Complementary Activity of ETV5, RBPJ, and TCF3 Drives Formative Transition from Naive Pluripotency

Cell Stem Cell. 2019 May 2;24(5):785-801.e7. doi: 10.1016/j.stem.2019.03.017. Epub 2019 Apr 25.


The gene regulatory network (GRN) of naive mouse embryonic stem cells (ESCs) must be reconfigured to enable lineage commitment. TCF3 sanctions rewiring by suppressing components of the ESC transcription factor circuitry. However, TCF3 depletion only delays and does not prevent transition to formative pluripotency. Here, we delineate additional contributions of the ETS-family transcription factor ETV5 and the repressor RBPJ. In response to ERK signaling, ETV5 switches activity from supporting self-renewal and undergoes genome relocation linked to commissioning of enhancers activated in formative epiblast. Independent upregulation of RBPJ prevents re-expression of potent naive factors, TBX3 and NANOG, to secure exit from the naive state. Triple deletion of Etv5, Rbpj, and Tcf3 disables ESCs, such that they remain largely undifferentiated and locked in self-renewal, even in the presence of differentiation stimuli. Thus, genetic elimination of three complementary drivers of network transition stalls developmental progression, emulating environmental insulation by small-molecule inhibitors.

Keywords: ETS factors; RBPJ; commitment; differentiation; embryonic stem cell; epiblast; gene regulatory network; pluripotency; self-renewal.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Basic Helix-Loop-Helix Transcription Factors / genetics
  • Basic Helix-Loop-Helix Transcription Factors / metabolism*
  • Cell Differentiation
  • Cell Line
  • Cell Lineage
  • Cell Self Renewal
  • DNA-Binding Proteins / genetics
  • DNA-Binding Proteins / metabolism*
  • Gene Knockout Techniques
  • Gene Regulatory Networks
  • Immunoglobulin J Recombination Signal Sequence-Binding Protein / genetics
  • Immunoglobulin J Recombination Signal Sequence-Binding Protein / metabolism*
  • Mice
  • Mice, Inbred C57BL
  • Mice, Inbred CBA
  • Nanog Homeobox Protein / genetics
  • Nanog Homeobox Protein / metabolism
  • Neurons / physiology*
  • Pluripotent Stem Cells / physiology*
  • RNA, Small Interfering / genetics
  • T-Box Domain Proteins / genetics
  • T-Box Domain Proteins / metabolism
  • Transcription Factors / genetics
  • Transcription Factors / metabolism*


  • Basic Helix-Loop-Helix Transcription Factors
  • DNA-Binding Proteins
  • Etv5 protein, mouse
  • Immunoglobulin J Recombination Signal Sequence-Binding Protein
  • Nanog Homeobox Protein
  • Nanog protein, mouse
  • RNA, Small Interfering
  • Rbpj protein, mouse
  • T-Box Domain Proteins
  • Tbx3 protein, mouse
  • Tcf3 protein, mouse
  • Transcription Factors