Octamer and Sox elements are required for transcriptional cis regulation of Nanog gene expression.
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.
Kuroda
Takao
T
Department of Development and Differentiation, Institute for Frontier Medical Sciences, Kyoto University, Shogoin, Sakyo-ku, Kyoto, Japan.
Tada
Masako
M
Kubota
Hiroshi
H
Kimura
Hironobu
H
Hatano
Shin-ya
SY
Suemori
Hirofumi
H
Nakatsuji
Norio
N
Tada
Takashi
T
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
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
Mol Cell Biol. 1998 Apr;18(4):1866-78
9528758
EMBO J. 1997 Jun 2;16(11):3219-32
9214638
Cell. 1998 Oct 30;95(3):379-91
9814708
Mol Cell Biol. 1999 Aug;19(8):5453-65
10409735
Mech Dev. 2005 Jan;122(1):67-79
15582778
Gene. 2003 Dec 24;323:163-72
14659890
FEBS Lett. 1999 Nov 12;461(1-2):125-9
10561509
Nat Genet. 2000 Apr;24(4):372-6
10742100
Gene. 2000 Apr 4;246(1-2):383-93
10767561
Mol Reprod Dev. 2000 Sep;57(1):4-15
10954851
Mol Cell Biol. 2001 Jul;21(13):4330-6
11390661
Cell Struct Funct. 2001 Jun;26(3):149-60
11565807
Nature. 2002 Jul 18;418(6895):293-300
12124616
Nucleic Acids Res. 2002 Jul 15;30(14):3202-13
12136102
Cell. 2002 Oct 18;111(2):185-96
12408863
Genes Dev. 2003 Jan 1;17(1):126-40
12514105
Gene Expr Patterns. 2003 Mar;3(1):99-103
12609610
Biochem Biophys Res Commun. 2003 Mar 14;302(3):581-6
12615074
Mol Cell Biol. 2003 Apr;23(8):2699-708
12665572
Stem Cells. 2003;21(3):257-65
12743320
Cell. 2003 May 30;113(5):631-42
12787504
Cell. 2003 May 30;113(5):643-55
12787505
Development. 2003 Sep;130(18):4235-48
12900441
Cell. 2003 Oct 31;115(3):281-92
14636556
Dev Dyn. 2004 May;230(1):187-98
15108323
Mol Cell Biol. 2004 Jul;24(13):5710-20
15216876
J Embryol Exp Morphol. 1980 Aug;58:289-302
7441158
Nature. 1988 Dec 15;336(6200):684-7
3143916
Nature. 1988 Dec 15;336(6200):688-90
3143917
EMBO J. 1989 Sep;8(9):2543-50
2573523
EMBO J. 1990 Nov;9(11):3723-32
1976514
Development. 1992 Mar;114(3):755-68
1618140
Nature. 1992 Sep 3;359(6390):76-9
1522892
Nature. 1993 Mar 4;362(6415):79-83
7680434
Science. 1993 Jun 18;260(5115):1805-8
8390097
Mol Cell Biol. 1994 Dec;14(12):7758-69
7969117
Development. 1995 May;121(5):1283-99
7789261
Genes Dev. 1995 Sep 1;9(17):2105-16
7657163
Genes Dev. 1995 Nov 1;9(21):2635-45
7590241
Development. 1996 Mar;122(3):881-94
8631266
Proc Natl Acad Sci U S A. 1997 Apr 15;94(8):3801-4
9108058
Science. 1998 Nov 6;282(5391):1145-7
9804556