DEAF-1 regulates immunity gene expression in Drosophila.
Immunity genes are activated in the Drosophila fat body by Rel and GATA transcription factors. Here, we present evidence that an additional regulatory factor, deformed epidermal autoregulatory factor-1 (DEAF-1), also contributes to the immune response and is specifically important for the induction of two genes encoding antimicrobial peptides, Metchnikowin (Mtk) and Drosomycin (Drs). The systematic mutagenesis of a minimal Mtk 5' enhancer identified a sequence motif essential for both a response to LPS preparations in S2 cells and activation in the larval fat body in response to bacterial infection. Using affinity chromatography coupled to multidimensional protein identification technology (MudPIT), we identified DEAF-1 as a candidate regulator. DEAF-1 activates the expression of Mtk and Drs promoter-luciferase fusion genes in S2 cells. SELEX assays and footprinting data indicate that DEAF-1 binds to and activates Mtk and Drs regulatory DNAs via a TTCGGBT motif. The insertion of this motif into the Diptericin (Dpt) regulatory region confers DEAF-1 responsiveness to this normally DEAF-1-independent enhancer. The coexpression of DEAF-1 with Dorsal, Dif, and Relish results in the synergistic activation of transcription. We propose that DEAF-1 is a regulator of Drosophila immunity.
Reed
Darien E
DE
Helen Diller Family Comprehensive Cancer Center and Cancer Research Institute, University of California, San Francisco, CA 94115-0128, USA.
Huang
Xinhua M
XM
Wohlschlegel
James A
JA
Levine
Michael S
MS
Senger
Kate
K
D000818:Animals; D023181:Antimicrobial Cationic Peptides / Q000235:genetics*; D001483:Base Sequence; D004268:DNA-Binding Proteins / Q000235:genetics; D029721:Drosophila Proteins / Q000235:genetics* / Q000378:metabolism*; D004331:Drosophila melanogaster / Q000235:genetics / Q000276:immunology* / Q000382:microbiology; D004742:Enhancer Elements, Genetic; D050939:Gene Fusion; D017930:Genes, Reporter; D007109:Immunity / Q000235:genetics*; D008070:Lipopolysaccharides / Q000276:immunology; D008156:Luciferases / Q000235:genetics; D008969:Molecular Sequence Data; D016296:Mutagenesis; D009154:Mutation; D009687:Nuclear Proteins / Q000235:genetics / Q000378:metabolism*; D010750:Phosphoproteins / Q000235:genetics; D011401:Promoter Regions, Genetic; D014157:Transcription Factors / Q000235:genetics; D015533:Transcriptional Activation
D023181:Antimicrobial Cationic Peptides; D004268:DNA-Binding Proteins; C485477:Deaf1 protein, Drosophila; C084428:Dif protein, Drosophila; D029721:Drosophila Proteins; D008070:Lipopolysaccharides; D009687:Nuclear Proteins; D010750:Phosphoproteins; C101626:Rel protein, Drosophila; D014157:Transcription Factors; C062236:dl protein, Drosophila; C091227:DRS protein, Drosophila; C096696:Mtk protein, Drosophila; D008156:Luciferases