title: Manipulation of Cell Cycle and Chromatin Configuration by Means of Cell-Penetrating Geminin.
abstract: Geminin regulates chromatin remodeling and DNA replication licensing which play an important role in regulating cellular proliferation and differentiation. Transcription of the Geminin gene is regulated via an E2F-responsive region, while the protein is being closely regulated by the ubiquitin-proteasome system. Our objective was to directly transduce Geminin protein into cells. Recombinant cell-penetrating Geminin (CP-Geminin) was generated by fusing Geminin with a membrane translocating motif from FGF4 and was efficiently incorporated into NIH 3T3 cells and mouse embryonic fibroblasts. The withdrawal study indicated that incorporated CP-Geminin was quickly reduced after removal from medium. We confirmed CP-Geminin was imported into the nucleus after incorporation and also that the incorporated CP-Geminin directly interacted with Cdt1 or Brahma/Brg1 as the same manner as Geminin. We further demonstrated that incorporated CP-Geminin suppressed S-phase progression of the cell cycle and reduced nuclease accessibility in the chromatin, probably through suppression of chromatin remodeling, indicating that CP-Geminin constitutes a novel tool for controlling chromatin configuration and the cell cycle. Since Geminin has been shown to be involved in regulation of stem cells and cancer cells, CP-Geminin is expected to be useful for elucidating the role of Geminin in stem cells and cancer cells, and for manipulating their activity.
authors.affiliation: Department of Stem Cell Biology, Research Institute for Radiation Biology and Medicine, Hiroshima University, Kasumi, Minami-ku, Hiroshima, Japan.
authors.forename: Yoshinori
authors.initials: Y
authors.lastname: Ohno
authors.affiliation: Department of Stem Cell Biology, Research Institute for Radiation Biology and Medicine, Hiroshima University, Kasumi, Minami-ku, Hiroshima, Japan.
authors.forename: Kyoko
authors.initials: K
authors.lastname: Suzuki-Takedachi
authors.affiliation: Department of Stem Cell Biology, Research Institute for Radiation Biology and Medicine, Hiroshima University, Kasumi, Minami-ku, Hiroshima, Japan.|Department of Biochemistry, Faculty of Medicine, Fukuoka University, Nanakuma, Jonan-ku, Fukuoka, Japan.
authors.forename: Shin'ichiro
authors.initials: S
authors.lastname: Yasunaga
authors.affiliation: Department of Stem Cell Biology, Research Institute for Radiation Biology and Medicine, Hiroshima University, Kasumi, Minami-ku, Hiroshima, Japan.
authors.forename: Toshiaki
authors.initials: T
authors.lastname: Kurogi
authors.affiliation: Department of Stem Cell Biology, Research Institute for Radiation Biology and Medicine, Hiroshima University, Kasumi, Minami-ku, Hiroshima, Japan.
authors.forename: Mimoko
authors.initials: M
authors.lastname: Santo
authors.affiliation: Department of Applied Biological Sciences, College of Bioresource Sciences, Nihon University, Kameino, Fujisawa-city, Kanagawa, Japan.
authors.forename: Yoshikazu
authors.initials: Y
authors.lastname: Masuhiro
authors.affiliation: Department of Applied Biological Sciences, College of Bioresource Sciences, Nihon University, Kameino, Fujisawa-city, Kanagawa, Japan.
authors.forename: Shigemasa
authors.initials: S
authors.lastname: Hanazawa
authors.affiliation: Department of Food and Fermentation Science, Faculty of Food Science and Nutrition, Beppu University, Kita-ishigaki 82, Beppu-city, Oita, Japan.
authors.forename: Motoaki
authors.initials: M
authors.lastname: Ohtsubo
authors.affiliation: Department of Stem Cell Biology, Research Institute for Radiation Biology and Medicine, Hiroshima University, Kasumi, Minami-ku, Hiroshima, Japan.
authors.forename: Kazuhito
authors.initials: K
authors.lastname: Naka
authors.affiliation: Department of Stem Cell Biology, Research Institute for Radiation Biology and Medicine, Hiroshima University, Kasumi, Minami-ku, Hiroshima, Japan.
authors.forename: Yoshihiro
authors.initials: Y
authors.lastname: Takihara
mesh_terms: D020816:Amino Acid Motifs; D000818:Animals; D002453:Cell Cycle; D018797:Cell Cycle Proteins / Q000378:metabolism*; D002843:Chromatin / Q000737:chemistry*; D042002:Chromatin Assembly and Disassembly; D004265:DNA Helicases / Q000378:metabolism*; D004261:DNA Replication; D004268:DNA-Binding Proteins / Q000378:metabolism*; D051519:Fibroblast Growth Factor 4 / Q000378:metabolism; D005347:Fibroblasts / Q000378:metabolism; D064248:Geminin / Q000378:metabolism*; D005786:Gene Expression Regulation; D006801:Humans; D020014:K562 Cells; D051379:Mice; D008810:Mice, Inbred C57BL; D041681:NIH 3T3 Cells; D009687:Nuclear Proteins / Q000378:metabolism*; D010957:Plasmids / Q000378:metabolism; D046988:Proteasome Endopeptidase Complex / Q000737:chemistry; D000067996:RAW 264.7 Cells; D060888:Real-Time Polymerase Chain Reaction; D011994:Recombinant Proteins / Q000378:metabolism; D014157:Transcription Factors / Q000378:metabolism*; D014158:Transcription, Genetic; D014162:Transfection; D025801:Ubiquitin / Q000737:chemistry
chemical_list: D018797:Cell Cycle Proteins; D002843:Chromatin; D004268:DNA-Binding Proteins; C496446:Fgf4 protein, mouse; D051519:Fibroblast Growth Factor 4; D064248:Geminin; C577997:Gmnn protein, mouse; D009687:Nuclear Proteins; D011994:Recombinant Proteins; C498645:Ris2 protein, mouse; D014157:Transcription Factors; D025801:Ubiquitin; D046988:Proteasome Endopeptidase Complex; C513194:Smarca4 protein, mouse; D004265:DNA Helicases