H2BK5ac is required for the establishment of inflammatory potential in mouse embryonic stem cells following differentiation.
Embryonic stem cells (ESCs) are refractory to inflammatory stimuli but acquire inflammatory competence upon differentiation, yet the underlying mechanisms remain unclear. Here, we report that H2BK5ac regulates the acquisition of inflammatory potential in mouse ESCs during differentiation. H2BK5ac levels increased at the regulatory regions of inflammatory genes upon differentiation, and this modification was required for their transcriptional upregulation. An acetylation-deficient H2BK5A mutant impaired the upregulation of inflammatory genes, whereas an acetylation-mimicking H2BK5Q mutant had the opposite effect. We identified p300 and HDAC3 as the acetyltransferase and deacetylase, respectively, that control H2BK5ac: p300 protein levels increased upon differentiation and its depletion reduced H2BK5ac levels, whereas loss of HDAC3 increased H2BK5ac levels despite its unchanged protein levels. H2BK5Q overexpression in mouse embryos increased embryonic mortality and inflammatory cytokine secretion, consistent with its pro-inflammatory effect. Thus, our study reveals H2BK5ac as a regulator that establishes inflammatory potential in mouse ESCs during differentiation.