Functional analysis of two Sp1/Sp3 binding sites in murine Nanog gene promoter.
Nanog gene plays a key role in maintaining pluripotency of ES cells and early embryonic cells. A 5' flank sequence of the Nanog gene has been reported to be regulated differentially, and two regulatory elements within the Nanog promoter, namely Oct-4 and Sox-2 binding sites, have been identified to regulate the transcriptional activity of Nanog gene. In this report, we identified the role of two putative Sp1 binding sites located in the Nanog gene 5'-flanking region in regulation of murine Nanog gene transcription. Mutation studies showed that the two sites were essential for the Nanog promoter activity. Gel shift and supershift analysis showed that both sites specifically bind Sp1 and Sp3. Furthermore, overexpression of dominant-negative Sp1 or Sp3 mutants significantly inhibits Nanog promoter activity. These results suggest that the transcription factor Sp1 and Sp3 are important for Murine Nanog gene expression.
Laboratory of Molecular Cell Biology, Laboratory of Stem Cell Biology, Institute of Biochemistry and Cell Biology, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, 320 Yue Yang Road, Shanghai 200031, China. dywu@sibs.ac.cn
Da Yong
DY
Wu
Zhen
Z
Yao
D000818:Animals; D001483:Base Sequence; D001665:Binding Sites; D004268:DNA-Binding Proteins / Q000378:metabolism*; D024202:Electrophoretic Mobility Shift Assay; D018398:Homeodomain Proteins / Q000378:metabolism*; D051379:Mice; D008969:Molecular Sequence Data; D000071317:Nanog Homeobox Protein; D039904:Pluripotent Stem Cells / Q000502:physiology; D011401:Promoter Regions, Genetic; D016329:Sp1 Transcription Factor / Q000378:metabolism*; D051705:Sp3 Transcription Factor / Q000378:metabolism*
D004268:DNA-Binding Proteins; D018398:Homeodomain Proteins; D000071317:Nanog Homeobox Protein; C475540:Nanog protein, mouse; D016329:Sp1 Transcription Factor; C497122:Sp3 protein, mouse; D051705:Sp3 Transcription Factor