title: Comparative gene expression analysis of Dtg, a novel target gene of Dpp signaling pathway in the early Drosophila melanogaster embryo.
abstract: In the early Drosophila melanogaster embryo, Dpp, a secreted molecule that belongs to the TGF-β superfamily of growth factors, activates a set of downstream genes to subdivide the dorsal region into amnioserosa and dorsal epidermis. Here, we examined the expression pattern and transcriptional regulation of Dtg, a new target gene of Dpp signaling pathway that is required for proper amnioserosa differentiation. We showed that the expression of Dtg was controlled by Dpp and characterized a 524-bp enhancer that mediated expression in the dorsal midline, as well as, in the differentiated amnioserosa in transgenic reporter embryos. This enhancer contained a highly conserved region of 48-bp in which bioinformatic predictions and in vitro assays identified three Mad binding motifs. Mutational analysis revealed that these three motifs were necessary for proper expression of a reporter gene in transgenic embryos, suggesting that short and highly conserved genomic sequences may be indicative of functional regulatory regions in D. melanogaster genes. Dtg orthologs were not detected in basal lineages of Dipterans, which unlike D. melanogaster develop two extra-embryonic membranes, amnion and serosa, nevertheless Dtg orthologs were identified in the transcriptome of Musca domestica, in which dorsal ectoderm patterning leads to the formation of a single extra-embryonic membrane. These results suggest that Dtg was recruited as a new component of the network that controls dorsal ectoderm patterning in the lineage leading to higher Cyclorrhaphan flies, such as D. melanogaster and M. domestica.
authors: Laboratorio de Bioinformática y Expresión Génica, INTA-Universidad de Chile, El Líbano 5524, Santiago, Chile; Fondap Center for Genome Regulation (CGR), Universidad de Chile, Santiago, Chile.
authors: Christian
authors: C
authors: Hodar
authors: Laboratorio de Bioinformática y Expresión Génica, INTA-Universidad de Chile, El Líbano 5524, Santiago, Chile; Fondap Center for Genome Regulation (CGR), Universidad de Chile, Santiago, Chile.
authors: Alejandro
authors: A
authors: Zuñiga
authors: Laboratorio de Bioinformática y Expresión Génica, INTA-Universidad de Chile, El Líbano 5524, Santiago, Chile; Fondap Center for Genome Regulation (CGR), Universidad de Chile, Santiago, Chile.
authors: Rodrigo
authors: R
authors: Pulgar
authors: Laboratorio de Bioinformática y Matemática del Genoma, Center for Mathematical Modeling, FCFM-Universidad de Chile, Santiago, Chile; Fondap Center for Genome Regulation (CGR), Universidad de Chile, Santiago, Chile.
authors: Dante
authors: D
authors: Travisany
authors: Laboratorio de Bioinformática y Expresión Génica, INTA-Universidad de Chile, El Líbano 5524, Santiago, Chile.
authors: Carlos
authors: C
authors: Chacon
authors: Laboratorio de Bioinformática y Expresión Génica, INTA-Universidad de Chile, El Líbano 5524, Santiago, Chile.
authors: Michael
authors: M
authors: Pino
authors: Laboratorio de Bioinformática y Matemática del Genoma, Center for Mathematical Modeling, FCFM-Universidad de Chile, Santiago, Chile; Fondap Center for Genome Regulation (CGR), Universidad de Chile, Santiago, Chile; Department of Mathematical Engineering, FCFM-Universidad de Chile, Santiago, Chile.
authors: Alejandro
authors: A
authors: Maass
authors: Laboratorio de Bioinformática y Expresión Génica, INTA-Universidad de Chile, El Líbano 5524, Santiago, Chile; Fondap Center for Genome Regulation (CGR), Universidad de Chile, Santiago, Chile. Electronic address: vcambiaz@inta.uchile.cl.
authors: Verónica
authors: V
authors: Cambiazo
mesh_terms: D000818:Animals; D001483:Base Sequence; D029721:Drosophila Proteins / Q000235:genetics* / Q000378:metabolism; D004331:Drosophila melanogaster / Q000201:enzymology / Q000235:genetics* / Q000378:metabolism*; D004625:Embryo, Nonmammalian; D004742:Enhancer Elements, Genetic; D020869:Gene Expression Profiling; D018507:Gene Expression Regulation, Developmental; D008565:Membrane Proteins / Q000235:genetics*; D011485:Protein Binding; D016415:Sequence Alignment; D015398:Signal Transduction; D013045:Species Specificity
chemical_list: D029721:Drosophila Proteins; C586900:Dtg protein, Drosophila; D008565:Membrane Proteins; C064196:dpp protein, Drosophila
keywords: 4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid; Amnioserosa; BMP; Bone morphogenetic protein; Copper sulfate; CuSO4; Cyclorrhaphan flies; D. ananassae; D. erecta; D. grimshawi; D. melanogaster; D. mojavensis; D. persimilis; D. pseudoobscura; D. sechelia; D. simulans; D. virilis; D. willistoni; D. yakuba; DIG; DTT; DV; Decapentaplegic; Digoxigenin; Dithiothreitol; Dorsal target gene; Dorso ventral; Dpp; Drosophila ananassae; Drosophila erecta; Drosophila grimshawi; Drosophila melanogaster; Drosophila mojavensis; Drosophila persimilis; Drosophila pseudoobscura; Drosophila sechelia; Drosophila simulans; Drosophila virilis; Drosophila willistoni; Drosophila yakuba; Dtg; EDTA; EGTA; Ethylene glycol tetraacetic acid; Ethylenediaminetetraacetic acid; FITC; Fluorescein isothiocyanate; GAL4; GST; Glutathione S-transferase; HCl; HEPES; Hydrochloric acid; KCl; KOAc; LB; Luria–Bertani (medium); M. domestica; MAST; MEME; MH1; MLAGAN; MUSCLE; Mad; Mad homology 1 domain; Magnesium chloride; Messenger ribonucleic acid; MgCl(2); Millions years ago; Mothers against decapentaplegic; Motif Alignment and Search Tool; Multiple em (expectation maximization) for motif elicitation; Multiple limited area global alignment of nucleotides; Multiple sequence comparison by log-expectation; Musca domestica; Mya; NaCl; NaOCl; PBS; PCR; PEG; Phosphate buffered saline; Phosphorylated MAD; Polyethylene glycol; Polymerase chain reaction; Potassium acetate; Potassium chloride; SDS-PAGE; Screw; Scw; Short gastrulation; Sodium chloride; Sodium dodecyl sulfate PA-gel electrophoresis; Sodium hypochlorite; Sog; TGF-β; TMHMM; Trans membrane hidden Markov model; Transforming growth factor beta; Tris; UAS; Upstream activation sequence; Yeast transcription activator protein; Zinc sulfate; Zn2SO4; base pair; bp; kb; kilobase; lacZ; mRNA; pMad; tris(hydroxymethyl)aminomethane; β-galactosidase gen