google Scholar]
  30. Reichhart J. M., Georgel P., Meister M., Lemaitre B., Kappler C., Hoffmann J. A. Expression and nuclear translocation of the rel/NF-kappa B-related morphogen dorsal during the immune response of Drosophila. C R Acad Sci III. 1993 Oct;316(10):1218–1224. [PubMed] [Google Scholar]
  31. Samakovlis C., Asling B., Boman H. G., Gateff E., Hultmark D. In vitro induction of cecropin genes--an immune response in a Drosophila blood cell line. Biochem Biophys Res Commun. 1992 Nov 16;188(3):1169–1175. doi: 10.1016/0006-291x(92)91354-s. [DOI] [PubMed] [Google Scholar]
  32. Samakovlis C., Kimbrell D. A., Kylsten P., Engström A., Hultmark D. The immune response in Drosophila: pattern of cecropin expression and biological activity. EMBO J. 1990 Sep;9(9):2969–2976. doi: 10.1002/j.1460-2075.1990.tb07489.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Samakovlis C., Kylsten P., Kimbrell D. A., Engström A., Hultmark D. The andropin gene and its product, a male-specific antibacterial peptide in Drosophila melanogaster. EMBO J. 1991 Jan;10(1):163–169. doi: 10.1002/j.1460-2075.1991.tb07932.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Simon M. C. Gotta have GATA. Nat Genet. 1995 Sep;11(1):9–11. doi: 10.1038/ng0995-9. [DOI] [PubMed] [Google Scholar]
  35. Sun S. C., Asling B., Faye I. Organization and expression of the immunoresponsive lysozyme gene in the giant silk moth, Hyalophora cecropia. J Biol Chem. 1991 Apr 5;266(10):6644–6649. [PubMed] [Google Scholar]
  36. Sun S. C., Faye I. Cecropia immunoresponsive factor, an insect immunoresponsive factor with DNA-binding properties similar to nuclear-factor kappa B. Eur J Biochem. 1992 Mar 1;204(2):885–892. doi: 10.1111/j.1432-1033.1992.tb16708.x. [DOI] [PubMed] [Google Scholar]
  37. Sun S. C., Lindström I., Lee J. Y., Faye I. Structure and expression of the attacin genes in Hyalophora cecropia. Eur J Biochem. 1991 Feb 26;196(1):247–254. doi: 10.1111/j.1432-1033.1991.tb15811.x. [DOI] [PubMed] [Google Scholar]
  38. Takahashi H., Komano H., Kawaguchi N., Kitamura N., Nakanishi S., Natori S. Cloning and sequencing of cDNA of Sarcophaga peregrina humoral lectin induced on injury of the body wall. J Biol Chem. 1985 Oct 5;260(22):12228–12233. [PubMed] [Google Scholar]
  39. Taniai K., Ishii T., Sugiyama M., Miyanoshita A., Yamakawa M. Nucleotide sequence of 5'-upstream region and expression of a silkworm gene encoding a new member of the attacin family. Biochem Biophys Res Commun. 1996 Mar 27;220(3):594–599. doi: 10.1006/bbrc.1996.0448. [DOI] [PubMed] [Google Scholar]
  40. Tryselius Y., Samakovlis C., Kimbrell D. A., Hultmark D. CecC, a cecropin gene expressed during metamorphosis in Drosophila pupae. Eur J Biochem. 1992 Feb 15;204(1):395–399. doi: 10.1111/j.1432-1033.1992.tb16648.x. [DOI] [PubMed] [Google Scholar]
  41. Winick J., Abel T., Leonard M. W., Michelson A. M., Chardon-Loriaux I., Holmgren R. A., Maniatis T., Engel J. D. A GATA family transcription factor is expressed along the embryonic dorsoventral axis in Drosophila melanogaster. Development. 1993 Dec;119(4):1055–1065. doi: 10.1242/dev.119.4.1055. [DOI] [PubMed] [Google Scholar]
  42. Yamamoto M., Ko L. J., Leonard M. W., Beug H., Orkin S. H., Engel J. D. Activity and tissue-specific expression of the transcription factor NF-E1 multigene family. Genes Dev. 1990 Oct;4(10):1650–1662. doi: 10.1101/gad.4.10.1650. [DOI] [PubMed] [Google Scholar]
  43. Zhou X., Nguyen T., Kimbrell D. A. Identification and characterization of the Cecropin antibacterial protein gene locus in Drosophila virilis. J Mol Evol. 1997 Mar;44(3):272–281. doi: 10.1007/pl00006144. [DOI] [PubMed] [Google Scholar]
  44. Zon L. I., Tsai S. F., Burgess S., Matsudaira P., Bruns G. A., Orkin S. H. The major human erythroid DNA-binding protein (GF-1): primary sequence and localization of the gene to the X chromosome. Proc Natl Acad Sci U S A. 1990 Jan;87(2):668–672. doi: 10.1073/pnas.87.2.668. [DOI] [PMC free article] [PubMed] [Google Scholar]

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

RESOURCES