Abstract
Diptericins are antibacterial polypeptides which are strongly induced in the fat body and blood cells of dipteran insects in response to septic injury. The promoter of the single-copy, intronless diptericin gene of Drosophila contains several nucleotide sequences homologous to mammalian cis-regulatory motifs involved in the control of acute phase response genes. Extending our previous studies on the expression of the diptericin gene, we now report a quantitative analysis of the contribution of various putative regulatory elements to the bacterial inducibility of this gene, based on the generation of 60 transgenic fly lines carrying different elements fused to a reporter gene. Our data definitively identify two Kappa B-related motifs in the proximal promoter as the sites conferring inducibility and tissue-specific expression to the diptericin gene. These motifs alone, however, mediate only minimal levels of expression. Additional proximal regulatory elements are necessary to attain some 20% of the full response and we suspect a role for sequences homologous to mammalian IL6 response elements and interferon-gamma responsive sites in this up-regulation. The transgenic experiments also reveal the existence of a distal regulatory element located upstream of -0.6 kb which increases the level of expression by a factor of five.
Full text
PDFImages in this article
Selected References
These references are in PubMed. This may not be the complete list of references from this article.
- Boman H. G. Antibacterial peptides: key components needed in immunity. Cell. 1991 Apr 19;65(2):205–207. doi: 10.1016/0092-8674(91)90154-q. [DOI] [PubMed] [Google Scholar]
- Bourouis M., Richards G. Remote regulatory sequences of the Drosophila glue gene sgs3 as revealed by P-element transformation. Cell. 1985 Feb;40(2):349–357. doi: 10.1016/0092-8674(85)90149-7. [DOI] [PubMed] [Google Scholar]
- Cociancich S., Bulet P., Hetru C., Hoffmann J. A. The inducible antibacterial peptides of insects. Parasitol Today. 1994 Apr;10(4):132–139. doi: 10.1016/0169-4758(94)90260-7. [DOI] [PubMed] [Google Scholar]
- 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]
- 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]
- Georgel P., Meister M., Kappler C., Lemaitre B., Reichhart J. M., Hoffmann J. A. Insect immunity: the diptericin promoter contains multiple functional regulatory sequences homologous to mammalian acute-phase response elements. Biochem Biophys Res Commun. 1993 Dec 15;197(2):508–517. doi: 10.1006/bbrc.1993.2508. [DOI] [PubMed] [Google Scholar]
- 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]
- Grange T., Roux J., Rigaud G., Pictet R. Cell-type specific activity of two glucocorticoid responsive units of rat tyrosine aminotransferase gene is associated with multiple binding sites for C/EBP and a novel liver-specific nuclear factor. Nucleic Acids Res. 1991 Jan 11;19(1):131–139. doi: 10.1093/nar/19.1.131. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Hiromi Y., Kuroiwa A., Gehring W. J. Control elements of the Drosophila segmentation gene fushi tarazu. Cell. 1985 Dec;43(3 Pt 2):603–613. doi: 10.1016/0092-8674(85)90232-6. [DOI] [PubMed] [Google Scholar]
- 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]
- Hoffmann J. A., Hetru C., Reichhart J. M. The humoral antibacterial response of Drosophila. FEBS Lett. 1993 Jun 28;325(1-2):63–66. doi: 10.1016/0014-5793(93)81414-u. [DOI] [PubMed] [Google Scholar]
- 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]
- 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]
- 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]
- Laval M., Pourrain F., Deutsch J., Lepesant J. A. In vivo functional characterization of an ecdysone response enhancer in the proximal upstream region of the Fbp1 gene of D. melanogaster. Mech Dev. 1993 Dec;44(2-3):123–138. doi: 10.1016/0925-4773(93)90062-3. [DOI] [PubMed] [Google Scholar]
- Lernbecher T., Müller U., Wirth T. Distinct NF-kappa B/Rel transcription factors are responsible for tissue-specific and inducible gene activation. Nature. 1993 Oct 21;365(6448):767–770. doi: 10.1038/365767a0. [DOI] [PubMed] [Google Scholar]
- Martin M., Giangrande A., Ruiz C., Richards G. Induction and repression of the Drosophila Sgs-3 glue gene are mediated by distinct sequences in the proximal promoter. EMBO J. 1989 Feb;8(2):561–568. doi: 10.1002/j.1460-2075.1989.tb03410.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Matsusaka T., Fujikawa K., Nishio Y., Mukaida N., Matsushima K., Kishimoto T., Akira S. Transcription factors NF-IL6 and NF-kappa B synergistically activate transcription of the inflammatory cytokines, interleukin 6 and interleukin 8. Proc Natl Acad Sci U S A. 1993 Nov 1;90(21):10193–10197. doi: 10.1073/pnas.90.21.10193. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Ohmori Y., Hamilton T. A. Cooperative interaction between interferon (IFN) stimulus response element and kappa B sequence motifs controls IFN gamma- and lipopolysaccharide-stimulated transcription from the murine IP-10 promoter. J Biol Chem. 1993 Mar 25;268(9):6677–6688. [PubMed] [Google Scholar]
- 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]
- 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]
- 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]
- Stein B., Baldwin A. S., Jr Distinct mechanisms for regulation of the interleukin-8 gene involve synergism and cooperativity between C/EBP and NF-kappa B. Mol Cell Biol. 1993 Nov;13(11):7191–7198. doi: 10.1128/mcb.13.11.7191. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 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]
- 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]
- Williams B. R. Transcriptional regulation of interferon-stimulated genes. Eur J Biochem. 1991 Aug 15;200(1):1–11. doi: 10.1111/j.1432-1033.1991.tb21041.x. [DOI] [PubMed] [Google Scholar]