H3K9me3 and MORC2 ChIP-seq profiling of wild-type and MORC2-knock-out HEK293T cells

We report an in vitro reconstitution of full-length MORC2, the most commonly mutated MORC member, linked to various cancers and neurological disorders. MORC2 possesses multiple DNA binding sites that undergo structural rearrangement upon DNA binding. MORC2 locks onto the DNA using its C-terminal domain (CTD) and acts as a sliding clamp. A conserved phosphate-interacting motif within the CTD was found to regulate ATP hydrolysis and cooperative DNA binding. Importantly, MORC2 mediates chromatin remodelling via ATP hydrolysis-dependent DNA compaction, regulated by the phosphorylation state of its CTD. Overall design: To identify the preferred genomic regions of MORC2 binding, we performed chromatin immunoprecipitation sequencing (ChIP-seq) on the HEK293T cell line utilising antibodies specific for MORC2 and H3K9me3, a mark of constitutive heterochromatin. Negative control IgG ChIP-seq experiments were performed. All sequencing experiments were run in duplicate on wild-type and MORC2-knock-out HEK293T cell lines.