Skip to main content

Some NLM-NCBI services and products are experiencing heavy traffic, which may affect performance and availability. We apologize for the inconvenience and appreciate your patience. For assistance, please contact our Help Desk at info@ncbi.nlm.nih.gov.

The EMBO Journal logoLink to The EMBO Journal
. 1992 Apr;11(4):1469–1477. doi: 10.1002/j.1460-2075.1992.tb05191.x

Insect immunity: developmental and inducible activity of the Drosophila diptericin promoter.

J M Reichhart 1, M Meister 1, J L Dimarcq 1, D Zachary 1, D Hoffmann 1, C Ruiz 1, G Richards 1, J A Hoffmann 1
PMCID: PMC556595  PMID: 1373375

Abstract

Diptericins are 9 kDa inducible antibacterial peptides initially isolated from immune haemolymph of Phormia (Diptera). Following the isolation of a Drosophila cDNA encoding a diptericin homologue, we have now cloned a genomic fragment containing the Drosophila diptericin gene. To dissect the regulation of this gene, we have transformed flies with a fusion gene in which the reporter beta-galactosidase gene is under the control of 2.2 kb upstream sequences of the diptericin gene. We show that such a fusion gene is inducible by injection of live bacteria or complete Freund's adjuvant and respects the tissue specific expression pattern of the resident diptericin gene. Our analysis reveals at least four distinct phases in the regulation of this gene: young larvae, late third instar larvae, pupae and adults. This complexity may be related to the presence in the upstream sequences of multiple copies of response elements previously characterized in genes encoding acute phase response proteins in mammals (e.g. NK-kappa B, NF-kappa B related, NF-IL6 response elements).

Full text

PDF
1469

Images in this article

Selected References

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

  1. 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]
  2. 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]
  3. Bulet P., Cociancich S., Dimarcq J. L., Lambert J., Reichhart J. M., Hoffmann D., Hetru C., Hoffmann J. A. Insect immunity. Isolation from a coleopteran insect of a novel inducible antibacterial peptide and of new members of the insect defensin family. J Biol Chem. 1991 Dec 25;266(36):24520–24525. [PubMed] [Google Scholar]
  4. Casteels P., Ampe C., Jacobs F., Vaeck M., Tempst P. Apidaecins: antibacterial peptides from honeybees. EMBO J. 1989 Aug;8(8):2387–2391. doi: 10.1002/j.1460-2075.1989.tb08368.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Dickinson L., Russell V., Dunn P. E. A family of bacteria-regulated, cecropin D-like peptides from Manduca sexta. J Biol Chem. 1988 Dec 25;263(36):19424–19429. [PubMed] [Google Scholar]
  6. Dimarcq J. L., Keppi E., Dunbar B., Lambert J., Reichhart J. M., Hoffmann D., Rankine S. M., Fothergill J. E., Hoffmann J. A. Insect immunity. Purification and characterization of a family of novel inducible antibacterial proteins from immunized larvae of the dipteran Phormia terranovae and complete amino-acid sequence of the predominant member, diptericin A. Eur J Biochem. 1988 Jan 15;171(1-2):17–22. doi: 10.1111/j.1432-1033.1988.tb13752.x. [DOI] [PubMed] [Google Scholar]
  7. Dimarcq J. L., Zachary D., Hoffmann J. A., Hoffmann D., Reichhart J. M. Insect immunity: expression of the two major inducible antibacterial peptides, defensin and diptericin, in Phormia terranovae. EMBO J. 1990 Aug;9(8):2507–2515. doi: 10.1002/j.1460-2075.1990.tb07430.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Faye I., Wyatt G. R. The synthesis of antibacterial proteins in isolated fat body from Cecropia silkmoth pupae. Experientia. 1980 Nov 15;36(11):1325–1326. doi: 10.1007/BF01969615. [DOI] [PubMed] [Google Scholar]
  9. Fowlkes D. M., Mullis N. T., Comeau C. M., Crabtree G. R. Potential basis for regulation of the coordinately expressed fibrinogen genes: homology in the 5' flanking regions. Proc Natl Acad Sci U S A. 1984 Apr;81(8):2313–2316. doi: 10.1073/pnas.81.8.2313. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Georgel P., Ramain P., Giangrande A., Dretzen G., Richards G., Bellard M. Sgs-3 chromatin structure and trans-activators: developmental and ecdysone induction of a glue enhancer-binding factor, GEBF-I, in Drosophila larvae. Mol Cell Biol. 1991 Jan;11(1):523–532. doi: 10.1128/mcb.11.1.523. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Giangrande A., Mettling C., Richards G. Sps-3 transcript levels are determined by multiple remote sequence elements. EMBO J. 1987 Oct;6(10):3079–3084. doi: 10.1002/j.1460-2075.1987.tb02615.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Glaser R. L., Wolfner M. F., Lis J. T. Spatial and temporal pattern of hsp26 expression during normal development. EMBO J. 1986 Apr;5(4):747–754. doi: 10.1002/j.1460-2075.1986.tb04277.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Hattori M., Abraham L. J., Northemann W., Fey G. H. Acute-phase reaction induces a specific complex between hepatic nuclear proteins and the interleukin 6 response element of the rat alpha 2-macroglobulin gene. Proc Natl Acad Sci U S A. 1990 Mar;87(6):2364–2368. doi: 10.1073/pnas.87.6.2364. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Hultmark D., Engström A., Andersson K., Steiner H., Bennich H., Boman H. G. Insect immunity. Attacins, a family of antibacterial proteins from Hyalophora cecropia. EMBO J. 1983;2(4):571–576. doi: 10.1002/j.1460-2075.1983.tb01465.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Hultmark D., Klemenz R., Gehring W. J. Translational and transcriptional control elements in the untranslated leader of the heat-shock gene hsp22. Cell. 1986 Feb 14;44(3):429–438. doi: 10.1016/0092-8674(86)90464-2. [DOI] [PubMed] [Google Scholar]
  16. Isshiki H., Akira S., Sugita T., Nishio Y., Hashimoto S., Pawlowski T., Suematsu S., Kishimoto T. Reciprocal expression of NF-IL6 and C/EBP in hepatocytes: possible involvement of NF-IL6 in acute phase protein gene expression. New Biol. 1991 Jan;3(1):63–70. [PubMed] [Google Scholar]
  17. Kushner I. The phenomenon of the acute phase response. Ann N Y Acad Sci. 1982;389:39–48. doi: 10.1111/j.1749-6632.1982.tb22124.x. [DOI] [PubMed] [Google Scholar]
  18. 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]
  19. Lambert J., Keppi E., Dimarcq J. L., Wicker C., Reichhart J. M., Dunbar B., Lepage P., Van Dorsselaer A., Hoffmann J., Fothergill J. Insect immunity: isolation from immune blood of the dipteran Phormia terranovae of two insect antibacterial peptides with sequence homology to rabbit lung macrophage bactericidal peptides. Proc Natl Acad Sci U S A. 1989 Jan;86(1):262–266. doi: 10.1073/pnas.86.1.262. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Locker J., Buzard G. A dictionary of transcription control sequences. DNA Seq. 1990;1(1):3–11. doi: 10.3109/10425179009041342. [DOI] [PubMed] [Google Scholar]
  21. Matsuyama K., Natori S. Molecular cloning of cDNA for sapecin and unique expression of the sapecin gene during the development of Sarcophaga peregrina. J Biol Chem. 1988 Nov 15;263(32):17117–17121. [PubMed] [Google Scholar]
  22. Morrone G., Ciliberto G., Oliviero S., Arcone R., Dente L., Content J., Cortese R. Recombinant interleukin 6 regulates the transcriptional activation of a set of human acute phase genes. J Biol Chem. 1988 Sep 5;263(25):12554–12558. [PubMed] [Google Scholar]
  23. Murtha M. T., Cavener D. R. Ecdysteroid regulation of glucose dehydrogenase and alcohol dehydrogenase gene expression in Drosophila melanogaster. Dev Biol. 1989 Sep;135(1):66–73. doi: 10.1016/0012-1606(89)90158-9. [DOI] [PubMed] [Google Scholar]
  24. Nanbu R., Nakajima Y., Ando K., Natori S. Novel feature of expression of the sarcotoxin IA gene in development of Sarcophaga peregrina. Biochem Biophys Res Commun. 1988 Jan 29;150(2):540–544. doi: 10.1016/0006-291x(88)90427-5. [DOI] [PubMed] [Google Scholar]
  25. Northemann W., Shiels B. R., Braciak T. A., Hanson R. W., Heinrich P. C., Fey G. H. Structure and acute-phase regulation of the rat alpha 2-macroglobulin gene. Biochemistry. 1988 Dec 27;27(26):9194–9203. doi: 10.1021/bi00426a018. [DOI] [PubMed] [Google Scholar]
  26. Okada M., Natori S. Primary structure of sarcotoxin I, an antibacterial protein induced in the hemolymph of Sarcophaga peregrina (flesh fly) larvae. J Biol Chem. 1985 Jun 25;260(12):7174–7177. [PubMed] [Google Scholar]
  27. Oliviero S., Cortese R. The human haptoglobin gene promoter: interleukin-6-responsive elements interact with a DNA-binding protein induced by interleukin-6. EMBO J. 1989 Apr;8(4):1145–1151. doi: 10.1002/j.1460-2075.1989.tb03485.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Raghavan K. V., Crosby M. A., Mathers P. H., Meyerowitz E. M. Sequences sufficient for correct regulation of Sgs-3 lie close to or within the gene. EMBO J. 1986 Dec 1;5(12):3321–3326. doi: 10.1002/j.1460-2075.1986.tb04646.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Richards G., Cassab A., Bourouis M., Jarry B., Dissous C. The normal developmental regulation of a cloned sgs3 'glue' gene chromosomally integrated in Drosophila melanogaster by P element transformation. EMBO J. 1983;2(12):2137–2142. doi: 10.1002/j.1460-2075.1983.tb01714.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Rubin G. M., Spradling A. C. Vectors for P element-mediated gene transfer in Drosophila. Nucleic Acids Res. 1983 Sep 24;11(18):6341–6351. doi: 10.1093/nar/11.18.6341. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. 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]
  32. Simon J. A., Sutton C. A., Lobell R. B., Glaser R. L., Lis J. T. Determinants of heat shock-induced chromosome puffing. Cell. 1985 Apr;40(4):805–817. doi: 10.1016/0092-8674(85)90340-x. [DOI] [PubMed] [Google Scholar]
  33. Steiner H., Hultmark D., Engström A., Bennich H., Boman H. G. Sequence and specificity of two antibacterial proteins involved in insect immunity. Nature. 1981 Jul 16;292(5820):246–248. doi: 10.1038/292246a0. [DOI] [PubMed] [Google Scholar]
  34. 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]
  35. Urban M. B., Schreck R., Baeuerle P. A. NF-kappa B contacts DNA by a heterodimer of the p50 and p65 subunit. EMBO J. 1991 Jul;10(7):1817–1825. doi: 10.1002/j.1460-2075.1991.tb07707.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Wicker C., Reichhart J. M., Hoffmann D., Hultmark D., Samakovlis C., Hoffmann J. A. Insect immunity. Characterization of a Drosophila cDNA encoding a novel member of the diptericin family of immune peptides. J Biol Chem. 1990 Dec 25;265(36):22493–22498. [PubMed] [Google Scholar]

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

RESOURCES