A conserved H3K14ub-driven H3K9me3 for chromatin compartmentalization

To delineate the relationship between G2E3-catalyzed H3K14ub and H3K9me3 in genome-wide, we performed chromatin immunoprecipitation followed by high-throughput sequencing (ChIP-seq) for H3K14ub, H3K9me3 and H3K4me3 in Parental (sgG2E3) control HeLa cells and G2E3 KO-10 HeLa cells. Our data demonstrated that G2E3-catalyzed H3K14ub potentiates H3K9 trimethylation (H3K9me3) by SUV39H and is specifically required for SUV39H compartmentalization and H3K9me3 in pericentromeric heterochromatin. Loss ofG2E3 severely impairs not only pericentromeric heterochromatin organization, but also results in aberrant accumulation of SUV39H and H3K9me3 in numerous euchromatin regions and a widespread transcriptional repression (combined with RNA-seq data). Overall design: Chromatin immunoprecipitation DNA-sequencing (ChIP-seq) for histone modifications H3K4me3, H3K9me3 and H3K14ub in Parental (sgG2E3)control HeLa cells and G2E3 knockout HeLa cells. sgG2E3 control HeLa cells is a sgRNA stable cell line that has not been cut by Cas9 protein. In our paper, it is marked as Parental control HeLa cells.