Genome-wide identification of in vivo Drosophila Engrailed-binding DNA fragments and related target genes.
Chromatin immunoprecipitation after UV crosslinking of DNA/protein interactions was used to construct a library enriched in genomic sequences that bind to the Engrailed transcription factor in Drosophila embryos. Sequencing of the clones led to the identification of 203 Engrailed-binding fragments localized in intergenic or intronic regions. Genes lying near these fragments, which are considered as potential Engrailed target genes, are involved in different developmental pathways, such as anteroposterior patterning, muscle development, tracheal pathfinding or axon guidance. We validated this approach by in vitro and in vivo tests performed on a subset of Engrailed potential targets involved in these various pathways. Finally, we present strong evidence showing that an immunoprecipitated genomic DNA fragment corresponds to a promoter region involved in the direct regulation of frizzled2 expression by engrailed in vivo.
Solano
Pascal Jean
PJ
Institut de Génétique Humaine (UPR 1142). 141 rue de la Cardonille, 34396 Montpellier, France.
Mugat
Bruno
B
Martin
David
D
Girard
Franck
F
Huibant
Jean-Marc
JM
Ferraz
Conchita
C
Jacq
Bernard
B
Demaille
Jacques
J
Maschat
Florence
F
D000818:Animals; D004247:DNA / Q000378:metabolism*; D004330:Drosophila / Q000235:genetics* / Q000378:metabolism*; D024202:Electrophoretic Mobility Shift Assay; D005786:Gene Expression Regulation; D018398:Homeodomain Proteins / Q000378:metabolism*; D007814:Larva / Q000378:metabolism; D011956:Receptors, Cell Surface / Q000378:metabolism; D014157:Transcription Factors
D018398:Homeodomain Proteins; D011956:Receptors, Cell Surface; D014157:Transcription Factors; C106185:engrailed homeobox proteins; D004247:DNA