Cell Stem Cell
Volume 6, Issue 4, 2 April 2010, Pages 369-381
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An Oct4-Centered Protein Interaction Network in Embryonic Stem Cells

https://doi.org/10.1016/j.stem.2010.02.014Get rights and content
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Summary

Transcription factors, such as Oct4, are critical for establishing and maintaining pluripotent cell identity. Whereas the genomic locations of several pluripotency transcription factors have been reported, the spectrum of their interaction partners is underexplored. Here, we use an improved affinity protocol to purify Oct4-interacting proteins from mouse embryonic stem cells (ESCs). Subsequent purification of Oct4 partners Sall4, Tcfcp2l1, Dax1, and Esrrb resulted in an Oct4 interactome of 166 proteins, including transcription factors and chromatin-modifying complexes with documented roles in self-renewal, but also many factors not previously associated with the ESC network. We find that Esrrb associated with the basal transcription machinery and also detect interactions between transcription factors and components of the TGF-β, Notch, and Wnt signaling pathways. Acute depletion of Oct4 reduced binding of Tcfcp2l1, Dax1, and Esrrb to several target genes. In conclusion, our purification protocol allowed us to bring greater definition to the circuitry controlling pluripotent cell identity.

Highlights

► An improved affinity method identified an Oct4 interactome of >160 proteins ► Iterative purification with Oct4 interactors helped define the network ► Network shows links to the basal transcription machinery and signaling pathways ► Transcription factor interactions may facilitate recruitment to the genome

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