H2A.Z facilitates Sox2-nucleosome interaction by promoting DNA and histone H3 tail mobility.

Epigenetic regulation of eukaryotic chromatin structure and function can be modulated by histone variants and post-translational modifications. The conserved variant H2A.Z has been functionally linked to pioneer factors Sox2 and Oct4 that open chromatin and initiate cell fate-specific expression programs. However, the molecular basis for their interaction remains unknown. Using biochemistry, nuclear magnetic resonance (NMR) spectroscopy and molecular dynamics (MD) simulations, we examine the role of H2A.Z nucleosome dynamics in pioneer factor binding. We find that H2A.Z facilitates Sox2 and Oct4 binding at distinct locations in 601 nucleosomes. We further link this to increased DNA accessibility and perturbed dynamics of the H3 N-terminal tail, which we show competes with Sox2 for DNA binding. Our simulations validate a coupling between H2A.Z-mediated DNA unwrapping and altered H3 N-tail conformations with fewer contacts to DNA and the H2A.Z C- terminal tail. This destabilizing effect of H2A.Z is DNA sequence dependent and enhanced with the less stable Lin28B nucleosome. Collectively, our findings suggest that H2A.Z promotes pioneer factor binding by increasing access to DNA and reducing competition with H3 tails. This could have broader implications for how epigenetic marks or oncogenic mutations tune pioneer factor engagement with chromatin and thus affect its structure and recognition.