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Cell Research (2006)16: 319-322
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Functional analysis of two Sp1/Sp3 binding sites in murine Nanog gene promoter

Da Yong Wu¹, Zhen Yao¹

¹1Laboratory 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

Correspondence: Da Yong Wu Tel: 86-21-54921367; E-mail: dywu@sibs.ac.cn Received 14 Aug 2005; revised 26 Nov 2005; accepted 20 Dec 2005; published online 16 March 2006

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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.

Cell Research (2006) 16:319-322. doi:10.1038/sj.cr.7310040; published online 16 March 2006

Keywords: Nanog, promoter, Sp1, Sp3