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An Oct4-Centered Protein Interaction Network in Embryonic Stem Cells

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...

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Autores principales: van den Berg, Debbie L.C., Snoek, Tim, Mullin, Nick P., Yates, Adam, Bezstarosti, Karel, Demmers, Jeroen, Chambers, Ian, Poot, Raymond A.
Formato: Texto
Lenguaje:English
Publicado: Cell Press 2010
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2860243/
https://www.ncbi.nlm.nih.gov/pubmed/20362541
http://dx.doi.org/10.1016/j.stem.2010.02.014
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author van den Berg, Debbie L.C.
Snoek, Tim
Mullin, Nick P.
Yates, Adam
Bezstarosti, Karel
Demmers, Jeroen
Chambers, Ian
Poot, Raymond A.
author_facet van den Berg, Debbie L.C.
Snoek, Tim
Mullin, Nick P.
Yates, Adam
Bezstarosti, Karel
Demmers, Jeroen
Chambers, Ian
Poot, Raymond A.
author_sort van den Berg, Debbie L.C.
collection PubMed
description 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.
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spelling pubmed-28602432010-05-05 An Oct4-Centered Protein Interaction Network in Embryonic Stem Cells van den Berg, Debbie L.C. Snoek, Tim Mullin, Nick P. Yates, Adam Bezstarosti, Karel Demmers, Jeroen Chambers, Ian Poot, Raymond A. Cell Stem Cell Resource 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. Cell Press 2010-04-02 /pmc/articles/PMC2860243/ /pubmed/20362541 http://dx.doi.org/10.1016/j.stem.2010.02.014 Text en © 2010 ELL & Excerpta Medica. https://creativecommons.org/licenses/by/3.0/ Open Access under CC BY 3.0 (https://creativecommons.org/licenses/by/3.0/) license
spellingShingle Resource
van den Berg, Debbie L.C.
Snoek, Tim
Mullin, Nick P.
Yates, Adam
Bezstarosti, Karel
Demmers, Jeroen
Chambers, Ian
Poot, Raymond A.
An Oct4-Centered Protein Interaction Network in Embryonic Stem Cells
title An Oct4-Centered Protein Interaction Network in Embryonic Stem Cells
title_full An Oct4-Centered Protein Interaction Network in Embryonic Stem Cells
title_fullStr An Oct4-Centered Protein Interaction Network in Embryonic Stem Cells
title_full_unstemmed An Oct4-Centered Protein Interaction Network in Embryonic Stem Cells
title_short An Oct4-Centered Protein Interaction Network in Embryonic Stem Cells
title_sort oct4-centered protein interaction network in embryonic stem cells
topic Resource
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2860243/
https://www.ncbi.nlm.nih.gov/pubmed/20362541
http://dx.doi.org/10.1016/j.stem.2010.02.014
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