A Large-Scale RNAi Screen Identifies Deaf1 as a Regulator of Innate Immune Responses in DrosophilaKuttenkeuler D.a · Pelte N.a · Ragab A.a · Gesellchen V.a · Schneider L.a · Blass C.a · Axelsson E.b · Huber W.b · Boutros M.a
aGerman Cancer Research Center (DKFZ) and University of Heidelberg, Division of Signaling and Functional Genomics, University of Heidelberg, Heidelberg, Germany; bEuropean Molecular Biology Laboratory, European Bioinformatics Institute, Cambridge, UK
Prof. Dr. Michael Boutros
German Cancer Research Center (DKFZ) and University of Heidelberg
Im Neuenheimer Feld 580
DE–69120 Heidelberg (Germany)
Tel. +49 6221 42 1951, Fax +49 6221 42 1959, E-Mail email@example.com
Do you have an account?
Innate immune signalling pathways are evolutionarily conserved between invertebrates and vertebrates. The analysis of NF-ĸB signalling in Drosophila has contributed important insights into how organisms respond to infection. Nevertheless, significant gaps remain in our understanding of how the activation of intracellular signalling elicits specific transcriptional programs. Here we report a genome-wide RNA interference survey for transcription factors that are required for Toll-dependent immune responses. In addition to the NF-ĸB homologs Dif, Dorsal and factors of the general transcription machinery, we identified Deformed Epidermal Autoregulatory Factor 1 (Deaf1) to be required for the expression of the Toll target gene Drosomycin in cultured cells and in Drosophila in vivo. We show that Deaf1 is required for the survival of flies after fungal, but not E. coli, infection. We determine that Deaf1 acts downstream of the NF-ĸB factors Dorsal and Dif. These results indicate that Deaf1 is an important contributor to innate immune responses in vivo.
© 2009 S. Karger AG, Basel
- Lemaitre B, Hoffmann J: The host defense of Drosophila melanogaster. Annu Rev Immunol 2007;25:697–743.
- Ferrandon D, Imler JL, Hetru C, Hoffmann JA: The Drosophila systemic immuneresponse: sensing and signalling during bacterial and fungal infections. Nat Rev Immunol 2007;7:862–874.
- Jang IH, Chosa N, Kim SH, Nam HJ, Lemaitre B, Ochiai M, Kambris Z, Brun S, Hashimoto C, Ashida M, Brey PT, Lee WJ: A Spätzle-processing enzyme required for Toll signaling activation in Drosophila innate immunity. Dev Cell 2006;10:45–55.
- El Chamy L, Leclerc V, Caldelari I, Reichhart JM: Sensing of ‘danger signals’ and pathogen-associated molecular patterns defines binary signaling pathways ‘upstream’ of Toll. Nat Immunol 2008;9:1165–1170.
- Gottar M, Gobert V, Matskevich AA, Reichhart JM, Wang C, Butt TM, Belvin M, Hoffmann JA, Ferrandon D: Dual detection of fungal infections in Drosophila via recognition of glucans and sensing of virulence factors. Cell 2006;127:1425–1437.
- Kambris Z, Brun S, Jang IH, Nam HJ, Romeo Y, Takahashi K, Lee WJ, Ueda R, Lemaitre B: Drosophila immunity: a large-scale in vivo RNAi screen identifies five serine proteases required for Toll activation. Curr Biol 2006;16:808–813.
- Ligoxygakis P, Pelte N, Hoffmann JA, Reichhart JM: Activation of Drosophila Toll during fungal infection by a blood serine protease. Science 2002;297:114–116.
- Rutschmann S, Jung AC, Hetru C, Reichhart JM, Hoffmann JA, Ferrandon D: The Rel protein Dif mediates the antifungal but not the antibacterial host defense in Drosophila. Immunity 2000;12:569–580.
- Busse MS, Arnold CP, Towb P, Katrivesis J, Wasserman SA: A kappaB sequence code for pathway-specific innate immune responses. EMBO J 2007;26:3826–3835.
- Uvell H, Engstrom Y: A multilayered defense against infection: Combinatorial control of insect immune genes. Trends Genet 2007;23:342–349.
- Petersen UM, Kadalayil L, Rehorn KP, Hoshizaki DK, Reuter R, Engstrom Y: Serpent regulates Drosophila immunity genes in the larval fat body through an essential GATA motif. EMBO J 1999;18:4013–4022.
- Senger K, Armstrong GW, Rowell WJ, Kwan JM, Markstein M, Levine M: Immunity regulatory DNAs share common organizational features in Drosophila. Mol Cell 2004;13:19–32.
- Senger K, Harris K, Levine M: GATA factors participate in tissue-specific immune responses in Drosophila larvae. Proc Natl Acad Sci USA 2006;103:15957–15962.
- Ryu JH, Kim SH, Lee HY, Bai JY, Nam YD, Bae JW, Lee DG, Shin SC, Ha EM, Lee WJ: Innate immune homeostasis by the homeobox gene caudal and commensal-gut mutualism in Drosophila. Science 2008;319:777–782.
- Park JM, Kim JM, Kim LK, Kim SN, Kim- Ha J, Kim JH, Kim YJ: Signal-induced transcriptional activation by Dif requires the dTRAP80 mediator module. Mol Cell Biol 2003;23:1358–1367.
- Goto A, Matsushita K, Gesellchen V, El Chamy L, Kuttenkeuler D, Takeuchi O, Hoffmann JA, Akira S, Boutros M, Reichhart JM: Akirins are highly conserved nuclear proteins required for NF-kappaB-dependent gene expression in Drosophila and mice. Nat Immunol 2008;9:97–104.
- Elbashir SM, Lendeckel W, Tuschl T: RNA interference is mediated by 21- and 22-nucleotide RNAs. Genes Dev 2001;15:188–200.
- Fire A, Xu S, Montgomery MK, Kostas SA, Driver SE, Mello CC: Potent and specific genetic interference by double-stranded RNA in Caenorhabditis elegans. Nature 1998;391:806–811.
- Bartscherer K, Pelte N, Ingelfinger D, Boutros M: Secretion of Wnt ligands requires Evi, a conserved transmembrane protein. Cell 2006;125:523–533.
- Gesellchen V, Kuttenkeuler D, Steckel M, Pelte N, Boutros M: An RNA interference screen identifies inhibitor of apoptosis protein 2 as a regulator of innate immune signalling in Drosophila. EMBO Rep 2005;6:979–984.
- Boutros M, Ahringer J: The art and design of genetic screens: RNA interference. Nat Rev 2008;9:554–566.
- Reed DE, Huang XM, Wohlschlegel JA, Levine MS, Senger K: Deaf-1 regulates immunity gene expression in Drosophila. Proc Natl Acad Sci USA 2008;105:8351–8356.
- Boutros M, Kiger AA, Armknecht S, Kerr K, Hild M, Koch B, Haas SA, Paro R, Perrimon N: Genome-wide RNAi analysis of growth and viability in Drosophila cells. Science 2004;303:832–835.
- Sun H, Towb P, Chiem DN, Foster BA, Wasserman SA: Regulated assembly of the Toll signaling complex drives Drosophila dorsoventral patterning. EMBO J 2004;23:100–110.
- Tauszig S, Jouanguy E, Hoffmann JA, Imler JL: Toll-related receptors and the control of antimicrobial peptide expression in Drosophila. Proc Natl Acad Sci USA 2000;97:10520–10525.
- Pfaffl MW: A new mathematical model for relative quantification in real-time RT-PCR. Nucleic Acids Res 2001;29:e45.
- Behm-Ansmant I, Rehwinkel J, Doerks T, Stark A, Bork P, Izaurralde E: mRNA degradation by miRNAs and GW182 requires both CCR4:NOT deadenylase and DCP1:DCP2 decapping complexes. Genes Dev 2006;20:1885–1898.
- Whalen AM, Steward R: Dissociation of the dorsal-cactus complex and phosphorylation of the dorsal protein correlate with the nuclear localization of dorsal. J Cell Biol 1993;123:523–534.
- Lee YS, Carthew RW: Making a better RNAi vector for Drosophila: use of intron spacers. Methods 2003;30:322–329.
- Han ZS, Ip YT: Interaction and specificity of Rel-related proteins in regulating Drosophila immunity gene expression. J Biol Chem 1999;274:21355–21361.
- Tanji T, Hu X, Weber AN, Ip YT: Toll and Imd pathways synergistically activate innate immune response in Drosophila melanogaster. Mol Cell Biol 2007;27:4578–4588.
- Adryan B, Schuh R: Gene-ontology-based clustering of gene expression data. Bioinformatics 2004;20:2851–2852.
- Adryan B, Teichmann SA: Flytf: A systematic review of site-specific transcription factors in the fruit fly Drosophila melanogaster. Bioinformatics 2006;22:1532–1533.
- Boutros M, Bras LP, Huber W: Analysis of cell-based RNAi screens. Genome Biol 2006;7:R66.
- Armstrong JA, Papoulas O, Daubresse G, Sperling AS, Lis JT, Scott MP, Tamkun JW: The Drosophila BRM complex facilitates global transcription by RNA polymerase II. EMBO J 2002;21:5245–5254.
- Horn T, Arziman Z, Berger J, Boutros M: Genome RNAi: a database for cell-based RNAi phenotypes. Nucleic Acids Res 2007;35:D492–D497.
- Arziman Z, Horn T, Boutros M: E-RNAi: a web application to design optimized RNAi constructs. Nucleic Acids Res 2005;33:W582–W588.
- Chintapalli VR, Wang J, Dow JA: Using FlyAtlas to identify better Drosophila melanogaster models of human disease. Nat Genet 2007;39:715–720.
- Gross CT, McGinnis W: Deaf-1, a novel protein that binds an essential region in a deformed response element. EMBO J 1996;15:1961–1970.
- Veraksa A, Kennison J, McGinnis W: Deaf-1 function is essential for the early embryonic development of Drosophila. Genesis 2002;33:67–76.
- Bottomley MJ, Collard MW, Huggenvik JI, Liu Z, Gibson TJ, Sattler M: The SAND domain structure defines a novel DNA-binding fold in transcriptional regulation. Nat Struct Biol 2001;8:626–633.
- Muller P, Kuttenkeuler D, Gesellchen V, Zeidler MP, Boutros M: Identification of JAK/STAT signalling components by genome-wide RNA interference. Nature 2005;436:871–875.
- Hochheimer A, Tjian R: Diversified transcription initiation complexes expand promoter selectivity and tissue-specific gene expression. Genes Dev 2003;17:1309–1320.
- Zeidler MP, Yokomori K, Tjian R, Mlodzik M: Drosophila TFIIA-S is up-regulated and required during Ras-mediated photoreceptor determination. Genes Dev 1996;10:50–59.
- Huggenvik JI, Michelson RJ, Collard MW, Ziemba AJ, Gurley P, Mowen KA: Characterization of a nuclear deformed epidermal autoregulatory factor-1 (Deaf-1)-related (NUDR) transcriptional regulator protein. Mol Endocrinol 1998;12:1619–1639.
- Czesak M, Lemonde S, Peterson EA, Rogaeva A, Albert PR: Cell-specific repressor or enhancer activities of Deaf-1 at a serotonin 1a receptor gene polymorphism. J Neurosci 2006;26:1864–1871.
- Pan H, Yan BS, Rojas M, Shebzukhov YV, Zhou H, Kobzik L, Higgins DE, Daly MJ, Bloom BR, Kramnik I: Ipr1 gene mediates innate immunity to tuberculosis. Nature 2005;434:767–772.
Article / Publication Details
Copyright / Drug Dosage / DisclaimerCopyright: All rights reserved. No part of this publication may be translated into other languages, reproduced or utilized in any form or by any means, electronic or mechanical, including photocopying, recording, microcopying, or by any information storage and retrieval system, without permission in writing from the publisher.
Drug Dosage: The authors and the publisher have exerted every effort to ensure that drug selection and dosage set forth in this text are in accord with current recommendations and practice at the time of publication. However, in view of ongoing research, changes in government regulations, and the constant flow of information relating to drug therapy and drug reactions, the reader is urged to check the package insert for each drug for any changes in indications and dosage and for added warnings and precautions. This is particularly important when the recommended agent is a new and/or infrequently employed drug.
Disclaimer: The statements, opinions and data contained in this publication are solely those of the individual authors and contributors and not of the publishers and the editor(s). The appearance of advertisements or/and product references in the publication is not a warranty, endorsement, or approval of the products or services advertised or of their effectiveness, quality or safety. The publisher and the editor(s) disclaim responsibility for any injury to persons or property resulting from any ideas, methods, instructions or products referred to in the content or advertisements.