title: Octamer and Sox elements are required for transcriptional cis regulation of Nanog gene expression.
abstract: The pluripotential cell-specific gene Nanog encodes a homeodomain-bearing transcription factor required for maintaining the undifferentiated state of stem cells. However, the molecular mechanisms that regulate Nanog gene expression are largely unknown. To address this important issue, we used luciferase assays to monitor the relative activities of deletion fragments from the 5'-flanking region of the gene. An adjacent pair of highly conserved Octamer- and Sox-binding sites was found to be essential for activating pluripotential state-specific gene expression. Furthermore, the 5'-end fragment encompassing the Octamer/Sox element was sufficient for inducing the proper expression of a green fluorescent protein reporter gene even in human embryonic stem (ES) cells. The potential of OCT4 and SOX2 to bind to this element was verified by electrophoretic mobility shift assays with extracts from F9 embryonal carcinoma cells and embryonic germ cells derived from embryonic day 12.5 embryos. However, in ES cell extracts, a complex of OCT4 with an undefined factor preferentially bound to the Octamer/Sox element. Thus, Nanog transcription may be regulated through an interaction between Oct4 and Sox2 or a novel pluripotential cell-specific Sox element-binding factor which is prominent in ES cells.
authors.affiliation: Department of Development and Differentiation, Institute for Frontier Medical Sciences, Kyoto University, Shogoin, Sakyo-ku, Kyoto, Japan.
authors.forename: Takao
authors.initials: T
authors.lastname: Kuroda
authors.forename: Masako
authors.initials: M
authors.lastname: Tada
authors.forename: Hiroshi
authors.initials: H
authors.lastname: Kubota
authors.forename: Hironobu
authors.initials: H
authors.lastname: Kimura
authors.forename: Shin-ya
authors.initials: SY
authors.lastname: Hatano
authors.forename: Hirofumi
authors.initials: H
authors.lastname: Suemori
authors.forename: Norio
authors.initials: N
authors.lastname: Nakatsuji
authors.forename: Takashi
authors.initials: T
authors.lastname: Tada
mesh_terms: D024506:5' Flanking Region / Q000235:genetics; D000818:Animals; D001483:Base Sequence; D001665:Binding Sites / Q000235:genetics; D004268:DNA-Binding Proteins / Q000235:genetics* / Q000378:metabolism / Q000502:physiology*; D024202:Electrophoretic Mobility Shift Assay; D004622:Embryo, Mammalian / Q000166:cytology; D005786:Gene Expression Regulation; D017930:Genes, Reporter / Q000235:genetics; D049452:Green Fluorescent Proteins / Q000032:analysis / Q000235:genetics; D024002:HMGB Proteins; D018398:Homeodomain Proteins / Q000235:genetics*; D006801:Humans; D008156:Luciferases / Q000032:analysis / Q000235:genetics; D051379:Mice; D008969:Molecular Sequence Data; D009154:Mutation / Q000235:genetics; D000071317:Nanog Homeobox Protein; D009687:Nuclear Proteins / Q000235:genetics / Q000378:metabolism / Q000502:physiology*; D050814:Octamer Transcription Factor-3; D020218:Response Elements / Q000235:genetics*; D055748:SOXB1 Transcription Factors; D017384:Sequence Deletion / Q000235:genetics; D013234:Stem Cells / Q000378:metabolism*; D014157:Transcription Factors / Q000235:genetics / Q000378:metabolism / Q000502:physiology*; D014158:Transcription, Genetic
chemical_list: D004268:DNA-Binding Proteins; D024002:HMGB Proteins; D018398:Homeodomain Proteins; C486780:NANOG protein, human; D000071317:Nanog Homeobox Protein; C475540:Nanog protein, mouse; D009687:Nuclear Proteins; D050814:Octamer Transcription Factor-3; C494173:POU5F1 protein, human; C494174:Pou5f1 protein, mouse; C527782:SOX2 protein, human; D055748:SOXB1 Transcription Factors; C527783:Sox2 protein, mouse; D014157:Transcription Factors; C485184:enhanced green fluorescent protein; D049452:Green Fluorescent Proteins; D008156:Luciferases
references.citation: Mol Cell Biol. 1998 Apr;18(4):1866-78
references.pmid: 9528758
references.citation: EMBO J. 1997 Jun 2;16(11):3219-32
references.pmid: 9214638
references.citation: Cell. 1998 Oct 30;95(3):379-91
references.pmid: 9814708
references.citation: Mol Cell Biol. 1999 Aug;19(8):5453-65
references.pmid: 10409735
references.citation: Mech Dev. 2005 Jan;122(1):67-79
references.pmid: 15582778
references.citation: Gene. 2003 Dec 24;323:163-72
references.pmid: 14659890
references.citation: FEBS Lett. 1999 Nov 12;461(1-2):125-9
references.pmid: 10561509
references.citation: Nat Genet. 2000 Apr;24(4):372-6
references.pmid: 10742100
references.citation: Gene. 2000 Apr 4;246(1-2):383-93
references.pmid: 10767561
references.citation: Mol Reprod Dev. 2000 Sep;57(1):4-15
references.pmid: 10954851
references.citation: Mol Cell Biol. 2001 Jul;21(13):4330-6
references.pmid: 11390661
references.citation: Cell Struct Funct. 2001 Jun;26(3):149-60
references.pmid: 11565807
references.citation: Nature. 2002 Jul 18;418(6895):293-300
references.pmid: 12124616
references.citation: Nucleic Acids Res. 2002 Jul 15;30(14):3202-13
references.pmid: 12136102
references.citation: Cell. 2002 Oct 18;111(2):185-96
references.pmid: 12408863
references.citation: Genes Dev. 2003 Jan 1;17(1):126-40
references.pmid: 12514105
references.citation: Gene Expr Patterns. 2003 Mar;3(1):99-103
references.pmid: 12609610
references.citation: Biochem Biophys Res Commun. 2003 Mar 14;302(3):581-6
references.pmid: 12615074
references.citation: Mol Cell Biol. 2003 Apr;23(8):2699-708
references.pmid: 12665572
references.citation: Stem Cells. 2003;21(3):257-65
references.pmid: 12743320
references.citation: Cell. 2003 May 30;113(5):631-42
references.pmid: 12787504
references.citation: Cell. 2003 May 30;113(5):643-55
references.pmid: 12787505
references.citation: Development. 2003 Sep;130(18):4235-48
references.pmid: 12900441
references.citation: Cell. 2003 Oct 31;115(3):281-92
references.pmid: 14636556
references.citation: Dev Dyn. 2004 May;230(1):187-98
references.pmid: 15108323
references.citation: Mol Cell Biol. 2004 Jul;24(13):5710-20
references.pmid: 15216876
references.citation: J Embryol Exp Morphol. 1980 Aug;58:289-302
references.pmid: 7441158
references.citation: Nature. 1988 Dec 15;336(6200):684-7
references.pmid: 3143916
references.citation: Nature. 1988 Dec 15;336(6200):688-90
references.pmid: 3143917
references.citation: EMBO J. 1989 Sep;8(9):2543-50
references.pmid: 2573523
references.citation: EMBO J. 1990 Nov;9(11):3723-32
references.pmid: 1976514
references.citation: Development. 1992 Mar;114(3):755-68
references.pmid: 1618140
references.citation: Nature. 1992 Sep 3;359(6390):76-9
references.pmid: 1522892
references.citation: Nature. 1993 Mar 4;362(6415):79-83
references.pmid: 7680434
references.citation: Science. 1993 Jun 18;260(5115):1805-8
references.pmid: 8390097
references.citation: Mol Cell Biol. 1994 Dec;14(12):7758-69
references.pmid: 7969117
references.citation: Development. 1995 May;121(5):1283-99
references.pmid: 7789261
references.citation: Genes Dev. 1995 Sep 1;9(17):2105-16
references.pmid: 7657163
references.citation: Genes Dev. 1995 Nov 1;9(21):2635-45
references.pmid: 7590241
references.citation: Development. 1996 Mar;122(3):881-94
references.pmid: 8631266
references.citation: Proc Natl Acad Sci U S A. 1997 Apr 15;94(8):3801-4
references.pmid: 9108058
references.citation: Science. 1998 Nov 6;282(5391):1145-7
references.pmid: 9804556