Differential Histone Distribution Patterns in Induced Asymmetrically Dividing Mouse Embryonic Stem Cells

Highlights

• Old versus new H3 and H4 in dividing Wnt3a-induced mESCs do not overlap

• H4K20me2/3 is asymmetric on replicative chromatin fibers from Wnt3a-induced mESCs

• Old versus new H3 and H4 overlap in symmetrically dividing mESCs

• Old versus new H2A, H2B, and H3.3 overlap in dividing Wnt3a-induced mESCs

Summary

Wnt3a-coated beads can induce asymmetric divisions of mouse embryonic stem cells (mESCs), resulting in one self-renewed mESC and one differentiating epiblast stem cell. This provides an opportunity for studying histone inheritance pattern at a single-cell resolution in cell culture. Here, we report that mESCs with Wnt3a-bead induction display nonoverlapping preexisting (old) versus newly synthesized (new) histone H3 patterns, but mESCs without Wnt3a beads have largely overlapping patterns. Furthermore, H4K20me2/3, an old histone-enriched modification, displays a higher instance of asymmetric distribution on chromatin fibers from Wnt3a-induced mESCs than those from non-induced mESCs. These locally distinct distributions between old and new histones have both cellular specificity in Wnt3a-induced mESCs and molecular specificity for histones H3 and H4. Given that post-translational modifications at H3 and H4 carry the major histone modifications, our findings provide a mammalian cell culture system to study histone inheritance for maintaining stem cell fate and for resetting it during differentiation.