title: ETS-related transcription factors ETV4 and ETV5 are involved in proliferation and induction of differentiation-associated genes in embryonic stem (ES) cells.
abstract: The pluripotency and self-renewal capacity of embryonic stem (ES) cells is regulated by several transcription factors. Here, we show that the ETS-related transcription factors Etv4 and Etv5 (Etv4/5) are specifically expressed in undifferentiated ES cells, and suppression of Oct3/4 results in down-regulation of Etv4/5. Simultaneous deletion of Etv4 and Etv5 (Etv4/5 double knock-out (dKO)) in ES cells resulted in a flat, epithelial cell-like appearance, whereas the morphology changed into compact colonies in a 2i medium (containing two inhibitors for GSK3 and MEK/ERK). Expression levels of self-renewal marker genes, including Oct3/4 and Nanog, were similar between wild-type and dKO ES cells, whereas proliferation of Etv4/5 dKO ES cells was decreased with overexpression of cyclin-dependent kinase inhibitors (p16/p19, p15, and p57). A differentiation assay revealed that the embryoid bodies derived from Etv4/5 dKO ES cells were smaller than the control, and expression of ectoderm marker genes, including Fgf5, Sox1, and Pax3, was not induced in dKO-derived embryoid bodies. Microarray analysis demonstrated that stem cell-related genes, including Tcf15, Gbx2, Lrh1, Zic3, and Baf60c, were significantly repressed in Etv4/5 dKO ES cells. The artificial expression of Etv4 and/or Etv5 in Etv4/5 dKO ES cells induced re-expression of Tcf15 and Gbx2. These results indicate that Etv4 and Etv5, potentially through regulation of Gbx2 and Tcf15, are involved in the ES cell proliferation and induction of differentiation-associated genes in ES cells.
authors.affiliation: From the Department of Stem Cell Biology, Graduate School of Medical Sciences, Kanazawa University, 13-1 Takara-machi, Kanazawa, Ishikawa 920-8640, Japan, tadayuki@staff.kanazawa-u.ac.jp.
authors.forename: Tadayuki
authors.initials: T
authors.lastname: Akagi
authors.affiliation: the Institute of Biotechnology, University of Helsinki, 00790 Helsinki, Finland, and.
authors.forename: Satu
authors.initials: S
authors.lastname: Kuure
authors.affiliation: From the Department of Stem Cell Biology, Graduate School of Medical Sciences, Kanazawa University, 13-1 Takara-machi, Kanazawa, Ishikawa 920-8640, Japan.
authors.forename: Kousuke
authors.initials: K
authors.lastname: Uranishi
authors.affiliation: From the Department of Stem Cell Biology, Graduate School of Medical Sciences, Kanazawa University, 13-1 Takara-machi, Kanazawa, Ishikawa 920-8640, Japan.
authors.forename: Hiroshi
authors.initials: H
authors.lastname: Koide
authors.affiliation: the Department of Genetics and Development, Columbia University Medical Center, New York, New York 10032.
authors.forename: Frank
authors.initials: F
authors.lastname: Costantini
authors.affiliation: From the Department of Stem Cell Biology, Graduate School of Medical Sciences, Kanazawa University, 13-1 Takara-machi, Kanazawa, Ishikawa 920-8640, Japan, tyokota@med.kanazawa-u.ac.jp.
authors.forename: Takashi
authors.initials: T
authors.lastname: Yokota
mesh_terms: D000818:Animals; D000943:Antigens, Differentiation / Q000096:biosynthesis / Q000235:genetics; D002454:Cell Differentiation / Q000502:physiology*; D049109:Cell Proliferation / Q000502:physiology*; D050756:Cyclin-Dependent Kinase Inhibitor Proteins / Q000096:biosynthesis / Q000235:genetics; D004268:DNA-Binding Proteins / Q000235:genetics / Q000378:metabolism*; D053595:Embryonic Stem Cells / Q000166:cytology / Q000378:metabolism*; D038362:Glycogen Synthase Kinase 3 / Q000096:biosynthesis / Q000235:genetics; D051379:Mice; D018345:Mice, Knockout; D050783:Proto-Oncogene Proteins c-ets / Q000235:genetics / Q000378:metabolism*; D014157:Transcription Factors / Q000235:genetics / Q000378:metabolism*
chemical_list: D000943:Antigens, Differentiation; D050756:Cyclin-Dependent Kinase Inhibitor Proteins; D004268:DNA-Binding Proteins; C540571:Etv4 protein, mouse; C513242:Etv5 protein, mouse; D050783:Proto-Oncogene Proteins c-ets; D014157:Transcription Factors; D038362:Glycogen Synthase Kinase 3
keywords: cell proliferation; differentiation; embryonic stem cell; gene regulation; oncogene; transcription factor