title: Functional Conservation of the Glide/Gcm Regulatory Network Controlling Glia, Hemocyte, and Tendon Cell Differentiation in Drosophila.
abstract: High-throughput screens allow us to understand how transcription factors trigger developmental processes, including cell specification. A major challenge is identification of their binding sites because feedback loops and homeostatic interactions may mask the direct impact of those factors in transcriptome analyses. Moreover, this approach dissects the downstream signaling cascades and facilitates identification of conserved transcriptional programs. Here we show the results and the validation of a DNA adenine methyltransferase identification (DamID) genome-wide screen that identifies the direct targets of Glide/Gcm, a potent transcription factor that controls glia, hemocyte, and tendon cell differentiation in Drosophila. The screen identifies many genes that had not been previously associated with Glide/Gcm and highlights three major signaling pathways interacting with Glide/Gcm: Notch, Hedgehog, and JAK/STAT, which all involve feedback loops. Furthermore, the screen identifies effector molecules that are necessary for cell-cell interactions during late developmental processes and/or in ontogeny. Typically, immunoglobulin (Ig) domain-containing proteins control cell adhesion and axonal navigation. This shows that early and transiently expressed fate determinants not only control other transcription factors that, in turn, implement a specific developmental program but also directly affect late developmental events and cell function. Finally, while the mammalian genome contains two orthologous Gcm genes, their function has been demonstrated in vertebrate-specific tissues, placenta, and parathyroid glands, begging questions on the evolutionary conservation of the Gcm cascade in higher organisms. Here we provide the first evidence for the conservation of Gcm direct targets in humans. In sum, this work uncovers novel aspects of cell specification and sets the basis for further understanding of the role of conserved Gcm gene regulatory cascades.
authors.affiliation: Department of Functional Genomics and Cancer, Institut de Génétique et de Biologie Moléculaire et Cellulaire, UMR7104, F-67404 Illkirch Cedex, France Centre National de la Recherche Scientifique (CNRS), UMR7104, F-67404 Illkirch Cedex, France Institut National de la Santé et de la Recherche Médicale (INSERM), U964, F-67404 Illkirch Cedex, France Université de Strasbourg, F-67404 Illkirch, France.
authors.forename: Pierre B
authors.initials: PB
authors.lastname: Cattenoz
authors.affiliation: Department of Functional Genomics and Cancer, Institut de Génétique et de Biologie Moléculaire et Cellulaire, UMR7104, F-67404 Illkirch Cedex, France Centre National de la Recherche Scientifique (CNRS), UMR7104, F-67404 Illkirch Cedex, France Institut National de la Santé et de la Recherche Médicale (INSERM), U964, F-67404 Illkirch Cedex, France Université de Strasbourg, F-67404 Illkirch, France.
authors.forename: Anna
authors.initials: A
authors.lastname: Popkova
authors.affiliation: Gurdon Institute and Department of Physiology, Development and Neuroscience, University of Cambridge, United Kingdom CB2 1QN.
authors.forename: Tony D
authors.initials: TD
authors.lastname: Southall
authors.affiliation: Department of Functional Genomics and Cancer, Institut de Génétique et de Biologie Moléculaire et Cellulaire, UMR7104, F-67404 Illkirch Cedex, France Centre National de la Recherche Scientifique (CNRS), UMR7104, F-67404 Illkirch Cedex, France Institut National de la Santé et de la Recherche Médicale (INSERM), U964, F-67404 Illkirch Cedex, France Université de Strasbourg, F-67404 Illkirch, France.
authors.forename: Giuseppe
authors.initials: G
authors.lastname: Aiello
authors.affiliation: Gurdon Institute and Department of Physiology, Development and Neuroscience, University of Cambridge, United Kingdom CB2 1QN.
authors.forename: Andrea H
authors.initials: AH
authors.lastname: Brand
authors.affiliation: Department of Functional Genomics and Cancer, Institut de Génétique et de Biologie Moléculaire et Cellulaire, UMR7104, F-67404 Illkirch Cedex, France Centre National de la Recherche Scientifique (CNRS), UMR7104, F-67404 Illkirch Cedex, France Institut National de la Santé et de la Recherche Médicale (INSERM), U964, F-67404 Illkirch Cedex, France Université de Strasbourg, F-67404 Illkirch, France angela@igbmc.fr.
authors.forename: Angela
authors.initials: A
authors.lastname: Giangrande
mesh_terms: D000818:Animals; D001665:Binding Sites; D002454:Cell Differentiation; D002460:Cell Line; D017124:Conserved Sequence; D004247:DNA / Q000378:metabolism; D004268:DNA-Binding Proteins / Q000378:metabolism / Q000502:physiology*; D004330:Drosophila / Q000166:cytology*; D029721:Drosophila Proteins / Q000378:metabolism / Q000502:physiology*; D025461:Feedback, Physiological; D005821:Genetic Techniques; D006367:HeLa Cells; D006434:Hemocytes / Q000166:cytology; D006801:Humans; D009457:Neuroglia / Q000166:cytology; D015398:Signal Transduction; D015265:Site-Specific DNA-Methyltransferase (Adenine-Specific) / Q000378:metabolism; D014157:Transcription Factors / Q000378:metabolism / Q000502:physiology*
chemical_list: D004268:DNA-Binding Proteins; D029721:Drosophila Proteins; C102805:GCM protein, Drosophila; D014157:Transcription Factors; D004247:DNA; D015265:Site-Specific DNA-Methyltransferase (Adenine-Specific)
keywords: DamID; Drosophila; glide/gcm; mGcm; screen