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Pluripotency governed by Sox2 via regulation of Oct3/4 expression in mouse embryonic stem cells

Nature Cell Biology volume 9, pages 625–635 (2007)Cite this article

Abstract

The pluripotency of embryonic stem (ES) cells is thought to be maintained by a few key transcription factors, including Oct3/4 and Sox2. The function of Oct3/4 in ES cells has been extensively characterized, but that of Sox2 has yet to be determined. Sox2 can act synergistically with Oct3/4 in vitro to activate Oct–Sox enhancers, which regulate the expression of pluripotent stem cell-specific genes, including Nanog, Oct3/4 and Sox2 itself. These findings suggest that Sox2 is required by ES cells for its Oct–Sox enhancer activity. Using inducible Sox2-null mouse ES cells, we show that Sox2 is dispensable for the activation of these Oct–Sox enhancers. In contrast, we demonstrate that Sox2 is necessary for regulating multiple transcription factors that affect Oct3/4 expression and that the forced expression of Oct3/4 rescues the pluripotency of Sox2-null ES cells. These results indicate that the essential function of Sox2 is to stabilize ES cells in a pluripotent state by maintaining the requisite level of Oct3/4 expression.

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Figure 1: Sox2-null ES cells differentiate into trophoectoderm-like cells.
Figure 2: Sox2-null ES cells express trophoectoderm marker genes.
Figure 3: Oct–Sox enhancer dependent genes are slowly downregulated on Sox2-null induction.
Figure 4: Redundancy of Sox factors on Oct–Sox enhancers.
Figure 5: Sox2-null can be rescued by the forced expression of Oct3/4.
Figure 6: Characterization of rescued Sox2-null cells.
Figure 7: Schematic representation of a model of a transcriptional network governed by Oct3/4 and Sox factors.

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Acknowledgements

We thank H. Bujard (University of Heidelberg) for providing the plasmid pUHD-tetR/VP16(FF); I. R. Lemischka (Princeton University) for the Sox2 knockdown cell lines; S. Miyagi for RNA samples from E11.5 embryos; and Y. Piao for microarray hybridization. This work was supported in part by grant-in-aids for Scientific Research from the Ministry of Education, Culture, Sports, Science and Technology of Japan (to S.M.) and by a RIKEN grant and grants for the 21st Century COE Program, from the Ministry of Education, Culture, Sports, Science, and Technology of Japan and the Leading Project (to H.N.). H.N. also received funding from the CREST program of the Japan Science and Technology Agency. The work was also supported in part by the Intramural Research Program of the National Institute on Aging, National Institutes of Health.

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Authors and Affiliations

  1. Laboratory for Pluripotent Cell Studies, RIKEN Center for Developmental Biology, Minatojima-minamimachi 2-2-3, Chu-o-ku, Kobe, Hyogo, 650-0047, Japan

    Shinji Masui, Yuhki Nakatake, Yayoi Toyooka, Daisuke Shimosato, Rika Yagi, Kazue Takahashi & Hitoshi Niwa

  2. CREST (Core Research for Evolutional Science and Technology), Japan Science and Technology Agency, Honcho 4-1-8, Kawaguchi, Saitama, 332-0012, Japan

    Shinji Masui & Hitoshi Niwa

  3. Division of Molecular Biology and Cell Engineering, Department of Regenerative Medicine, Research Institute, International Medical Center of Japan, 1-21-1 Toyama, Shinjuku-ku, Tokyo, 162-8655, Japan

    Shinji Masui

  4. Laboratory for Development and Regenerative Medicine, Kobe University Graduate School of Medicine, 7-5-1 Kusunokicho, Chu-o-ku, Kobe, Hyogo, 650-0017, Japan

    Daisuke Shimosato & Hitoshi Niwa

  5. Department of Regenerative Medicine, Research Institute, International Medical Center of Japan, 1-21-1 Toyama, Shinjuku-ku, Tokyo, 162-8655, Japan

    Hitoshi Okochi

  6. Division of Developmental Biology, Research Center for Genomic Medicine, Saitama Medical University, 1397-1 Yamane, Hidaka, Saitama, 350-1241, Japan

    Akihiko Okuda

  7. Developmental Genomics and Aging Section, Laboratory of Genetics, National Institute on Aging, National Institutes of Health, 333 Cassell Drive, Suite 3000, Baltimore, 21224-6820, MD, USA

    Ryo Matoba, Alexei A. Sharov & Minoru S. H. Ko

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Masui, S., Nakatake, Y., Toyooka, Y. et al. Pluripotency governed by Sox2 via regulation of Oct3/4 expression in mouse embryonic stem cells. Nat Cell Biol 9, 625–635 (2007). https://doi.org/10.1038/ncb1589

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