title: EVEN-SKIPPED HOMEOBOX 1 controls human ES cell differentiation by directly repressing GOOSECOID expression.
abstract: TGFß signaling patterns the primitive streak, yet little is known about transcriptional effectors that mediate the cell fate choices during streak-like development in mammalian embryos and in embryonic stem (ES) cells. Here we demonstrate that cross-antagonistic actions of EVEN-SKIPPED HOMEOBOX 1 (EVX1) and GOOSECOID (GSC) regulate cell fate decisions in streak-like progenitors derived from human ES cells exposed to BMP4 and/or activin. We found that EVX1 repressed GSC expression and promoted formation of posterior streak-like progeny in response to BMP4, and conversely that GSC repressed EVX1 expression and was required for development of anterior streak-like progeny in response to activin. Chromatin immunoprecipitation assays showed that EVX1 bound to the GSC 5'-flanking region in BMP4 treated human ES cells, and band shift assays identified two EVX1 binding sites in the GSC 5'-region. Significantly, we found that intact EVX1 binding sites were required for BMP4-mediated repression of GSC reporter constructs. We conclude that BMP4-induced EVX1 repress GSC directly and the two genes form the core of a gene regulatory network (GRN) controlling cell fates in streak-like human ES cell progeny.
authors.affiliation: Department of Developmental Biology, Hagedorn Research Institute, Gentofte, Denmark. pas@hagedorn.dk
authors.forename: Mark
authors.initials: M
authors.lastname: Kalisz
authors.forename: Maria
authors.initials: M
authors.lastname: Winzi
authors.forename: Hanne Cathrine
authors.initials: HC
authors.lastname: Bisgaard
authors.forename: Palle
authors.initials: P
authors.lastname: Serup
mesh_terms: D028341:Activins / Q000378:metabolism; D000704:Analysis of Variance; D001665:Binding Sites / Q000235:genetics; D015153:Blotting, Western; D055415:Bone Morphogenetic Protein 4 / Q000378:metabolism; D002454:Cell Differentiation / Q000502:physiology*; D047369:Chromatin Immunoprecipitation; D017931:DNA Primers / Q000235:genetics; D024202:Electrophoretic Mobility Shift Assay; D053595:Embryonic Stem Cells / Q000502:physiology*; D005434:Flow Cytometry; D018507:Gene Expression Regulation, Developmental / Q000235:genetics / Q000502:physiology*; D055785:Gene Knockdown Techniques; D053263:Gene Regulatory Networks / Q000235:genetics*; D051956:Goosecoid Protein / Q000235:genetics / Q000378:metabolism*; D018398:Homeodomain Proteins / Q000235:genetics / Q000378:metabolism*; D006801:Humans; D007150:Immunohistochemistry; D008156:Luciferases; D054240:Primitive Streak / Q000166:cytology / Q000196:embryology*; D060888:Real-Time Polymerase Chain Reaction; D020133:Reverse Transcriptase Polymerase Chain Reaction
chemical_list: C525809:BMP4 protein, human; D055415:Bone Morphogenetic Protein 4; D017931:DNA Primers; C497992:GSC protein, human; D051956:Goosecoid Protein; D018398:Homeodomain Proteins; D028341:Activins; C074437:EVX1 protein, human; D008156:Luciferases