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KMT2C and KMT2D amplify GRHL2-driven enhancer activation.

Ryan M Boileau1,2,3,4,5, Kevin X Chen1,2,3, Robert Blelloch1,2,3

  • 1The Eli and Edythe Broad Center of Regeneration Medicine and Stem Cell Research, Center for Reproductive Sciences, University of California San Francisco, San Francisco, USA.

Biorxiv : the Preprint Server for Biology
|March 27, 2026
PubMed
Summary
This summary is machine-generated.

We developed a new system to study enhancer activation in embryonic stem cells (ESCs). This system reveals that histone modifiers KMT2C/D amplify the effects of the pioneer transcription factor GRHL2, crucial for cell fate specification.

Keywords:
EnhancerGRHL2H3K27acH3K4me1KMT2CKMT2DMLL3MLL4P300pioneer

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Area of Science:

  • Developmental Biology
  • Epigenetics
  • Gene Regulation

Background:

  • Enhancer activation is critical for cell fate specification and cell type-specific gene expression.
  • Studying enhancer biology during cell differentiation is challenging due to asynchronous gene regulatory changes.

Purpose of the Study:

  • To develop a tamoxifen-inducible system for acute enhancer activation in embryonic stem cells (ESCs).
  • To investigate the functional relationship between the pioneer transcription factor GRHL2 and histone modifiers KMT2C/D.

Main Methods:

  • Development of a tamoxifen-gated system for acute enhancer activation in ESCs using GRHL2.
  • Analysis of histone modifications (H3K4me1/2, H3K27ac), co-factor recruitment (P300), and transcriptional activation.
  • Investigation during naive to formative ESC differentiation.

Main Results:

  • GRHL2 binds target sites independently of KMT2C/D.
  • KMT2C/D are required for robust H3K4me1/2, P300, and H3K27ac deposition and transcriptional activation at GRHL2 sites.
  • A basal level of enhancer activation and transcription occurs even without KMT2C/D.
  • GRHL2-mediated enhancer remodeling during ESC differentiation is dependent on KMT2C/D.

Conclusions:

  • KMT2C/D act as essential co-activators, amplifying GRHL2-driven enhancer activation in ESCs.
  • The developed inducible system enables dissection of the kinetics and enzymatic dependencies of pioneer transcription factor-mediated enhancer remodeling.