title: Tissue-specific enhancer repression through molecular integration of cell signaling inputs.
abstract: Drosophila leg morphogenesis occurs under the control of a relatively well-known genetic cascade, which mobilizes both cell signaling pathways and tissue-specific transcription factors. However, their cross-regulatory interactions, deployed to refine leg patterning, remain poorly characterized at the gene expression level. Within the genetically interacting landscape that governs limb development, the bric-à-brac2 (bab2) gene is required for distal leg segmentation. We have previously shown that the Distal-less (Dll) homeodomain and Rotund (Rn) zinc-finger activating transcription factors control limb-specific bab2 expression by binding directly a single critical leg/antennal enhancer (LAE) within the bric-à-brac locus. By genetic and molecular analyses, we show here that the EGFR-responsive C15 homeodomain and the Notch-regulated Bowl zinc-finger transcription factors also interact directly with the LAE enhancer as a repressive duo. The appendage patterning gene bab2 is the first identified direct target of the Bowl repressor, an Odd-skipped/Osr family member. Moreover, we show that C15 acts on LAE activity independently of its regular partner, the Aristaless homeoprotein. Instead, we find that C15 interacts physically with the Dll activator through contacts between their homeodomain and binds competitively with Dll to adjacent cognate sites on LAE, adding potential new layers of regulation by C15. Lastly, we show that C15 and Bowl activities regulate also rn expression. Our findings shed light on how the concerted action of two transcriptional repressors, in response to cell signaling inputs, shapes and refines gene expression along the limb proximo-distal axis in a timely manner.
authors.affiliation: Centre de Biologie du Développement (CBD), Centre de Biologie Intégrative (CBI), Université Fédérale de Toulouse, UMR5547 CNRS/Université Paul Sabatier (UPS), Toulouse, France.
authors.forename: Luis Humberto
authors.identifier: 0000-0002-5654-8265
authors.initials: LH
authors.lastname: Mojica-Vázquez
authors.affiliation: Centre de Biologie du Développement (CBD), Centre de Biologie Intégrative (CBI), Université Fédérale de Toulouse, UMR5547 CNRS/Université Paul Sabatier (UPS), Toulouse, France.
authors.forename: Mikhail H
authors.initials: MH
authors.lastname: Benetah
authors.affiliation: Centre de Biologie du Développement (CBD), Centre de Biologie Intégrative (CBI), Université Fédérale de Toulouse, UMR5547 CNRS/Université Paul Sabatier (UPS), Toulouse, France.
authors.forename: Aissette
authors.initials: A
authors.lastname: Baanannou
authors.affiliation: Centre de Biologie du Développement (CBD), Centre de Biologie Intégrative (CBI), Université Fédérale de Toulouse, UMR5547 CNRS/Université Paul Sabatier (UPS), Toulouse, France.
authors.forename: Sandra
authors.initials: S
authors.lastname: Bernat-Fabre
authors.affiliation: Laboratory of Systems Biology and Genetics, Institute of Bioengineering, School of Life Sciences, École Polytechnique Fédérale de Lausanne (EPFL), Lausanne, Switzerland.
authors.forename: Bart
authors.identifier: 0000-0001-9935-843X
authors.initials: B
authors.lastname: Deplancke
authors.affiliation: Centre de Biologie du Développement (CBD), Centre de Biologie Intégrative (CBI), Université Fédérale de Toulouse, UMR5547 CNRS/Université Paul Sabatier (UPS), Toulouse, France.
authors.forename: David L
authors.initials: DL
authors.lastname: Cribbs
authors.affiliation: Centre de Biologie du Développement (CBD), Centre de Biologie Intégrative (CBI), Université Fédérale de Toulouse, UMR5547 CNRS/Université Paul Sabatier (UPS), Toulouse, France.
authors.forename: Henri-Marc
authors.identifier: 0000-0001-9901-2260
authors.initials: HM
authors.lastname: Bourbon
authors.affiliation: Centre de Biologie du Développement (CBD), Centre de Biologie Intégrative (CBI), Université Fédérale de Toulouse, UMR5547 CNRS/Université Paul Sabatier (UPS), Toulouse, France.
authors.forename: Muriel
authors.identifier: 0000-0002-4444-2519
authors.initials: M
authors.lastname: Boube
mesh_terms: D000818:Animals; D001665:Binding Sites; D004268:DNA-Binding Proteins / Q000096:biosynthesis / Q000235:genetics*; D029721:Drosophila Proteins / Q000096:biosynthesis / Q000235:genetics* / Q000378:metabolism; D004331:Drosophila melanogaster / Q000235:genetics* / Q000254:growth & development; D010450:Endopeptidases / Q000096:biosynthesis / Q000235:genetics*; D004742:Enhancer Elements, Genetic; D066246:ErbB Receptors / Q000235:genetics; D005121:Extremities / Q000254:growth & development; D018398:Homeodomain Proteins / Q000235:genetics* / Q000378:metabolism; D009024:Morphogenesis / Q000235:genetics*; D009928:Organ Specificity / Q000235:genetics; D011485:Protein Binding; D018010:Receptors, Invertebrate Peptide / Q000235:genetics; D012097:Repressor Proteins / Q000096:biosynthesis / Q000235:genetics*; D015398:Signal Transduction; D014157:Transcription Factors / Q000096:biosynthesis / Q000235:genetics* / Q000378:metabolism
chemical_list: C459600:BAB2 protein, Drosophila; C499642:C15 protein, Drosophila; D004268:DNA-Binding Proteins; D029721:Drosophila Proteins; D018398:Homeodomain Proteins; D018010:Receptors, Invertebrate Peptide; D012097:Repressor Proteins; D014157:Transcription Factors; C100532:bowl protein, Drosophila; C117261:distal-less protein, insect; C066532:Egfr protein, Drosophila; D066246:ErbB Receptors; D010450:Endopeptidases; C499246:RN-tre protein, Drosophila