Site-specific Disruption of the Oct4/Sox2 Protein Interaction Reveals Coordinated Mesendodermal Differentiation and the Epithelial-Mesenchymal Transition

Pan, Xiao; Cang, Xiaohui; Dan, Songsong; Li, Jingchao; Cheng, Jie; Kang, Bo; Duan, Xiaotao; Shen, Binghui; Wang, Yingjie

doi:10.1074/jbc.m116.745414

Cited by 32 publications

(23 citation statements)

References 45 publications

(66 reference statements)

Supporting

Mentioning

Contrasting

Unclassified

Order By: Relevance

“…The distributions and median expression levels of these proteins were similar to those of wild‐type E14 ES cells (Fig D). We also found the half‐lives of both OCT4‐HALO (7.8 ± 1.3 h) and SOX2‐SNAP (8.1 ± 1.0 h) to be close to published half‐life values for these proteins (Fang et al , ; Pan et al , ; Liu et al , ) (Fig E and Table EV1), and the average cell cycle length of SBROS and wild‐type E14 ES cells were similar (Fig F and Table EV2). mRNA levels of pluripotency markers of SBROS cells were mostly unaltered (Fig EV1E and Table EV3).…”

Section: Resultssupporting

confidence: 85%

Endogenous fluctuations of OCT 4 and SOX 2 bias pluripotent cell fate decisions

Strebinger

Deluz

Friman

et al. 2019

Molecular Systems Biology

View full text Add to dashboard Cite

SOX2 and OCT4 are pioneer transcription factors playing a key role in embryonic stem (ES) cell self‐renewal and differentiation. How temporal fluctuations in their expression levels bias lineage commitment is unknown. Here, we generated knock‐in reporter fusion ES cell lines allowing to monitor endogenous SOX2 and OCT4 protein fluctuations in living cells and to determine their impact on mesendodermal and neuroectodermal commitment. We found that small differences in SOX2 and OCT4 levels impact cell fate commitment in G1 but not in S phase. Elevated SOX2 levels modestly increased neuroectodermal commitment and decreased mesendodermal commitment upon directed differentiation. In contrast, elevated OCT4 levels strongly biased ES cells towards both neuroectodermal and mesendodermal fates in undirected differentiation. Using ATAC‐seq on ES cells gated for different endogenous SOX2 and OCT4 levels, we found that high OCT4 levels increased chromatin accessibility at differentiation‐associated enhancers. This suggests that small endogenous fluctuations of pioneer transcription factors can bias cell fate decisions by concentration‐dependent priming of differentiation‐associated enhancers.

show abstract

Section: Resultssupporting

confidence: 85%

Endogenous fluctuations of OCT 4 and SOX 2 bias pluripotent cell fate decisions

Strebinger

Deluz

Friman

et al. 2019

Molecular Systems Biology

View full text Add to dashboard Cite

show abstract

“…The distributions and median expression levels of these proteins were similar to those of wild type E14 ES cells ( Fig.1d). We also found the half-lives of both OCT4-HALO (7.8 ±1.3 h) and SOX2-SNAP (8.1 ±1.0 h) to be close to published half-life values for these proteins (Pan et al, 2016;Fang et al, 2014;Liu et al, 2017a) (Fig.1e), and the average cell cycle length of SBROS and wild type E14 ES cells were similar (Fig.1f). mRNA levels of pluripotency markers of SBROS cells were mostly unaltered ( Supplementary Fig.1e).…”

Section: Generation Of a Sox2-snap / Oct4-halo Knock-in Es Cell Linesupporting

confidence: 74%

Endogenous fluctuations of OCT4 and SOX2 bias pluripotent cell fate decisions

Strebinger

Friman

Deluz

et al. 2018

Preprint

View full text Add to dashboard Cite

SOX2 and OCT4 are pioneer transcription factors playing a key role in embryonic stem (ES) cell self-renewal and differentiation. However, how temporal fluctuations in their expression levels bias lineage commitment is unknown. Here we generated knock-in reporter fusion ES cell lines allowing to monitor endogenous SOX2 and OCT4 protein fluctuations in living cells and to determine their impact on mesendodermal and neuroectodermal commitment. We found that small differences in SOX2 and OCT4 levels impact cell fate commitment in G1 but not in S phase. Elevated SOX2 levels modestly increased neuroectodermal commitment and decreased mesendodermal commitment upon directed differentiation. In contrast, elevated OCT4 levels strongly biased ES cell towards both neuroectodermal and mesendodermal fates. Using ATAC-seq on ES cells gated for different endogenous SOX2 and OCT4 levels, we found that high OCT4 levels increased chromatin accessibility at differentiation-associated enhancers. This suggests that small endogenous fluctuations of pioneer transcription factors can bias cell fate decisions by concentration-dependent priming of differentiation-associated enhancers. KeywordsEmbryonic stem cells/Endogenous fluctuations/SOX2/OCT4/Differentiation

show abstract

“…2D). Since OCT4 and SOX2 can form a complex, bind to downstream target genes, and maintain the pluripotency32, the multiple transduction of OSKM factors were applied. Results showed that OCT4-SOX2 complex as well as OSK and OSM combinants could significantly reduce EpCAM promoter activity, but combinations of OCT4-KLF4, OCT4-c-MYC, SOX2-KLF4, SOX2-c-MYC, and KLF4-c-MYC did not influence EpCAM activity.…”

Section: Resultsmentioning

confidence: 99%

EpCAM Intracellular Domain Promotes Porcine Cell Reprogramming by Upregulation of Pluripotent Gene Expression via Beta-catenin Signaling

Wang

2017

Sci Rep

View full text Add to dashboard Cite

Previous study showed that expression of epithelial cell adhesion molecule (EpCAM) was significantly upregulated in porcine induced pluripotent stem cells (piPSCs). However, the regulatory mechanism and the downstream target genes of EpCAM were not well investigated. In this study, we found that EpCAM was undetectable in fibroblasts, but highly expressed in piPSCs. Promoter of EpCAM was upregulated by zygotic activated factors LIN28, and ESRRB, but repressed by maternal factors OCT4 and SOX2. Knocking down EpCAM by shRNA significantly reduced the pluripotent gene expression. Conversely, overexpression of EpCAM significantly increased the number of alkaline phosphatase positive colonies and elevated the expression of endogenous pluripotent genes. As a key surface-to-nucleus factor, EpCAM releases its intercellular domain (EpICD) by a two-step proteolytic processing sequentially. Blocking the proteolytic processing by inhibitors TAPI-1 and DAPT could reduce the intracellular level of EpICD and lower expressions of OCT4, SOX2, LIN28, and ESRRB. We noticed that increasing intracellular EpICD only was unable to improve activity of EpCAM targeted genes, but by blocking GSK-3 signaling and stabilizing beta-catenin signaling, EpICD could then significantly stimulate the promoter activity. These results showed that EpCAM intracellular domain required beta-catenin signaling to enhance porcine cell reprogramming.

show abstract

Site-specific Disruption of the Oct4/Sox2 Protein Interaction Reveals Coordinated Mesendodermal Differentiation and the Epithelial-Mesenchymal Transition

Cited by 32 publications

References 45 publications

Endogenous fluctuations of OCT 4 and SOX 2 bias pluripotent cell fate decisions

Endogenous fluctuations of OCT 4 and SOX 2 bias pluripotent cell fate decisions

Endogenous fluctuations of OCT4 and SOX2 bias pluripotent cell fate decisions

EpCAM Intracellular Domain Promotes Porcine Cell Reprogramming by Upregulation of Pluripotent Gene Expression via Beta-catenin Signaling

Contact Info

Product

Resources

About