title: Activation of Ftz-F1-Responsive Genes through Ftz/Ftz-F1 Dependent Enhancers.
abstract: The orphan nuclear receptor Ftz-F1 is expressed in all somatic nuclei in Drosophila embryos, but mutations result in a pair-rule phenotype. This was explained by the interaction of Ftz-F1 with the homeodomain protein Ftz that is expressed in stripes in the primordia of segments missing in either ftz-f1 or ftz mutants. Ftz-F1 and Ftz were shown to physically interact and coordinately activate the expression of ftz itself and engrailed by synergistic binding to composite Ftz-F1/Ftz binding sites. However, attempts to identify additional target genes on the basis of Ftz-F1/ Ftz binding alone has met with only limited success. To discern rules for Ftz-F1 target site selection in vivo and to identify additional target genes, a microarray analysis was performed comparing wildtype and ftz-f1 mutant embryos. Ftz-F1-responsive genes most highly regulated included engrailed and nine additional genes expressed in patterns dependent on both ftz and ftz-f1. Candidate enhancers for these genes were identified by combining BDTNP Ftz ChIP-chip data with a computational search for Ftz-F1 binding sites. Of eight enhancer reporter genes tested in transgenic embryos, six generated expression patterns similar to the corresponding endogenous gene and expression was lost in ftz mutants. These studies identified a new set of Ftz-F1 targets, all of which are co-regulated by Ftz. Comparative analysis of enhancers containing Ftz/Ftz-F1 binding sites that were or were not bona fide targets in vivo suggested that GAF negatively regulates enhancers that contain Ftz/Ftz-F1 binding sites but are not actually utilized. These targets include other regulatory factors as well as genes involved directly in morphogenesis, providing insight into how pair-rule genes establish the body pattern.
authors.affiliation: Department of Entomology and Program in Molecular & Cell Biology, University of Maryland, College Park, Maryland, 20742, United States of America.
authors.forename: Amanda
authors.initials: A
authors.lastname: Field
authors.affiliation: Department of Entomology and Program in Molecular & Cell Biology, University of Maryland, College Park, Maryland, 20742, United States of America.
authors.forename: Jie
authors.initials: J
authors.lastname: Xiang
authors.affiliation: Department of Entomology and Program in Molecular & Cell Biology, University of Maryland, College Park, Maryland, 20742, United States of America.
authors.forename: W Ray
authors.initials: WR
authors.lastname: Anderson
authors.affiliation: Department of Entomology and Program in Molecular & Cell Biology, University of Maryland, College Park, Maryland, 20742, United States of America.
authors.forename: Patricia
authors.initials: P
authors.lastname: Graham
authors.affiliation: Department of Entomology and Program in Molecular & Cell Biology, University of Maryland, College Park, Maryland, 20742, United States of America.
authors.forename: Leslie
authors.initials: L
authors.lastname: Pick
mesh_terms: D000818:Animals; D004268:DNA-Binding Proteins / Q000235:genetics / Q000378:metabolism*; D029721:Drosophila Proteins / Q000235:genetics / Q000378:metabolism*; D004331:Drosophila melanogaster; D051717:Fushi Tarazu Transcription Factors / Q000235:genetics / Q000378:metabolism*; D018507:Gene Expression Regulation, Developmental / Q000502:physiology*; D009024:Morphogenesis / Q000502:physiology*; D020218:Response Elements / Q000502:physiology*; D014157:Transcription Factors / Q000235:genetics / Q000378:metabolism*
chemical_list: D004268:DNA-Binding Proteins; D029721:Drosophila Proteins; D051717:Fushi Tarazu Transcription Factors; D014157:Transcription Factors; C497169:ftz protein, Drosophila; C499712:nuclear hormone receptor FTZ-F1, Drosophila