Histone H3.3 ensures cell proliferation and genomic stability during myeloid cell development [RNA-seq]

Variant histone H3.3 is thought to be critical for survival of many cells, since it is deposited in expressed genes, a feature different from core histones. For example, H3.3 deletion leads to embryonic lethality in mice. However, requirement of H3.3 in later stage of development has remained unclear. The aim of this work was to elucidate the role of H3.3 for development of myeloid lineage, important for innate immunity. We conditionally knocked out (cKO) the H3.3 genes in myeloid progenitor cells differentiating into bone marrow derived macrophages (BMDMs). Progenitor cells lacking H3.3 were defective in replication, suffered from extensive DNA damage, and underwent apoptosis. Surviving H3.3cKO cells expressed many interferon stimulated genes (ISGs) throughout differentiation. Further, H3.3cKO BMDMs possessed chromatin accessible sites, and histone posttranslational modifications consistent with the gene expression profiles, Accordingly, H3.3cKO BMDMs retained general nucleosomal structure genome wide. In summary, H3.3 is required for proliferation of myeloid progenitor cells, but is in large part dispensable for differentiation of BMDMs. RNA seq of WT and H3.3 cKO cells on day3, 5 and day7 of M-CSF induced differentiation. RNA seq of WT and H3.3 cKO BMDM on day8 of M-CSF induced differentiation under untreated and IFNγ treated (12 hrs) conditions RNA seq of WT and H3.3 cKO peritoneal macrophages. RNA seq of WT and cKO BMDMs under no antibody, control antibody and IFNAR1 antibody conditions. RNA seq of WT, H3.3cKO and H3.3cKO:Sting KO BMDMs. RNA seq of WT, H3.3cKO and H3.3cKO:Mavs KO BMDMs. RNA seq of WT, H3.3cKO and H3.3cKO:Irf7 KO BMDMs.