title: Direct transcriptional induction of Gadd45gamma by Ascl1 during neuronal differentiation.
abstract: The basic helix-loop-helix transcription factor Ascl1 plays a critical role in the intrinsic genetic program responsible for neuronal differentiation. Here, we describe a novel model system of P19 embryonic carcinoma cells with doxycycline-inducible expression of Ascl1. Microarray hybridization and real-time PCR showed that these cells demonstrated increased expression of many neuronal proteins in a time- and concentration-dependent manner. Interestingly, the gene encoding the cell cycle regulator Gadd45gamma was increased earliest and to the greatest extent following Ascl1 induction. Here, we provide the first evidence identifying Gadd45gamma as a direct transcriptional target of Ascl1. Transactivation and chromatin immunoprecipitation assays identified two E-box consensus sites within the Gadd45gamma promoter necessary for Ascl1 regulation, and demonstrated that Ascl1 is bound to this region within the Gadd45gamma promoter. Furthermore, we found that overexpression of Gadd45gamma itself is sufficient to initiate some aspects of neuronal differentiation independent of Ascl1.
authors.affiliation: Neuroscience Graduate Program, University of Michigan, Ann Arbor, MI 48105, USA.
authors.forename: Holly S
authors.initials: HS
authors.lastname: Huang
authors.forename: Ginger M
authors.initials: GM
authors.lastname: Kubish
authors.forename: Tanya M
authors.initials: TM
authors.lastname: Redmond
authors.forename: David L
authors.initials: DL
authors.lastname: Turner
authors.forename: Robert C
authors.initials: RC
authors.lastname: Thompson
authors.forename: Geoffrey G
authors.initials: GG
authors.lastname: Murphy
authors.forename: Michael D
authors.initials: MD
authors.lastname: Uhler
mesh_terms: D000818:Animals; D000900:Anti-Bacterial Agents / Q000494:pharmacology; D051792:Basic Helix-Loop-Helix Proteins / Q000235:genetics / Q000378:metabolism*; D002454:Cell Differentiation / Q000187:drug effects / Q000502:physiology*; D004318:Doxycycline / Q000494:pharmacology; D005786:Gene Expression Regulation / Q000187:drug effects; D018257:Helix-Loop-Helix Motifs; D006801:Humans; D047908:Intracellular Signaling Peptides and Proteins / Q000235:genetics / Q000378:metabolism*; D051379:Mice; D046228:Microarray Analysis; D009474:Neurons / Q000166:cytology / Q000187:drug effects / Q000502:physiology*; D018408:Patch-Clamp Techniques; D011401:Promoter Regions, Genetic; D014158:Transcription, Genetic; D014407:Tumor Cells, Cultured; D000097642:GADD45 Proteins
chemical_list: D000900:Anti-Bacterial Agents; D051792:Basic Helix-Loop-Helix Proteins; D004318:Doxycycline; D047908:Intracellular Signaling Peptides and Proteins; D000097642:GADD45 Proteins; C499317:Ascl1 protein, mouse
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