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NANOG alone induces germ cells in primed epiblast in vitro by activation of enhancers.


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Authors

Murakami, Kazuhiro 
Günesdogan, Ufuk 
Zylicz, Jan J 
Tang, Walfred WC 
Sengupta, Roopsha 

Abstract

Nanog, a core pluripotency factor in the inner cell mass of blastocysts, is also expressed in unipotent primordial germ cells (PGCs) in mice, where its precise role is yet unclear. We investigated this in an in vitro model, in which naive pluripotent embryonic stem (ES) cells cultured in basic fibroblast growth factor (bFGF) and activin A develop as epiblast-like cells (EpiLCs) and gain competence for a PGC-like fate. Consequently, bone morphogenetic protein 4 (BMP4), or ectopic expression of key germline transcription factors Prdm1, Prdm14 and Tfap2c, directly induce PGC-like cells (PGCLCs) in EpiLCs, but not in ES cells. Here we report an unexpected discovery that Nanog alone can induce PGCLCs in EpiLCs, independently of BMP4. We propose that after the dissolution of the naive ES-cell pluripotency network during establishment of EpiLCs, the epigenome is reset for cell fate determination. Indeed, we found genome-wide changes in NANOG-binding patterns between ES cells and EpiLCs, indicating epigenetic resetting of regulatory elements. Accordingly, we show that NANOG can bind and activate enhancers of Prdm1 and Prdm14 in EpiLCs in vitro; BLIMP1 (encoded by Prdm1) then directly induces Tfap2c. Furthermore, while SOX2 and NANOG promote the pluripotent state in ES cells, they show contrasting roles in EpiLCs, as Sox2 specifically represses PGCLC induction by Nanog. This study demonstrates a broadly applicable mechanistic principle for how cells acquire competence for cell fate determination, resulting in the context-dependent roles of key transcription factors during development.

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Keywords

Activins, Animals, Bone Morphogenetic Protein 4, Cell Differentiation, Chromatin, DNA-Binding Proteins, Enhancer Elements, Genetic, Epigenesis, Genetic, Female, Fibroblast Growth Factor 2, Gene Expression Regulation, Developmental, Genome, Germ Cells, Germ Layers, Homeodomain Proteins, Male, Mice, Mouse Embryonic Stem Cells, Nanog Homeobox Protein, Pluripotent Stem Cells, Positive Regulatory Domain I-Binding Factor 1, Protein Binding, RNA-Binding Proteins, SOXB1 Transcription Factors, Transcription Factor AP-2, Transcription Factors

Journal Title

Nature

Conference Name

Journal ISSN

0028-0836
1476-4687

Volume Title

529

Publisher

Springer Science and Business Media LLC
Sponsorship
Medical Research Council (MC_PC_12009)
Wellcome Trust (092096/Z/10/Z)
Wellcome Trust (096738/Z/11/Z)
Cancer Research Uk (None)