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. 1993 Apr;12(4):1561–1568. doi: 10.1002/j.1460-2075.1993.tb05800.x

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.

C Kappler 1, M Meister 1, M Lagueux 1, E Gateff 1, J A Hoffmann 1, J M Reichhart 1
PMCID: PMC413369  PMID: 8467806

Abstract

The Drosophila diptericin gene codes for a 9 kDa antibacterial peptide and is rapidly and transiently expressed in larvae and adults after bacterial challenge. It is also induced in a tumorous Drosophila blood cell line by the addition of lipopolysaccharide (LPS). The promoter of this gene contains two 17 bp repeats located closely upstream of the TATA-box and harbouring a decameric kappa B-related sequence. This study reports that the replacement of the two 17 bp repeats by random sequences abolishes bacteria inducibility in transgenic fly lines. In transfected tumorous blood cells, the replacement of both or either of the 17 bp motifs reduces dramatically LPS inducibility, whereas multiple copies significantly increase the level of transcriptional activation by LPS challenge. A specific DNA-protein binding activity is evidenced in cytoplasmic and nuclear extracts of induced blood cells and fat body. It is absent in controls. It is proposed that induction of the diptericin gene mediated by the two 17 bp repeats occurs via a mechanism similar to that of mammalian NF-kappa B.

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Selected References

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

  1. 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]
  2. Boman H. G., Faye I., Gudmundsson G. H., Lee J. Y., Lidholm D. A. Cell-free immunity in Cecropia. A model system for antibacterial proteins. Eur J Biochem. 1991 Oct 1;201(1):23–31. doi: 10.1111/j.1432-1033.1991.tb16252.x. [DOI] [PubMed] [Google Scholar]
  3. Boshart M., Klüppel M., Schmidt A., Schütz G., Luckow B. Reporter constructs with low background activity utilizing the cat gene. Gene. 1992 Jan 2;110(1):129–130. doi: 10.1016/0378-1119(92)90456-y. [DOI] [PubMed] [Google Scholar]
  4. Dignam J. D., Lebovitz R. M., Roeder R. G. Accurate transcription initiation by RNA polymerase II in a soluble extract from isolated mammalian nuclei. Nucleic Acids Res. 1983 Mar 11;11(5):1475–1489. doi: 10.1093/nar/11.5.1475. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. 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]
  6. Eisenberg S., Francesconi S. C., Civalier C., Walker S. S. Purification of DNA-binding proteins by site-specific DNA affinity chromatography. Methods Enzymol. 1990;182:521–529. doi: 10.1016/0076-6879(90)82041-y. [DOI] [PubMed] [Google Scholar]
  7. Engels W. R., Johnson-Schlitz D. M., Eggleston W. B., Sved J. High-frequency P element loss in Drosophila is homolog dependent. Cell. 1990 Aug 10;62(3):515–525. doi: 10.1016/0092-8674(90)90016-8. [DOI] [PubMed] [Google Scholar]
  8. Gateff E. Malignant neoplasms of genetic origin in Drosophila melanogaster. Science. 1978 Jun 30;200(4349):1448–1459. doi: 10.1126/science.96525. [DOI] [PubMed] [Google Scholar]
  9. 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]
  10. 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]
  11. Herbomel P., Bourachot B., Yaniv M. Two distinct enhancers with different cell specificities coexist in the regulatory region of polyoma. Cell. 1984 Dec;39(3 Pt 2):653–662. doi: 10.1016/0092-8674(84)90472-0. [DOI] [PubMed] [Google Scholar]
  12. Hocke G. M., Barry D., Fey G. H. Synergistic action of interleukin-6 and glucocorticoids is mediated by the interleukin-6 response element of the rat alpha 2 macroglobulin gene. Mol Cell Biol. 1992 May;12(5):2282–2294. doi: 10.1128/mcb.12.5.2282. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Hoffmann J. A., Hetru C. Insect defensins: inducible antibacterial peptides. Immunol Today. 1992 Oct;13(10):411–415. doi: 10.1016/0167-5699(92)90092-L. [DOI] [PubMed] [Google Scholar]
  14. 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]
  15. Kieny M. P., Lathe R., Lecocq J. P. New versatile cloning and sequencing vectors based on bacteriophage M13. Gene. 1983 Dec;26(1):91–99. doi: 10.1016/0378-1119(83)90039-2. [DOI] [PubMed] [Google Scholar]
  16. 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]
  17. Lenardo M. J., Baltimore D. NF-kappa B: a pleiotropic mediator of inducible and tissue-specific gene control. Cell. 1989 Jul 28;58(2):227–229. doi: 10.1016/0092-8674(89)90833-7. [DOI] [PubMed] [Google Scholar]
  18. Reichhart J. M., Essrich M., Dimarcq J. L., Hoffmann D., Hoffmann J. A., Lagueux M. Insect immunity. Isolation of cDNA clones corresponding to diptericin, an inducible antibacterial peptide from Phormia terranovae (Diptera). Transcriptional profiles during immunization. Eur J Biochem. 1989 Jun 15;182(2):423–427. doi: 10.1111/j.1432-1033.1989.tb14848.x. [DOI] [PubMed] [Google Scholar]
  19. Reichhart J. M., Meister M., Dimarcq J. L., Zachary D., Hoffmann D., Ruiz C., Richards G., Hoffmann J. A. Insect immunity: developmental and inducible activity of the Drosophila diptericin promoter. EMBO J. 1992 Apr;11(4):1469–1477. doi: 10.1002/j.1460-2075.1992.tb05191.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. 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]
  21. Sen R., Baltimore D. Multiple nuclear factors interact with the immunoglobulin enhancer sequences. Cell. 1986 Aug 29;46(5):705–716. doi: 10.1016/0092-8674(86)90346-6. [DOI] [PubMed] [Google Scholar]
  22. Steward R. Dorsal, an embryonic polarity gene in Drosophila, is homologous to the vertebrate proto-oncogene, c-rel. Science. 1987 Oct 30;238(4827):692–694. doi: 10.1126/science.3118464. [DOI] [PubMed] [Google Scholar]
  23. 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]
  24. 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]
  25. 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]
  26. Thisse C., Perrin-Schmitt F., Stoetzel C., Thisse B. Sequence-specific transactivation of the Drosophila twist gene by the dorsal gene product. Cell. 1991 Jun 28;65(7):1191–1201. doi: 10.1016/0092-8674(91)90014-p. [DOI] [PubMed] [Google Scholar]
  27. 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]

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