Dynamical reorganization of the pluripotency transcription factors Oct4 and Sox2 during early differentiation of embryonic stem cells

Abstract

Pluripotency maintenance requires transcription factors (TFs) that induce genes necessary to preserve the undifferentiated state and repress others involved in differentiation. Recent observations support that the heterogeneous distribution of TFs in the nucleus impacts on gene expression. Thus, it is essential to explore how TFs dynamically organize to fully understand their role in transcription regulation. Here, we examine the distribution of pluripotency TFs Oct4 and Sox2 in the nucleus of embryonic stem (ES) cells and inquire whether their organization changes during early differentiation stages preceding their downregulation. Using ES cells expressing Oct4-YPet or Sox2-YPet, we show that Oct4 and Sox2 partition between nucleoplasm and a few chromatin-dense foci which restructure after inducing differentiation by 2i/LIF withdrawal. Fluorescence correlation spectroscopy showed distinct changes in Oct4 and Sox2 dynamics after differentiation induction. Specifically, we detected an impairment of Oct4-chromatin interactions whereas Sox2 only showed slight variations in its shortlived, and probably more unspecific, interactions with chromatin. Our results reveal that differentiation cues trigger early changes of Oct4 and Sox2 nuclear distributions that also include modifications in TF-chromatin interactions. This dynamical reorganization precedes Oct4 and Sox2 downregulation and may contribute to modulate their function at early differentiation stages.

Ubiquitin-Specific Protease 3 Deubiquitinates and Stabilizes Oct4 Protein in Human Embryonic Stem Cells

“…Interestingly, Western blot analysis of EB lysates at various time points revealed a transient reduction in USP3 expression at day 2 and recovery from day 3 (Figure 5B). However, the expression of Oct4 gradually decreased during differentiation, consistent with a previous report [33] (Figure 5B).…”

Neural precursors cells expanded in a 3D micro-engineered niche present enhanced therapeutic efficacy in vivo

“…Indeed, all these genes converge to the activation of a core transcriptional network involving SOX2, NANOG and OCT4, also found up-regulated in NPCs expanded inside the Nichoid (Figure 2E, Figure 2F). These factors are expressed at high levels in embryonic stem cells [57]. Moreover, they regulate the expression of other genes during development and are found at high levels in pluripotent stem cells [58][59][60].…”

An Esrrb and Nanog Cell Fate Regulatory Module Controlled by Feed Forward Loop Interactions

“…Understanding how embryonic and induced pluripotent stem cells (ESCs and iPSCs, respectively) regulate cell fate decisions is crucial to realizing their biomedical potential ( Liu et al, 2020 ). The pluripotent state is maintained by intrinsic and extrinsic signals that converge on a network of core transcription factors (TFs) including Oct4, Sox2, and Nanog among others ( Boiani and Schöler, 2005 ; Ivanova et al, 2006 ; Wang et al, 2006 ; Chen et al, 2008 ; Papatsenko et al, 2015 ; Verneri et al, 2020 ). These factors are interconnected via transcriptional and protein-protein interactions ( Wang et al, 2006 ; Pardo et al, 2010 ; van den Berg et al, 2010 ).…”

“…From this point of view, network regulatory functions have clear multidimensionality. While previously many studies were focused on one or two dimensions (e.g., mRNA expression and/or TF binding) at a single time-point, now more studies are starting to address the dynamics of multidimensionality of regulatory networks as they process biological information during cell fate transitions ( Lu et al, 2009 ; Verneri et al, 2020 ). The goal of the current study is to elucidate global dynamic changes across multiple regulatory dimensions promoted by the depletion of Estrogen related receptor beta (Esrrb), a major core pluripotency TF.…”